From bdc9648b877b8a8bf909cdc7b669189998c0edda Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 11 Apr 2019 11:35:32 +0100 Subject: [PATCH 001/149] Initial template commit --- .gitattributes | 1 + .github/CONTRIBUTING.md | 47 +++ .github/ISSUE_TEMPLATE/bug_report.md | 31 ++ .github/ISSUE_TEMPLATE/feature_request.md | 16 + .github/PULL_REQUEST_TEMPLATE.md | 15 + .github/markdownlint.yml | 9 + .gitignore | 7 + .travis.yml | 42 +++ CHANGELOG.md | 4 + CODE_OF_CONDUCT.md | 46 +++ Dockerfile | 7 + LICENSE | 21 ++ README.md | 30 ++ assets/email_template.html | 52 +++ assets/email_template.txt | 34 ++ assets/multiqc_config.yaml | 9 + assets/sendmail_template.txt | 36 ++ bin/markdown_to_html.r | 51 +++ bin/scrape_software_versions.py | 49 +++ conf/awsbatch.config | 18 + conf/base.config | 34 ++ conf/igenomes.config | 147 ++++++++ conf/test.config | 25 ++ docs/README.md | 12 + docs/output.md | 41 +++ docs/usage.md | 282 ++++++++++++++ environment.yml | 11 + main.nf | 428 ++++++++++++++++++++++ nextflow.config | 127 +++++++ 29 files changed, 1632 insertions(+) create mode 100644 .gitattributes create mode 100644 .github/CONTRIBUTING.md create mode 100644 .github/ISSUE_TEMPLATE/bug_report.md create mode 100644 .github/ISSUE_TEMPLATE/feature_request.md create mode 100644 .github/PULL_REQUEST_TEMPLATE.md create mode 100644 .github/markdownlint.yml create mode 100644 .gitignore create mode 100644 .travis.yml create mode 100644 CHANGELOG.md create mode 100644 CODE_OF_CONDUCT.md create mode 100644 Dockerfile create mode 100644 LICENSE create mode 100644 README.md create mode 100644 assets/email_template.html create mode 100644 assets/email_template.txt create mode 100644 assets/multiqc_config.yaml create mode 100644 assets/sendmail_template.txt create mode 100755 bin/markdown_to_html.r create mode 100755 bin/scrape_software_versions.py create mode 100644 conf/awsbatch.config create mode 100644 conf/base.config create mode 100644 conf/igenomes.config create mode 100644 conf/test.config create mode 100644 docs/README.md create mode 100644 docs/output.md create mode 100644 docs/usage.md create mode 100644 environment.yml create mode 100644 main.nf create mode 100644 nextflow.config diff --git a/.gitattributes b/.gitattributes new file mode 100644 index 00000000..7fe55006 --- /dev/null +++ b/.gitattributes @@ -0,0 +1 @@ +*.config linguist-language=nextflow diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md new file mode 100644 index 00000000..6376a3c9 --- /dev/null +++ b/.github/CONTRIBUTING.md @@ -0,0 +1,47 @@ +# nf-core/atacseq: Contributing Guidelines + +Hi there! Many thanks for taking an interest in improving nf-core/atacseq. + +We try to manage the required tasks for nf-core/atacseq using GitHub issues, you probably came to this page when creating one. Please use the pre-filled template to save time. + +However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) + +> If you need help using or modifying nf-core/atacseq then the best place to ask is on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). + + + +## Contribution workflow +If you'd like to write some code for nf-core/atacseq, the standard workflow +is as follows: + +1. Check that there isn't already an issue about your idea in the + [nf-core/atacseq issues](https://github.com/nf-core/atacseq/issues) to avoid + duplicating work. + * If there isn't one already, please create one so that others know you're working on this +2. Fork the [nf-core/atacseq repository](https://github.com/nf-core/atacseq) to your GitHub account +3. Make the necessary changes / additions within your forked repository +4. Submit a Pull Request against the `dev` branch and wait for the code to be reviewed and merged. + +If you're not used to this workflow with git, you can start with some [basic docs from GitHub](https://help.github.com/articles/fork-a-repo/) or even their [excellent interactive tutorial](https://try.github.io/). + + +## Tests +When you create a pull request with changes, [Travis CI](https://travis-ci.org/) will run automatic tests. +Typically, pull-requests are only fully reviewed when these tests are passing, though of course we can help out before then. + +There are typically two types of tests that run: + +### Lint Tests +The nf-core has a [set of guidelines](http://nf-co.re/guidelines) which all pipelines must adhere to. +To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint ` command. + +If any failures or warnings are encountered, please follow the listed URL for more documentation. + +### Pipeline Tests +Each nf-core pipeline should be set up with a minimal set of test-data. +Travis CI then runs the pipeline on this data to ensure that it exists successfully. +If there are any failures then the automated tests fail. +These tests are run both with the latest available version of Nextflow and also the minimum required version that is stated in the pipeline code. + +## Getting help +For further information/help, please consult the [nf-core/atacseq documentation](https://github.com/nf-core/atacseq#documentation) and don't hesitate to get in touch on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). diff --git a/.github/ISSUE_TEMPLATE/bug_report.md b/.github/ISSUE_TEMPLATE/bug_report.md new file mode 100644 index 00000000..b287bce7 --- /dev/null +++ b/.github/ISSUE_TEMPLATE/bug_report.md @@ -0,0 +1,31 @@ +Hi there! + +Thanks for telling us about a problem with the pipeline. Please delete this text and anything that's not relevant from the template below: + +#### Describe the bug +A clear and concise description of what the bug is. + +#### Steps to reproduce +Steps to reproduce the behaviour: +1. Command line: `nextflow run ...` +2. See error: _Please provide your error message_ + +#### Expected behaviour +A clear and concise description of what you expected to happen. + +#### System: + - Hardware: [e.g. HPC, Desktop, Cloud...] + - Executor: [e.g. slurm, local, awsbatch...] + - OS: [e.g. CentOS Linux, macOS, Linux Mint...] + - Version [e.g. 7, 10.13.6, 18.3...] + +#### Nextflow Installation: + - Version: [e.g. 0.31.0] + +#### Container engine: + - Engine: [e.g. Conda, Docker or Singularity] + - version: [e.g. 1.0.0] + - Image tag: [e.g. nfcore/atacseq:1.0.0] + +#### Additional context +Add any other context about the problem here. diff --git a/.github/ISSUE_TEMPLATE/feature_request.md b/.github/ISSUE_TEMPLATE/feature_request.md new file mode 100644 index 00000000..1f025b77 --- /dev/null +++ b/.github/ISSUE_TEMPLATE/feature_request.md @@ -0,0 +1,16 @@ +Hi there! + +Thanks for suggesting a new feature for the pipeline! Please delete this text and anything that's not relevant from the template below: + +#### Is your feature request related to a problem? Please describe. +A clear and concise description of what the problem is. +Ex. I'm always frustrated when [...] + +#### Describe the solution you'd like +A clear and concise description of what you want to happen. + +#### Describe alternatives you've considered +A clear and concise description of any alternative solutions or features you've considered. + +#### Additional context +Add any other context about the feature request here. diff --git a/.github/PULL_REQUEST_TEMPLATE.md b/.github/PULL_REQUEST_TEMPLATE.md new file mode 100644 index 00000000..2784b969 --- /dev/null +++ b/.github/PULL_REQUEST_TEMPLATE.md @@ -0,0 +1,15 @@ +Many thanks to contributing to nf-core/atacseq! + +Please fill in the appropriate checklist below (delete whatever is not relevant). These are the most common things requested on pull requests (PRs). + +## PR checklist + - [ ] This comment contains a description of changes (with reason) + - [ ] If you've fixed a bug or added code that should be tested, add tests! + - [ ] If necessary, also make a PR on the [nf-core/atacseq branch on the nf-core/test-datasets repo]( https://github.com/nf-core/test-datasets/pull/new/nf-core/atacseq) + - [ ] Ensure the test suite passes (`nextflow run . -profile test,docker`). + - [ ] Make sure your code lints (`nf-core lint .`). + - [ ] Documentation in `docs` is updated + - [ ] `CHANGELOG.md` is updated + - [ ] `README.md` is updated + +**Learn more about contributing:** https://github.com/nf-core/atacseq/tree/master/.github/CONTRIBUTING.md diff --git a/.github/markdownlint.yml b/.github/markdownlint.yml new file mode 100644 index 00000000..e052a635 --- /dev/null +++ b/.github/markdownlint.yml @@ -0,0 +1,9 @@ +# Markdownlint configuration file +default: true, +line-length: false +no-multiple-blanks: 0 +blanks-around-headers: false +blanks-around-lists: false +header-increment: false +no-duplicate-header: + siblings_only: true diff --git a/.gitignore b/.gitignore new file mode 100644 index 00000000..5b54e3e6 --- /dev/null +++ b/.gitignore @@ -0,0 +1,7 @@ +.nextflow* +work/ +data/ +results/ +.DS_Store +tests/test_data +*.pyc diff --git a/.travis.yml b/.travis.yml new file mode 100644 index 00000000..b00e5355 --- /dev/null +++ b/.travis.yml @@ -0,0 +1,42 @@ +sudo: required +language: python +jdk: openjdk8 +services: docker +python: '3.6' +cache: pip +matrix: + fast_finish: true + +before_install: + # PRs to master are only ok if coming from dev branch + - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ])' + # Pull the docker image first so the test doesn't wait for this + - docker pull nfcore/atacseq:dev + # Fake the tag locally so that the pipeline runs properly + # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) + - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + +install: + # Install Nextflow + - mkdir /tmp/nextflow && cd /tmp/nextflow + - wget -qO- get.nextflow.io | bash + - sudo ln -s /tmp/nextflow/nextflow /usr/local/bin/nextflow + # Install nf-core/tools + - pip install --upgrade pip + - pip install nf-core + # Reset + - mkdir ${TRAVIS_BUILD_DIR}/tests && cd ${TRAVIS_BUILD_DIR}/tests + # Install markdownlint-cli + - sudo apt-get install npm && npm install -g markdownlint-cli + +env: + - NXF_VER='0.32.0' # Specify a minimum NF version that should be tested and work + - NXF_VER='' # Plus: get the latest NF version and check that it works + +script: + # Lint the pipeline code + - nf-core lint ${TRAVIS_BUILD_DIR} + # Lint the documentation + - markdownlint ${TRAVIS_BUILD_DIR} -c ${TRAVIS_BUILD_DIR}/.github/markdownlint.yml + # Run the pipeline with the test profile + - nextflow run ${TRAVIS_BUILD_DIR} -profile test,docker diff --git a/CHANGELOG.md b/CHANGELOG.md new file mode 100644 index 00000000..bf57ded8 --- /dev/null +++ b/CHANGELOG.md @@ -0,0 +1,4 @@ +# nf-core/atacseq: Changelog + +## v1.0.0 - [date] +Initial release of nf-core/atacseq, created with the [nf-core](http://nf-co.re/) template. diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md new file mode 100644 index 00000000..09226d0d --- /dev/null +++ b/CODE_OF_CONDUCT.md @@ -0,0 +1,46 @@ +# Contributor Covenant Code of Conduct + +## Our Pledge + +In the interest of fostering an open and welcoming environment, we as contributors and maintainers pledge to making participation in our project and our community a harassment-free experience for everyone, regardless of age, body size, disability, ethnicity, gender identity and expression, level of experience, nationality, personal appearance, race, religion, or sexual identity and orientation. + +## Our Standards + +Examples of behavior that contributes to creating a positive environment include: + +* Using welcoming and inclusive language +* Being respectful of differing viewpoints and experiences +* Gracefully accepting constructive criticism +* Focusing on what is best for the community +* Showing empathy towards other community members + +Examples of unacceptable behavior by participants include: + +* The use of sexualized language or imagery and unwelcome sexual attention or advances +* Trolling, insulting/derogatory comments, and personal or political attacks +* Public or private harassment +* Publishing others' private information, such as a physical or electronic address, without explicit permission +* Other conduct which could reasonably be considered inappropriate in a professional setting + +## Our Responsibilities + +Project maintainers are responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behavior. + +Project maintainers have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this Code of Conduct, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful. + +## Scope + +This Code of Conduct applies both within project spaces and in public spaces when an individual is representing the project or its community. Examples of representing a project or community include using an official project e-mail address, posting via an official social media account, or acting as an appointed representative at an online or offline event. Representation of a project may be further defined and clarified by project maintainers. + +## Enforcement + +Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-core-invite.herokuapp.com/). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. + +Project maintainers who do not follow or enforce the Code of Conduct in good faith may face temporary or permanent repercussions as determined by other members of the project's leadership. + +## Attribution + +This Code of Conduct is adapted from the [Contributor Covenant][homepage], version 1.4, available at [http://contributor-covenant.org/version/1/4][version] + +[homepage]: http://contributor-covenant.org +[version]: http://contributor-covenant.org/version/1/4/ diff --git a/Dockerfile b/Dockerfile new file mode 100644 index 00000000..194b4b52 --- /dev/null +++ b/Dockerfile @@ -0,0 +1,7 @@ +FROM nfcore/base +LABEL authors="Harshil Patel" \ + description="Docker image containing all requirements for nf-core/atacseq pipeline" + +COPY environment.yml / +RUN conda env create -f /environment.yml && conda clean -a +ENV PATH /opt/conda/envs/nf-core-atacseq-1.0.0/bin:$PATH diff --git a/LICENSE b/LICENSE new file mode 100644 index 00000000..515e402d --- /dev/null +++ b/LICENSE @@ -0,0 +1,21 @@ +MIT License + +Copyright (c) Harshil Patel + +Permission is hereby granted, free of charge, to any person obtaining a copy +of this software and associated documentation files (the "Software"), to deal +in the Software without restriction, including without limitation the rights +to use, copy, modify, merge, publish, distribute, sublicense, and/or sell +copies of the Software, and to permit persons to whom the Software is +furnished to do so, subject to the following conditions: + +The above copyright notice and this permission notice shall be included in all +copies or substantial portions of the Software. + +THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR +IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, +FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE +AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER +LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, +OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE +SOFTWARE. diff --git a/README.md b/README.md new file mode 100644 index 00000000..28ce2325 --- /dev/null +++ b/README.md @@ -0,0 +1,30 @@ +# nf-core/atacseq + +**ATACSeq peak-calling and differential analysis pipeline.**. + +[![Build Status](https://travis-ci.com/nf-core/atacseq.svg?branch=master)](https://travis-ci.com/nf-core/atacseq) +[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A50.32.0-brightgreen.svg)](https://www.nextflow.io/) + +[![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) +[![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) + +## Introduction +The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It comes with docker containers making installation trivial and results highly reproducible. + + +## Documentation +The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: + +1. [Installation](https://nf-co.re/usage/installation) +2. Pipeline configuration + * [Local installation](https://nf-co.re/usage/local_installation) + * [Adding your own system config](https://nf-co.re/usage/adding_own_config) + * [Reference genomes](https://nf-co.re/usage/reference_genomes) +3. [Running the pipeline](docs/usage.md) +4. [Output and how to interpret the results](docs/output.md) +5. [Troubleshooting](https://nf-co.re/usage/troubleshooting) + + + +## Credits +nf-core/atacseq was originally written by Harshil Patel. diff --git a/assets/email_template.html b/assets/email_template.html new file mode 100644 index 00000000..1c4f6946 --- /dev/null +++ b/assets/email_template.html @@ -0,0 +1,52 @@ + + + + + + + + + nf-core/atacseq Pipeline Report + + +
+ +

nf-core/atacseq v${version}

+

Run Name: $runName

+ +<% if (!success){ + out << """ +
+

nf-core/atacseq execution completed unsuccessfully!

+

The exit status of the task that caused the workflow execution to fail was: $exitStatus.

+

The full error message was:

+ 
${errorReport}
+
+ """ +} else { + out << """ +
+ nf-core/atacseq execution completed successfully! +
+ """ +} +%> + +

The workflow was completed at $dateComplete (duration: $duration)

+

The command used to launch the workflow was as follows:

+
$commandLine
+ +

Pipeline Configuration:

+ + + <% out << summary.collect{ k,v -> "" }.join("\n") %> + +
$k
$v
+ +

nf-core/atacseq

+

https://github.com/nf-core/atacseq

+ +
+ + + diff --git a/assets/email_template.txt b/assets/email_template.txt new file mode 100644 index 00000000..38059dd2 --- /dev/null +++ b/assets/email_template.txt @@ -0,0 +1,34 @@ +======================================== + nf-core/atacseq v${version} +======================================== +Run Name: $runName + +<% if (success){ + out << "## nf-core/atacseq execution completed successfully! ##" +} else { + out << """#################################################### +## nf-core/atacseq execution completed unsuccessfully! ## +#################################################### +The exit status of the task that caused the workflow execution to fail was: $exitStatus. +The full error message was: + +${errorReport} +""" +} %> + + +The workflow was completed at $dateComplete (duration: $duration) + +The command used to launch the workflow was as follows: + + $commandLine + + + +Pipeline Configuration: +----------------------- +<% out << summary.collect{ k,v -> " - $k: $v" }.join("\n") %> + +-- +nf-core/atacseq +https://github.com/nf-core/atacseq diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml new file mode 100644 index 00000000..8e6246d7 --- /dev/null +++ b/assets/multiqc_config.yaml @@ -0,0 +1,9 @@ +report_comment: > + This report has been generated by the nf-core/atacseq + analysis pipeline. For information about how to interpret these results, please see the + documentation. +report_section_order: + nf-core/atacseq-software-versions: + order: -1000 + +export_plots: true diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt new file mode 100644 index 00000000..2d671220 --- /dev/null +++ b/assets/sendmail_template.txt @@ -0,0 +1,36 @@ +To: $email +Subject: $subject +Mime-Version: 1.0 +Content-Type: multipart/related;boundary="nfcoremimeboundary" + +--nfcoremimeboundary +Content-Type: text/html; charset=utf-8 + +$email_html + +<% +if (mqcFile){ +def mqcFileObj = new File("$mqcFile") +if (mqcFileObj.length() < mqcMaxSize){ +out << """ +--nfcoremimeboundary +Content-Type: text/html; name=\"multiqc_report\" +Content-Transfer-Encoding: base64 +Content-ID: +Content-Disposition: attachment; filename=\"${mqcFileObj.getName()}\" + +${mqcFileObj. + bytes. + encodeBase64(). + toString(). + tokenize( '\n' )*. + toList()*. + collate( 76 )*. + collect { it.join() }. + flatten(). + join( '\n' )} +""" +}} +%> + +--nfcoremimeboundary-- diff --git a/bin/markdown_to_html.r b/bin/markdown_to_html.r new file mode 100755 index 00000000..abe13350 --- /dev/null +++ b/bin/markdown_to_html.r @@ -0,0 +1,51 @@ +#!/usr/bin/env Rscript + +# Command line argument processing +args = commandArgs(trailingOnly=TRUE) +if (length(args) < 2) { + stop("Usage: markdown_to_html.r ", call.=FALSE) +} +markdown_fn <- args[1] +output_fn <- args[2] + +# Load / install packages +if (!require("markdown")) { + install.packages("markdown", dependencies=TRUE, repos='http://cloud.r-project.org/') + library("markdown") +} + +base_css_fn <- getOption("markdown.HTML.stylesheet") +base_css <- readChar(base_css_fn, file.info(base_css_fn)$size) +custom_css <- paste(base_css, " +body { + padding: 3em; + margin-right: 350px; + max-width: 100%; +} +#toc { + position: fixed; + right: 20px; + width: 300px; + padding-top: 20px; + overflow: scroll; + height: calc(100% - 3em - 20px); +} +#toc_header { + font-size: 1.8em; + font-weight: bold; +} +#toc > ul { + padding-left: 0; + list-style-type: none; +} +#toc > ul ul { padding-left: 20px; } +#toc > ul > li > a { display: none; } +img { max-width: 800px; } +") + +markdownToHTML( + file = markdown_fn, + output = output_fn, + stylesheet = custom_css, + options = c('toc', 'base64_images', 'highlight_code') +) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py new file mode 100755 index 00000000..d66a7915 --- /dev/null +++ b/bin/scrape_software_versions.py @@ -0,0 +1,49 @@ +#!/usr/bin/env python +from __future__ import print_function +from collections import OrderedDict +import re + +# TODO nf-core: Add additional regexes for new tools in process get_software_versions +regexes = { + 'nf-core/atacseq': ['v_pipeline.txt', r"(\S+)"], + 'Nextflow': ['v_nextflow.txt', r"(\S+)"], + 'FastQC': ['v_fastqc.txt', r"FastQC v(\S+)"], + 'MultiQC': ['v_multiqc.txt', r"multiqc, version (\S+)"], +} +results = OrderedDict() +results['nf-core/atacseq'] = 'N/A' +results['Nextflow'] = 'N/A' +results['FastQC'] = 'N/A' +results['MultiQC'] = 'N/A' + +# Search each file using its regex +for k, v in regexes.items(): + with open(v[0]) as x: + versions = x.read() + match = re.search(v[1], versions) + if match: + results[k] = "v{}".format(match.group(1)) + +# Remove software set to false in results +for k in results: + if not results[k]: + del(results[k]) + +# Dump to YAML +print (''' +id: 'software_versions' +section_name: 'nf-core/atacseq Software Versions' +section_href: 'https://github.com/nf-core/atacseq' +plot_type: 'html' +description: 'are collected at run time from the software output.' +data: | +
+''') +for k,v in results.items(): + print("
{}
{}
".format(k,v)) +print ("
") + +# Write out regexes as csv file: +with open('software_versions.csv', 'w') as f: + for k,v in results.items(): + f.write("{}\t{}\n".format(k,v)) diff --git a/conf/awsbatch.config b/conf/awsbatch.config new file mode 100644 index 00000000..14af5866 --- /dev/null +++ b/conf/awsbatch.config @@ -0,0 +1,18 @@ +/* + * ------------------------------------------------- + * Nextflow config file for running on AWS batch + * ------------------------------------------------- + * Base config needed for running with -profile awsbatch + */ +params { + config_profile_name = 'AWSBATCH' + config_profile_description = 'AWSBATCH Cloud Profile' + config_profile_contact = 'Alexander Peltzer (@apeltzer)' + config_profile_url = 'https://aws.amazon.com/de/batch/' +} + +aws.region = params.awsregion +process.executor = 'awsbatch' +process.queue = params.awsqueue +executor.awscli = '/home/ec2-user/miniconda/bin/aws' +params.tracedir = './' diff --git a/conf/base.config b/conf/base.config new file mode 100644 index 00000000..db12e92d --- /dev/null +++ b/conf/base.config @@ -0,0 +1,34 @@ +/* + * ------------------------------------------------- + * nf-core/atacseq Nextflow base config file + * ------------------------------------------------- + * A 'blank slate' config file, appropriate for general + * use on most high performace compute environments. + * Assumes that all software is installed and available + * on the PATH. Runs in `local` mode - all jobs will be + * run on the logged in environment. + */ + +process { + + // TODO nf-core: Check the defaults for all processes + cpus = { check_max( 1 * task.attempt, 'cpus' ) } + memory = { check_max( 8.GB * task.attempt, 'memory' ) } + time = { check_max( 2.h * task.attempt, 'time' ) } + + errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } + maxRetries = 1 + maxErrors = '-1' + + // Process-specific resource requirements + // TODO nf-core: Customise requirements for specific processes. + // See https://www.nextflow.io/docs/latest/config.html#config-process-selectors +} + +params { + // Defaults only, expecting to be overwritten + max_memory = 128.GB + max_cpus = 16 + max_time = 240.h + igenomes_base = 's3://ngi-igenomes/igenomes/' +} diff --git a/conf/igenomes.config b/conf/igenomes.config new file mode 100644 index 00000000..d19e61f4 --- /dev/null +++ b/conf/igenomes.config @@ -0,0 +1,147 @@ +/* + * ------------------------------------------------- + * Nextflow config file for iGenomes paths + * ------------------------------------------------- + * Defines reference genomes, using iGenome paths + * Can be used by any config that customises the base + * path using $params.igenomes_base / --igenomes_base + */ + +params { + // illumina iGenomes reference file paths + // TODO nf-core: Add new reference types and strip out those that are not needed + genomes { + 'GRCh37' { + bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" + } + 'GRCm38' { + bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" + } + 'TAIR10' { + bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" + } + 'EB2' { + bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" + } + 'UMD3.1' { + bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" + } + 'WBcel235' { + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" + } + 'CanFam3.1' { + bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" + } + 'GRCz10' { + bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" + } + 'BDGP6' { + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" + } + 'EquCab2' { + bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" + } + 'EB1' { + bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" + } + 'Galgal4' { + bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" + } + 'Gm01' { + bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" + } + 'Mmul_1' { + bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" + } + 'IRGSP-1.0' { + bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" + } + 'CHIMP2.1.4' { + bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" + } + 'Rnor_6.0' { + bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" + } + 'R64-1-1' { + bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" + } + 'EF2' { + bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" + } + 'Sbi1' { + bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" + } + 'Sscrofa10.2' { + bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" + } + 'AGPv3' { + bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" + fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" + gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" + star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" + } + } +} diff --git a/conf/test.config b/conf/test.config new file mode 100644 index 00000000..c77002df --- /dev/null +++ b/conf/test.config @@ -0,0 +1,25 @@ +/* + * ------------------------------------------------- + * Nextflow config file for running tests + * ------------------------------------------------- + * Defines bundled input files and everything required + * to run a fast and simple test. Use as follows: + * nextflow run nf-core/atacseq -profile test + */ + +params { + config_profile_name = 'Test profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + // Limit resources so that this can run on Travis + max_cpus = 2 + max_memory = 6.GB + max_time = 48.h + // Input data + // TODO nf-core: Specify the paths to your test data on nf-core/test-datasets + // TODO nf-core: Give any required params for the test so that command line flags are not needed + singleEnd = false + readPaths = [ + ['Testdata', ['https://github.com/nf-core/test-datasets/raw/exoseq/testdata/Testdata_R1.tiny.fastq.gz', 'https://github.com/nf-core/test-datasets/raw/exoseq/testdata/Testdata_R2.tiny.fastq.gz']], + ['SRR389222', ['https://github.com/nf-core/test-datasets/raw/methylseq/testdata/SRR389222_sub1.fastq.gz', 'https://github.com/nf-core/test-datasets/raw/methylseq/testdata/SRR389222_sub2.fastq.gz']] + ] +} diff --git a/docs/README.md b/docs/README.md new file mode 100644 index 00000000..6ebe6ecc --- /dev/null +++ b/docs/README.md @@ -0,0 +1,12 @@ +# nf-core/atacseq: Documentation + +The nf-core/atacseq documentation is split into the following files: + +1. [Installation](https://nf-co.re/usage/installation) +2. Pipeline configuration + * [Local installation](https://nf-co.re/usage/local_installation) + * [Adding your own system config](https://nf-co.re/usage/adding_own_config) + * [Reference genomes](https://nf-co.re/usage/reference_genomes) +3. [Running the pipeline](usage.md) +4. [Output and how to interpret the results](output.md) +5. [Troubleshooting](https://nf-co.re/usage/troubleshooting) diff --git a/docs/output.md b/docs/output.md new file mode 100644 index 00000000..ebed1ce6 --- /dev/null +++ b/docs/output.md @@ -0,0 +1,41 @@ +# nf-core/atacseq: Output + +This document describes the output produced by the pipeline. Most of the plots are taken from the MultiQC report, which summarises results at the end of the pipeline. + + + +## Pipeline overview +The pipeline is built using [Nextflow](https://www.nextflow.io/) +and processes data using the following steps: + +* [FastQC](#fastqc) - read quality control +* [MultiQC](#multiqc) - aggregate report, describing results of the whole pipeline + +## FastQC +[FastQC](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) gives general quality metrics about your reads. It provides information about the quality score distribution across your reads, the per base sequence content (%T/A/G/C). You get information about adapter contamination and other overrepresented sequences. + +For further reading and documentation see the [FastQC help](http://www.bioinformatics.babraham.ac.uk/projects/fastqc/Help/). + +> **NB:** The FastQC plots displayed in the MultiQC report shows _untrimmed_ reads. They may contain adapter sequence and potentially regions with low quality. To see how your reads look after trimming, look at the FastQC reports in the `trim_galore` directory. + +**Output directory: `results/fastqc`** + +* `sample_fastqc.html` + * FastQC report, containing quality metrics for your untrimmed raw fastq files +* `zips/sample_fastqc.zip` + * zip file containing the FastQC report, tab-delimited data file and plot images + + +## MultiQC +[MultiQC](http://multiqc.info) is a visualisation tool that generates a single HTML report summarising all samples in your project. Most of the pipeline QC results are visualised in the report and further statistics are available in within the report data directory. + +The pipeline has special steps which allow the software versions used to be reported in the MultiQC output for future traceability. + +**Output directory: `results/multiqc`** + +* `Project_multiqc_report.html` + * MultiQC report - a standalone HTML file that can be viewed in your web browser +* `Project_multiqc_data/` + * Directory containing parsed statistics from the different tools used in the pipeline + +For more information about how to use MultiQC reports, see [http://multiqc.info](http://multiqc.info) diff --git a/docs/usage.md b/docs/usage.md new file mode 100644 index 00000000..7f9b905b --- /dev/null +++ b/docs/usage.md @@ -0,0 +1,282 @@ +# nf-core/atacseq: Usage + +## Table of contents + + + +* [Table of contents](#table-of-contents) +* [Introduction](#introduction) +* [Running the pipeline](#running-the-pipeline) + * [Updating the pipeline](#updating-the-pipeline) + * [Reproducibility](#reproducibility) +* [Main arguments](#main-arguments) + * [`-profile`](#-profile) + * [`--reads`](#--reads) + * [`--singleEnd`](#--singleend) +* [Reference genomes](#reference-genomes) + * [`--genome` (using iGenomes)](#--genome-using-igenomes) + * [`--fasta`](#--fasta) + * [`--igenomesIgnore`](#--igenomesignore) +* [Job resources](#job-resources) + * [Automatic resubmission](#automatic-resubmission) + * [Custom resource requests](#custom-resource-requests) +* [AWS Batch specific parameters](#aws-batch-specific-parameters) + * [`--awsqueue`](#--awsqueue) + * [`--awsregion`](#--awsregion) +* [Other command line parameters](#other-command-line-parameters) + * [`--outdir`](#--outdir) + * [`--email`](#--email) + * [`-name`](#-name) + * [`-resume`](#-resume) + * [`-c`](#-c) + * [`--custom_config_version`](#--custom_config_version) + * [`--custom_config_base`](#--custom_config_base) + * [`--max_memory`](#--max_memory) + * [`--max_time`](#--max_time) + * [`--max_cpus`](#--max_cpus) + * [`--plaintext_email`](#--plaintext_email) + * [`--monochrome_logs`](#--monochrome_logs) + * [`--multiqc_config`](#--multiqc_config) + + + +## Introduction +Nextflow handles job submissions on SLURM or other environments, and supervises running the jobs. Thus the Nextflow process must run until the pipeline is finished. We recommend that you put the process running in the background through `screen` / `tmux` or similar tool. Alternatively you can run nextflow within a cluster job submitted your job scheduler. + +It is recommended to limit the Nextflow Java virtual machines memory. We recommend adding the following line to your environment (typically in `~/.bashrc` or `~./bash_profile`): + +```bash +NXF_OPTS='-Xms1g -Xmx4g' +``` + + + +## Running the pipeline +The typical command for running the pipeline is as follows: + +```bash +nextflow run nf-core/atacseq --reads '*_R{1,2}.fastq.gz' -profile docker +``` + +This will launch the pipeline with the `docker` configuration profile. See below for more information about profiles. + +Note that the pipeline will create the following files in your working directory: + +```bash +work # Directory containing the nextflow working files +results # Finished results (configurable, see below) +.nextflow_log # Log file from Nextflow +# Other nextflow hidden files, eg. history of pipeline runs and old logs. +``` + +### Updating the pipeline +When you run the above command, Nextflow automatically pulls the pipeline code from GitHub and stores it as a cached version. When running the pipeline after this, it will always use the cached version if available - even if the pipeline has been updated since. To make sure that you're running the latest version of the pipeline, make sure that you regularly update the cached version of the pipeline: + +```bash +nextflow pull nf-core/atacseq +``` + +### Reproducibility +It's a good idea to specify a pipeline version when running the pipeline on your data. This ensures that a specific version of the pipeline code and software are used when you run your pipeline. If you keep using the same tag, you'll be running the same version of the pipeline, even if there have been changes to the code since. + +First, go to the [nf-core/atacseq releases page](https://github.com/nf-core/atacseq/releases) and find the latest version number - numeric only (eg. `1.3.1`). Then specify this when running the pipeline with `-r` (one hyphen) - eg. `-r 1.3.1`. + +This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. + + +## Main arguments + +### `-profile` +Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! + +If `-profile` is not specified at all the pipeline will be run locally and expects all software to be installed and available on the `PATH`. + +* `awsbatch` + * A generic configuration profile to be used with AWS Batch. +* `conda` + * A generic configuration profile to be used with [conda](https://conda.io/docs/) + * Pulls most software from [Bioconda](https://bioconda.github.io/) +* `docker` + * A generic configuration profile to be used with [Docker](http://docker.com/) + * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) +* `singularity` + * A generic configuration profile to be used with [Singularity](http://singularity.lbl.gov/) + * Pulls software from DockerHub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) +* `test` + * A profile with a complete configuration for automated testing + * Includes links to test data so needs no other parameters + + + +### `--reads` +Use this to specify the location of your input FastQ files. For example: + +```bash +--reads 'path/to/data/sample_*_{1,2}.fastq' +``` + +Please note the following requirements: + +1. The path must be enclosed in quotes +2. The path must have at least one `*` wildcard character +3. When using the pipeline with paired end data, the path must use `{1,2}` notation to specify read pairs. + +If left unspecified, a default pattern is used: `data/*{1,2}.fastq.gz` + +### `--singleEnd` +By default, the pipeline expects paired-end data. If you have single-end data, you need to specify `--singleEnd` on the command line when you launch the pipeline. A normal glob pattern, enclosed in quotation marks, can then be used for `--reads`. For example: + +```bash +--singleEnd --reads '*.fastq' +``` + +It is not possible to run a mixture of single-end and paired-end files in one run. + + +## Reference genomes + +The pipeline config files come bundled with paths to the illumina iGenomes reference index files. If running with docker or AWS, the configuration is set up to use the [AWS-iGenomes](https://ewels.github.io/AWS-iGenomes/) resource. + +### `--genome` (using iGenomes) +There are 31 different species supported in the iGenomes references. To run the pipeline, you must specify which to use with the `--genome` flag. + +You can find the keys to specify the genomes in the [iGenomes config file](../conf/igenomes.config). Common genomes that are supported are: + +* Human + * `--genome GRCh37` +* Mouse + * `--genome GRCm38` +* _Drosophila_ + * `--genome BDGP6` +* _S. cerevisiae_ + * `--genome 'R64-1-1'` + +> There are numerous others - check the config file for more. + +Note that you can use the same configuration setup to save sets of reference files for your own use, even if they are not part of the iGenomes resource. See the [Nextflow documentation](https://www.nextflow.io/docs/latest/config.html) for instructions on where to save such a file. + +The syntax for this reference configuration is as follows: + + + +```nextflow +params { + genomes { + 'GRCh37' { + fasta = '' // Used if no star index given + } + // Any number of additional genomes, key is used with --genome + } +} +``` + + +### `--fasta` +If you prefer, you can specify the full path to your reference genome when you run the pipeline: + +```bash +--fasta '[path to Fasta reference]' +``` + +### `--igenomesIgnore` +Do not load `igenomes.config` when running the pipeline. You may choose this option if you observe clashes between custom parameters and those supplied in `igenomes.config`. + +## Job resources +### Automatic resubmission +Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with an error code of `143` (exceeded requested resources) it will automatically resubmit with higher requests (2 x original, then 3 x original). If it still fails after three times then the pipeline is stopped. + +### Custom resource requests +Wherever process-specific requirements are set in the pipeline, the default value can be changed by creating a custom config file. See the files hosted at [`nf-core/configs`](https://github.com/nf-core/configs/tree/master/conf) for examples. + +If you are likely to be running `nf-core` pipelines regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter (see definition below). You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile. + +If you have any questions or issues please send us a message on [Slack](https://nf-core-invite.herokuapp.com/). + +## AWS Batch specific parameters +Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use the `-awsbatch` profile and then specify all of the following parameters. +### `--awsqueue` +The JobQueue that you intend to use on AWS Batch. +### `--awsregion` +The AWS region to run your job in. Default is set to `eu-west-1` but can be adjusted to your needs. + +Please make sure to also set the `-w/--work-dir` and `--outdir` parameters to a S3 storage bucket of your choice - you'll get an error message notifying you if you didn't. + +## Other command line parameters + + + +### `--outdir` +The output directory where the results will be saved. + +### `--email` +Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run. + +### `-name` +Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. + +This is used in the MultiQC report (if not default) and in the summary HTML / e-mail (always). + +**NB:** Single hyphen (core Nextflow option) + +### `-resume` +Specify this when restarting a pipeline. Nextflow will used cached results from any pipeline steps where the inputs are the same, continuing from where it got to previously. + +You can also supply a run name to resume a specific run: `-resume [run-name]`. Use the `nextflow log` command to show previous run names. + +**NB:** Single hyphen (core Nextflow option) + +### `-c` +Specify the path to a specific config file (this is a core NextFlow command). + +**NB:** Single hyphen (core Nextflow option) + +Note - you can use this to override pipeline defaults. + +### `--custom_config_version` +Provide git commit id for custom Institutional configs hosted at `nf-core/configs`. This was implemented for reproducibility purposes. Default is set to `master`. + +```bash +## Download and use config file with following git commid id +--custom_config_version d52db660777c4bf36546ddb188ec530c3ada1b96 +``` + +### `--custom_config_base` +If you're running offline, nextflow will not be able to fetch the institutional config files +from the internet. If you don't need them, then this is not a problem. If you do need them, +you should download the files from the repo and tell nextflow where to find them with the +`custom_config_base` option. For example: + +```bash +## Download and unzip the config files +cd /path/to/my/configs +wget https://github.com/nf-core/configs/archive/master.zip +unzip master.zip + +## Run the pipeline +cd /path/to/my/data +nextflow run /path/to/pipeline/ --custom_config_base /path/to/my/configs/configs-master/ +``` + +> Note that the nf-core/tools helper package has a `download` command to download all required pipeline +> files + singularity containers + institutional configs in one go for you, to make this process easier. + +### `--max_memory` +Use to set a top-limit for the default memory requirement for each process. +Should be a string in the format integer-unit. eg. `--max_memory '8.GB'` + +### `--max_time` +Use to set a top-limit for the default time requirement for each process. +Should be a string in the format integer-unit. eg. `--max_time '2.h'` + +### `--max_cpus` +Use to set a top-limit for the default CPU requirement for each process. +Should be a string in the format integer-unit. eg. `--max_cpus 1` + +### `--plaintext_email` +Set to receive plain-text e-mails instead of HTML formatted. + +### `--monochrome_logs` +Set to disable colourful command line output and live life in monochrome. + +### `--multiqc_config` +Specify a path to a custom MultiQC configuration file. diff --git a/environment.yml b/environment.yml new file mode 100644 index 00000000..2fe4a741 --- /dev/null +++ b/environment.yml @@ -0,0 +1,11 @@ +# You can use this file to create a conda environment for this pipeline: +# conda env create -f environment.yml +name: nf-core-atacseq-1.0.0 +channels: + - conda-forge + - bioconda + - defaults +dependencies: + # TODO nf-core: Add required software dependencies here + - fastqc=0.11.8 + - multiqc=1.7 diff --git a/main.nf b/main.nf new file mode 100644 index 00000000..ff53c27e --- /dev/null +++ b/main.nf @@ -0,0 +1,428 @@ +#!/usr/bin/env nextflow +/* +======================================================================================== + nf-core/atacseq +======================================================================================== + nf-core/atacseq Analysis Pipeline. + #### Homepage / Documentation + https://github.com/nf-core/atacseq +---------------------------------------------------------------------------------------- +*/ + + +def helpMessage() { + // TODO nf-core: Add to this help message with new command line parameters + log.info nfcoreHeader() + log.info""" + + Usage: + + The typical command for running the pipeline is as follows: + + nextflow run nf-core/atacseq --reads '*_R{1,2}.fastq.gz' -profile docker + + Mandatory arguments: + --reads Path to input data (must be surrounded with quotes) + -profile Configuration profile to use. Can use multiple (comma separated) + Available: conda, docker, singularity, awsbatch, test and more. + + Options: + --genome Name of iGenomes reference + --singleEnd Specifies that the input is single end reads + + References If not specified in the configuration file or you wish to overwrite any of the references. + --fasta Path to Fasta reference + + Other options: + --outdir The output directory where the results will be saved + --email Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits + --maxMultiqcEmailFileSize Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) + -name Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. + + AWSBatch options: + --awsqueue The AWSBatch JobQueue that needs to be set when running on AWSBatch + --awsregion The AWS Region for your AWS Batch job to run on + """.stripIndent() +} + +/* + * SET UP CONFIGURATION VARIABLES + */ + +// Show help emssage +if (params.help){ + helpMessage() + exit 0 +} + +// Check if genome exists in the config file +if (params.genomes && params.genome && !params.genomes.containsKey(params.genome)) { + exit 1, "The provided genome '${params.genome}' is not available in the iGenomes file. Currently the available genomes are ${params.genomes.keySet().join(", ")}" +} + +// TODO nf-core: Add any reference files that are needed +// Configurable reference genomes +fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false +if ( params.fasta ){ + fasta = file(params.fasta) + if( !fasta.exists() ) exit 1, "Fasta file not found: ${params.fasta}" +} +// +// NOTE - THIS IS NOT USED IN THIS PIPELINE, EXAMPLE ONLY +// If you want to use the above in a process, define the following: +// input: +// file fasta from fasta +// + + +// Has the run name been specified by the user? +// this has the bonus effect of catching both -name and --name +custom_runName = params.name +if( !(workflow.runName ==~ /[a-z]+_[a-z]+/) ){ + custom_runName = workflow.runName +} + + +if( workflow.profile == 'awsbatch') { + // AWSBatch sanity checking + if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" + // Check outdir paths to be S3 buckets if running on AWSBatch + // related: https://github.com/nextflow-io/nextflow/issues/813 + if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" + // Prevent trace files to be stored on S3 since S3 does not support rolling files. + if (workflow.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." +} + +// Stage config files +ch_multiqc_config = Channel.fromPath(params.multiqc_config) +ch_output_docs = Channel.fromPath("$baseDir/docs/output.md") + +/* + * Create a channel for input read files + */ +if(params.readPaths){ + if(params.singleEnd){ + Channel + .from(params.readPaths) + .map { row -> [ row[0], [file(row[1][0])]] } + .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } + .into { read_files_fastqc; read_files_trimming } + } else { + Channel + .from(params.readPaths) + .map { row -> [ row[0], [file(row[1][0]), file(row[1][1])]] } + .ifEmpty { exit 1, "params.readPaths was empty - no input files supplied" } + .into { read_files_fastqc; read_files_trimming } + } +} else { + Channel + .fromFilePairs( params.reads, size: params.singleEnd ? 1 : 2 ) + .ifEmpty { exit 1, "Cannot find any reads matching: ${params.reads}\nNB: Path needs to be enclosed in quotes!\nIf this is single-end data, please specify --singleEnd on the command line." } + .into { read_files_fastqc; read_files_trimming } +} + + +// Header log info +log.info nfcoreHeader() +def summary = [:] +if(workflow.revision) summary['Pipeline Release'] = workflow.revision +summary['Run Name'] = custom_runName ?: workflow.runName +// TODO nf-core: Report custom parameters here +summary['Reads'] = params.reads +summary['Fasta Ref'] = params.fasta +summary['Data Type'] = params.singleEnd ? 'Single-End' : 'Paired-End' +summary['Max Resources'] = "$params.max_memory memory, $params.max_cpus cpus, $params.max_time time per job" +if(workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" +summary['Output dir'] = params.outdir +summary['Launch dir'] = workflow.launchDir +summary['Working dir'] = workflow.workDir +summary['Script dir'] = workflow.projectDir +summary['User'] = workflow.userName +if(workflow.profile == 'awsbatch'){ + summary['AWS Region'] = params.awsregion + summary['AWS Queue'] = params.awsqueue +} +summary['Config Profile'] = workflow.profile +if(params.config_profile_description) summary['Config Description'] = params.config_profile_description +if(params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact +if(params.config_profile_url) summary['Config URL'] = params.config_profile_url +if(params.email) { + summary['E-mail Address'] = params.email + summary['MultiQC maxsize'] = params.maxMultiqcEmailFileSize +} +log.info summary.collect { k,v -> "${k.padRight(18)}: $v" }.join("\n") +log.info "\033[2m----------------------------------------------------\033[0m" + +// Check the hostnames against configured profiles +checkHostname() + +def create_workflow_summary(summary) { + def yaml_file = workDir.resolve('workflow_summary_mqc.yaml') + yaml_file.text = """ + id: 'nf-core-atacseq-summary' + description: " - this information is collected when the pipeline is started." + section_name: 'nf-core/atacseq Workflow Summary' + section_href: 'https://github.com/nf-core/atacseq' + plot_type: 'html' + data: | +
+${summary.collect { k,v -> "
$k
${v ?: 'N/A'}
" }.join("\n")} +
+ """.stripIndent() + + return yaml_file +} + + +/* + * Parse software version numbers + */ +process get_software_versions { + publishDir "${params.outdir}/pipeline_info", mode: 'copy', + saveAs: {filename -> + if (filename.indexOf(".csv") > 0) filename + else null + } + + output: + file 'software_versions_mqc.yaml' into software_versions_yaml + file "software_versions.csv" + + script: + // TODO nf-core: Get all tools to print their version number here + """ + echo $workflow.manifest.version > v_pipeline.txt + echo $workflow.nextflow.version > v_nextflow.txt + fastqc --version > v_fastqc.txt + multiqc --version > v_multiqc.txt + scrape_software_versions.py &> software_versions_mqc.yaml + """ +} + + + +/* + * STEP 1 - FastQC + */ +process fastqc { + tag "$name" + publishDir "${params.outdir}/fastqc", mode: 'copy', + saveAs: {filename -> filename.indexOf(".zip") > 0 ? "zips/$filename" : "$filename"} + + input: + set val(name), file(reads) from read_files_fastqc + + output: + file "*_fastqc.{zip,html}" into fastqc_results + + script: + """ + fastqc -q $reads + """ +} + + + +/* + * STEP 2 - MultiQC + */ +process multiqc { + publishDir "${params.outdir}/MultiQC", mode: 'copy' + + input: + file multiqc_config from ch_multiqc_config + // TODO nf-core: Add in log files from your new processes for MultiQC to find! + file ('fastqc/*') from fastqc_results.collect().ifEmpty([]) + file ('software_versions/*') from software_versions_yaml.collect() + file workflow_summary from create_workflow_summary(summary) + + output: + file "*multiqc_report.html" into multiqc_report + file "*_data" + file "multiqc_plots" + + script: + rtitle = custom_runName ? "--title \"$custom_runName\"" : '' + rfilename = custom_runName ? "--filename " + custom_runName.replaceAll('\\W','_').replaceAll('_+','_') + "_multiqc_report" : '' + // TODO nf-core: Specify which MultiQC modules to use with -m for a faster run time + """ + multiqc -f $rtitle $rfilename --config $multiqc_config . + """ +} + + + +/* + * STEP 3 - Output Description HTML + */ +process output_documentation { + publishDir "${params.outdir}/pipeline_info", mode: 'copy' + + input: + file output_docs from ch_output_docs + + output: + file "results_description.html" + + script: + """ + markdown_to_html.r $output_docs results_description.html + """ +} + + + +/* + * Completion e-mail notification + */ +workflow.onComplete { + + // Set up the e-mail variables + def subject = "[nf-core/atacseq] Successful: $workflow.runName" + if(!workflow.success){ + subject = "[nf-core/atacseq] FAILED: $workflow.runName" + } + def email_fields = [:] + email_fields['version'] = workflow.manifest.version + email_fields['runName'] = custom_runName ?: workflow.runName + email_fields['success'] = workflow.success + email_fields['dateComplete'] = workflow.complete + email_fields['duration'] = workflow.duration + email_fields['exitStatus'] = workflow.exitStatus + email_fields['errorMessage'] = (workflow.errorMessage ?: 'None') + email_fields['errorReport'] = (workflow.errorReport ?: 'None') + email_fields['commandLine'] = workflow.commandLine + email_fields['projectDir'] = workflow.projectDir + email_fields['summary'] = summary + email_fields['summary']['Date Started'] = workflow.start + email_fields['summary']['Date Completed'] = workflow.complete + email_fields['summary']['Pipeline script file path'] = workflow.scriptFile + email_fields['summary']['Pipeline script hash ID'] = workflow.scriptId + if(workflow.repository) email_fields['summary']['Pipeline repository Git URL'] = workflow.repository + if(workflow.commitId) email_fields['summary']['Pipeline repository Git Commit'] = workflow.commitId + if(workflow.revision) email_fields['summary']['Pipeline Git branch/tag'] = workflow.revision + if(workflow.container) email_fields['summary']['Docker image'] = workflow.container + email_fields['summary']['Nextflow Version'] = workflow.nextflow.version + email_fields['summary']['Nextflow Build'] = workflow.nextflow.build + email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp + + // TODO nf-core: If not using MultiQC, strip out this code (including params.maxMultiqcEmailFileSize) + // On success try attach the multiqc report + def mqc_report = null + try { + if (workflow.success) { + mqc_report = multiqc_report.getVal() + if (mqc_report.getClass() == ArrayList){ + log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" + mqc_report = mqc_report[0] + } + } + } catch (all) { + log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" + } + + // Render the TXT template + def engine = new groovy.text.GStringTemplateEngine() + def tf = new File("$baseDir/assets/email_template.txt") + def txt_template = engine.createTemplate(tf).make(email_fields) + def email_txt = txt_template.toString() + + // Render the HTML template + def hf = new File("$baseDir/assets/email_template.html") + def html_template = engine.createTemplate(hf).make(email_fields) + def email_html = html_template.toString() + + // Render the sendmail template + def smail_fields = [ email: params.email, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.maxMultiqcEmailFileSize.toBytes() ] + def sf = new File("$baseDir/assets/sendmail_template.txt") + def sendmail_template = engine.createTemplate(sf).make(smail_fields) + def sendmail_html = sendmail_template.toString() + + // Send the HTML e-mail + if (params.email) { + try { + if( params.plaintext_email ){ throw GroovyException('Send plaintext e-mail, not HTML') } + // Try to send HTML e-mail using sendmail + [ 'sendmail', '-t' ].execute() << sendmail_html + log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (sendmail)" + } catch (all) { + // Catch failures and try with plaintext + [ 'mail', '-s', subject, params.email ].execute() << email_txt + log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (mail)" + } + } + + // Write summary e-mail HTML to a file + def output_d = new File( "${params.outdir}/pipeline_info/" ) + if( !output_d.exists() ) { + output_d.mkdirs() + } + def output_hf = new File( output_d, "pipeline_report.html" ) + output_hf.withWriter { w -> w << email_html } + def output_tf = new File( output_d, "pipeline_report.txt" ) + output_tf.withWriter { w -> w << email_txt } + + c_reset = params.monochrome_logs ? '' : "\033[0m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; + c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_red = params.monochrome_logs ? '' : "\033[0;31m"; + + if (workflow.stats.ignoredCountFmt > 0 && workflow.success) { + log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" + log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCountFmt} ${c_reset}" + log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCountFmt} ${c_reset}" + } + + if(workflow.success){ + log.info "${c_purple}[nf-core/atacseq]${c_green} Pipeline completed successfully${c_reset}" + } else { + checkHostname() + log.info "${c_purple}[nf-core/atacseq]${c_red} Pipeline completed with errors${c_reset}" + } + +} + + +def nfcoreHeader(){ + // Log colors ANSI codes + c_reset = params.monochrome_logs ? '' : "\033[0m"; + c_dim = params.monochrome_logs ? '' : "\033[2m"; + c_black = params.monochrome_logs ? '' : "\033[0;30m"; + c_green = params.monochrome_logs ? '' : "\033[0;32m"; + c_yellow = params.monochrome_logs ? '' : "\033[0;33m"; + c_blue = params.monochrome_logs ? '' : "\033[0;34m"; + c_purple = params.monochrome_logs ? '' : "\033[0;35m"; + c_cyan = params.monochrome_logs ? '' : "\033[0;36m"; + c_white = params.monochrome_logs ? '' : "\033[0;37m"; + + return """ ${c_dim}----------------------------------------------------${c_reset} + ${c_green},--.${c_black}/${c_green},-.${c_reset} + ${c_blue} ___ __ __ __ ___ ${c_green}/,-._.--~\'${c_reset} + ${c_blue} |\\ | |__ __ / ` / \\ |__) |__ ${c_yellow}} {${c_reset} + ${c_blue} | \\| | \\__, \\__/ | \\ |___ ${c_green}\\`-._,-`-,${c_reset} + ${c_green}`._,._,\'${c_reset} + ${c_purple} nf-core/atacseq v${workflow.manifest.version}${c_reset} + ${c_dim}----------------------------------------------------${c_reset} + """.stripIndent() +} + +def checkHostname(){ + def c_reset = params.monochrome_logs ? '' : "\033[0m" + def c_white = params.monochrome_logs ? '' : "\033[0;37m" + def c_red = params.monochrome_logs ? '' : "\033[1;91m" + def c_yellow_bold = params.monochrome_logs ? '' : "\033[1;93m" + if(params.hostnames){ + def hostname = "hostname".execute().text.trim() + params.hostnames.each { prof, hnames -> + hnames.each { hname -> + if(hostname.contains(hname) && !workflow.profile.contains(prof)){ + log.error "====================================================\n" + + " ${c_red}WARNING!${c_reset} You are running with `-profile $workflow.profile`\n" + + " but your machine hostname is ${c_white}'$hostname'${c_reset}\n" + + " ${c_yellow_bold}It's highly recommended that you use `-profile $prof${c_reset}`\n" + + "============================================================" + } + } + } + } +} diff --git a/nextflow.config b/nextflow.config new file mode 100644 index 00000000..050b5d8f --- /dev/null +++ b/nextflow.config @@ -0,0 +1,127 @@ +/* + * ------------------------------------------------- + * nf-core/atacseq Nextflow config file + * ------------------------------------------------- + * Default config options for all environments. + */ + +// Global default params, used in configs +params { + + // Workflow flags + // TODO nf-core: Specify your pipeline's command line flags + reads = "data/*{1,2}.fastq.gz" + singleEnd = false + outdir = './results' + + // Boilerplate options + name = false + multiqc_config = "$baseDir/assets/multiqc_config.yaml" + email = false + maxMultiqcEmailFileSize = 25.MB + plaintext_email = false + monochrome_logs = false + help = false + igenomes_base = "./iGenomes" + tracedir = "${params.outdir}/pipeline_info" + awsqueue = false + awsregion = 'eu-west-1' + igenomesIgnore = false + custom_config_version = 'master' + custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" + hostnames = false + config_profile_description = false + config_profile_contact = false + config_profile_url = false +} + +// Container slug. Stable releases should specify release tag! +// Developmental code should specify :dev +process.container = 'nfcore/atacseq:dev' + +// Load base.config by default for all pipelines +includeConfig 'conf/base.config' + +// Load nf-core custom profiles from different Institutions +try { + includeConfig "${params.custom_config_base}/nfcore_custom.config" +} catch (Exception e) { + System.err.println("WARNING: Could not load nf-core/config profiles: ${params.custom_config_base}/nfcore_custom.config") +} + +profiles { + awsbatch { includeConfig 'conf/awsbatch.config' } + conda { process.conda = "$baseDir/environment.yml" } + debug { process.beforeScript = 'echo $HOSTNAME' } + docker { docker.enabled = true } + singularity { singularity.enabled = true } + test { includeConfig 'conf/test.config' } +} + +// Load igenomes.config if required +if(!params.igenomesIgnore){ + includeConfig 'conf/igenomes.config' +} + +// Capture exit codes from upstream processes when piping +process.shell = ['/bin/bash', '-euo', 'pipefail'] + +timeline { + enabled = true + file = "${params.tracedir}/execution_timeline.html" +} +report { + enabled = true + file = "${params.tracedir}/execution_report.html" +} +trace { + enabled = true + file = "${params.tracedir}/execution_trace.txt" +} +dag { + enabled = true + file = "${params.tracedir}/pipeline_dag.svg" +} + +manifest { + name = 'nf-core/atacseq' + author = 'Harshil Patel' + homePage = 'https://github.com/nf-core/atacseq' + description = 'ATACSeq peak-calling and differential analysis pipeline.' + mainScript = 'main.nf' + nextflowVersion = '>=0.32.0' + version = '1.0.0' +} + +// Function to ensure that resource requirements don't go beyond +// a maximum limit +def check_max(obj, type) { + if(type == 'memory'){ + try { + if(obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) + return params.max_memory as nextflow.util.MemoryUnit + else + return obj + } catch (all) { + println " ### ERROR ### Max memory '${params.max_memory}' is not valid! Using default value: $obj" + return obj + } + } else if(type == 'time'){ + try { + if(obj.compareTo(params.max_time as nextflow.util.Duration) == 1) + return params.max_time as nextflow.util.Duration + else + return obj + } catch (all) { + println " ### ERROR ### Max time '${params.max_time}' is not valid! Using default value: $obj" + return obj + } + } else if(type == 'cpus'){ + try { + return Math.min( obj, params.max_cpus as int ) + } catch (all) { + println " ### ERROR ### Max cpus '${params.max_cpus}' is not valid! Using default value: $obj" + return obj + } + } +} From 337ebfad2dfef18268ee1f7afb75cfc16b9c0f35 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 11 Apr 2019 14:15:03 +0100 Subject: [PATCH 002/149] Update to current version in dev --- nextflow.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/nextflow.config b/nextflow.config index fb96e8a2..dd0764f1 100755 --- a/nextflow.config +++ b/nextflow.config @@ -69,7 +69,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:dev' +process.container = 'nfcore/atacseq:1.0.0' // Load base.config by default for all pipelines includeConfig 'conf/base.config' From ca4fc2f143ebd5f3c20612219074ced0bb9d8702 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 11 Apr 2019 14:22:47 +0100 Subject: [PATCH 003/149] @drpatelh --- .travis.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.travis.yml b/.travis.yml index b00e5355..110d7d41 100755 --- a/.travis.yml +++ b/.travis.yml @@ -14,7 +14,7 @@ before_install: - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) - - docker tag nfcore/atacseq:dev nfcore/atacseq:dev + - docker tag nfcore/atacseq:dev nfcore/atacseq:1.0.0 install: # Install Nextflow From dcb222086673d5f915de0000cfbfb7c11e3fbe84 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 11 Apr 2019 15:50:07 +0100 Subject: [PATCH 004/149] Bump version to 1.0.1dev --- .travis.yml | 2 +- CHANGELOG.md | 18 ++++++++++++++++-- Dockerfile | 2 +- conf/test.config | 3 +++ docs/output.md | 2 +- environment.yml | 2 +- main.nf | 2 +- nextflow.config | 4 ++-- 8 files changed, 26 insertions(+), 9 deletions(-) diff --git a/.travis.yml b/.travis.yml index 110d7d41..b00e5355 100755 --- a/.travis.yml +++ b/.travis.yml @@ -14,7 +14,7 @@ before_install: - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly # Looks weird when this is :dev to :dev, but makes sense when testing code for a release (:dev to :1.0.1) - - docker tag nfcore/atacseq:dev nfcore/atacseq:1.0.0 + - docker tag nfcore/atacseq:dev nfcore/atacseq:dev install: # Install Nextflow diff --git a/CHANGELOG.md b/CHANGELOG.md index bf57ded8..c5744b4f 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -1,4 +1,18 @@ # nf-core/atacseq: Changelog -## v1.0.0 - [date] -Initial release of nf-core/atacseq, created with the [nf-core](http://nf-co.re/) template. +All notable changes to this project will be documented in this file. + +The format is based on [Keep a Changelog](http://keepachangelog.com/en/1.0.0/) +and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0.html). + +## [Unpublished Version / DEV] + +### `Added` + +* Merged in TEMPLATE branch for automated syncing + +## [1.0.0] - 2019-04-09 + +### `Added` +Initial release of nf-core/atacseq +Created with version 1.1 of the [nf-core](http://nf-co.re/) template. diff --git a/Dockerfile b/Dockerfile index 194b4b52..c8a96fc1 100755 --- a/Dockerfile +++ b/Dockerfile @@ -4,4 +4,4 @@ LABEL authors="Harshil Patel" \ COPY environment.yml / RUN conda env create -f /environment.yml && conda clean -a -ENV PATH /opt/conda/envs/nf-core-atacseq-1.0.0/bin:$PATH +ENV PATH /opt/conda/envs/nf-core-atacseq-1.0.1dev/bin:$PATH diff --git a/conf/test.config b/conf/test.config index d5e12189..9ed9db45 100755 --- a/conf/test.config +++ b/conf/test.config @@ -8,6 +8,9 @@ */ params { + config_profile_name = 'Test profile' + config_profile_description = 'Minimal test dataset to check pipeline function' + max_cpus = 2 max_memory = 6.GB max_time = 12.h diff --git a/docs/output.md b/docs/output.md index a25625f5..ae530a0e 100755 --- a/docs/output.md +++ b/docs/output.md @@ -54,7 +54,7 @@ The initial QC and alignments are performed at the library-level e.g. if the sam [BWA](https://sourceforge.net/projects/bio-bwa/files/), [SAMtools](https://sourceforge.net/projects/samtools/files/samtools/) *Description*: - Adapter-trimmed reads are mapped to the reference assembly using BWA. A genome index is required to run BWA so if this isnt provided explicitly using the `--bwa_index_dir` and `--bwa_index_base` parameters then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--saveGenomeIndex` parameter to save the indices for future pipeline runs, reducing processing times. + Adapter-trimmed reads are mapped to the reference assembly using BWA. A genome index is required to run BWA so if this isnt provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--saveGenomeIndex` parameter to save the indices for future pipeline runs, reducing processing times. File names in the resulting directory (i.e. `bwa/library/`) will have the '`.Lb.`' (**L**i**b**rary) suffix. diff --git a/environment.yml b/environment.yml index 0e032c0e..e18a23a4 100755 --- a/environment.yml +++ b/environment.yml @@ -1,6 +1,6 @@ # You can use this file to create a conda environment for this pipeline: # conda env create -f environment.yml -name: nf-core-atacseq-1.0.0 +name: nf-core-atacseq-1.0.1dev channels: - conda-forge - bioconda diff --git a/main.nf b/main.nf index 454af553..7fc64584 100755 --- a/main.nf +++ b/main.nf @@ -839,7 +839,7 @@ if (params.singleEnd){ /////////////////////////////////////////////////////////////////////////////// /* - * STEP 4.4.1 Picard CollectMultipleMetrics at after merging libraries + * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries */ process merge_library_collectmetrics { tag "$name" diff --git a/nextflow.config b/nextflow.config index dd0764f1..cc0f62c4 100755 --- a/nextflow.config +++ b/nextflow.config @@ -69,7 +69,7 @@ params { // Container slug. Stable releases should specify release tag! // Developmental code should specify :dev -process.container = 'nfcore/atacseq:1.0.0' +process.container = 'nfcore/atacseq:dev' // Load base.config by default for all pipelines includeConfig 'conf/base.config' @@ -127,7 +127,7 @@ manifest { description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' nextflowVersion = '>=0.32.0' - version = '1.0.0' + version = '1.0.1dev' } // Function to ensure that resource requirements don't go beyond From 578250e0e40ffba2f9f8c04d9cea6a9119828ce6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sat, 13 Apr 2019 13:18:00 +0100 Subject: [PATCH 005/149] Renamed channels and processes --- CHANGELOG.md | 2 + bin/{reformat_design.py => check_design.py} | 2 + main.nf | 647 +++++++++----------- 3 files changed, 305 insertions(+), 346 deletions(-) rename bin/{reformat_design.py => check_design.py} (99%) diff --git a/CHANGELOG.md b/CHANGELOG.md index c5744b4f..dbd2b272 100755 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,6 +10,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * Merged in TEMPLATE branch for automated syncing +* Use `ch_` prefix for all channels +* Process renaming to differentiate between nf-core `tools` and pipeline ## [1.0.0] - 2019-04-09 diff --git a/bin/reformat_design.py b/bin/check_design.py similarity index 99% rename from bin/reformat_design.py rename to bin/check_design.py index cf7aafa2..5ef3c657 100755 --- a/bin/reformat_design.py +++ b/bin/check_design.py @@ -141,6 +141,8 @@ def reformat_design(DesignFileIn,DesignFileOut): if len(set(numRepList)) == 1 and multiGroups and repsExist: balancedDesign = True + print repsExist, multiGroups, balancedDesign + ############################################ ############################################ ## RUN FUNCTION diff --git a/main.nf b/main.nf index 7fc64584..2f30851f 100755 --- a/main.nf +++ b/main.nf @@ -124,103 +124,57 @@ if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)){ //////////////////////////////////////////////////// // Pipeline config -output_docs_ch = Channel.fromPath("$baseDir/docs/output.md", checkIfExists: true) +ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) // JSON files required by BAMTools for alignment filtering if (params.singleEnd) { - bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_se_config, checkIfExists: true) + ch_bamtools_filter_config = file(params.bamtools_filter_se_config, checkIfExists: true) } else { - bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_pe_config, checkIfExists: true) + ch_bamtools_filter_config = file(params.bamtools_filter_pe_config, checkIfExists: true) } // Header files for MultiQC -multiqc_config_ch = Channel.fromPath(params.multiqc_config, checkIfExists: true) -mlib_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) -mlib_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) -mlib_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) -mlib_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) -mlib_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) - -mrep_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) -mrep_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) -mrep_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) -mrep_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) -mrep_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) +ch_multiqc_config = file(params.multiqc_config, checkIfExists: true) +ch_mlib_peak_count_header = file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) +ch_mlib_frip_score_header = file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) +ch_mlib_peak_annotation_header = file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) +ch_mlib_deseq2_pca_header = file("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) +ch_mlib_deseq2_clustering_header = file("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) + +ch_mrep_peak_count_header = file("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) +ch_mrep_frip_score_header = file("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) +ch_mrep_peak_annotation_header = file("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) +ch_mrep_deseq2_pca_header = file("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) +ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) //////////////////////////////////////////////////// /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// // Validate inputs -if (params.design){ - design_csv = Channel - .fromPath(params.design, checkIfExists: true) - .ifEmpty { exit 1, "Samples design file not found: ${params.design}" } -} else { - exit 1, "Samples design file not specified!" -} +if (params.design) { ch_design = file(params.design, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } +if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } +if (params.gene_bed) { ch_gene_bed = file(params.gene.bed, checkIfExists: true) } +if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } +if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } if (params.fasta){ lastPath = params.fasta.lastIndexOf(File.separator) bwa_base = params.fasta.substring(lastPath+1) - - Channel - .fromPath(params.fasta, checkIfExists: true) - .ifEmpty { exit 1, "Fasta file not found: ${params.fasta}" } - .into { fasta_bwa_index; - fasta_genome_filter; - fasta_markdup_metrics; - fasta_mlib_metrics; - fasta_mlib_macs_annotate; - fasta_mlib_macs_consensus_annotate; - fasta_mrep_macs_annotate; - fasta_mrep_macs_consensus_annotate; - fasta_igv } + ch_fasta = file(params.fasta, checkIfExists: true) } else { exit 1, "Fasta file not specified!" } -if (params.gtf){ - Channel - .fromPath(params.gtf, checkIfExists: true) - .ifEmpty { exit 1, "GTF annotation file not found: ${params.gtf}" } - .into { gtf_gene_bed; - gtf_mlib_macs_annotate; - gtf_mlib_macs_consensus_annotate; - gtf_mrep_macs_annotate; - gtf_mrep_macs_consensus_annotate } -} else { - exit 1, "GTF annotation file not specified!" -} - if (params.bwa_index){ lastPath = params.bwa_index.lastIndexOf(File.separator) bwa_dir = params.bwa_index.substring(0,lastPath+1) bwa_base = params.bwa_index.substring(lastPath+1) - - bwa_index = Channel + ch_bwa_index = Channel .fromPath(bwa_dir, checkIfExists: true) .ifEmpty { exit 1, "BWA index directory not found: ${bwa_dir}" } } -if (params.gene_bed){ - gene_bed = Channel - .fromPath(params.gene_bed, checkIfExists: true) - .ifEmpty { exit 1, "Gene BED annotation file not found: ${params.gene_bed}" } -} - -if (params.tss_bed){ - tss_bed = Channel - .fromPath(params.tss_bed, checkIfExists: true) - .ifEmpty { exit 1, "TSS BED annotation file not found: ${params.tss_bed}" } -} - -if (params.blacklist) { - blacklist = Channel - .fromPath(params.blacklist, checkIfExists: true) - .ifEmpty { exit 1, "Blacklist file not found: ${params.blacklist}" } -} - //////////////////////////////////////////////////// /* -- AWS -- */ //////////////////////////////////////////////////// @@ -321,18 +275,19 @@ if (!params.macs_gsize){ /* * PREPROCESSING - REFORMAT DESIGN FILE AND CHECK VALIDITY */ -process reformat_design { +process check_design { tag "$design" input: - file design from design_csv + file design from ch_design output: - file "*.csv" into reformat_design + stdout into ch_design_bool + file "*.csv" into ch_design_csv script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ - reformat_design.py $design design_reformat.csv + check_design.py $design ${design.baseName}.reformat.csv """ } @@ -340,34 +295,17 @@ process reformat_design { * Create channels for input fastq files */ if (params.singleEnd) { - reformat_design.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + ch_design_csv.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } + .into { ch_raw_reads_fastqc; + ch_raw_reads_trimgalore } } else { - reformat_design.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + ch_design_csv.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } + .into { ch_raw_reads_fastqc; + ch_raw_reads_trimgalore } } -// Boolean value for replicates existing in design -replicates_exist = design_replicates_exist.map { it -> it[0].split('_')[-2].replaceAll('R','').toInteger() } - .flatten() - .max() - .val > 1 - -// Boolean value for multiple samples existing in design -multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].join('_') } - .flatten() - .unique() - .count() - .val > 1 - /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// /* -- -- */ @@ -387,10 +325,10 @@ if (!params.bwa_index){ saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' input: - file fasta from fasta_bwa_index + file fasta from ch_fasta output: - file "BWAIndex" into bwa_index + file "BWAIndex" into ch_bwa_index script: """ @@ -409,10 +347,10 @@ if (!params.gene_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file gtf from gtf_gene_bed + file gtf from ch_gtf output: - file "*.bed" into gene_bed + file "*.bed" into ch_gene_bed script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -430,10 +368,10 @@ if (!params.tss_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file bed from gene_bed + file bed from ch_gene_bed output: - file "*.bed" into tss_bed + file "*.bed" into ch_tss_bed script: """ @@ -450,15 +388,15 @@ process makeGenomeFilter { publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file fasta from fasta_genome_filter + file fasta from ch_fasta output: - file "$fasta" into genome_fasta // FASTA FILE FOR IGV - file "*.fai" into genome_fai // FAI INDEX FOR REFERENCE GENOME - file "*.txt" into genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV - file "*.bed" into genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING - file "*.sizes" into genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS - genome_sizes_mrep_bigwig + file "$fasta" into ch_genome_fasta // FASTA FILE FOR IGV + file "*.fai" into ch_genome_fai // FAI INDEX FOR REFERENCE GENOME + file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV + file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING + file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS + ch_genome_sizes_mrep_bigwig script: blacklist_filter = params.blacklist ? "sortBed -i ${params.blacklist} -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" @@ -490,10 +428,10 @@ process fastqc { saveAs: {filename -> filename.endsWith(".zip") ? "zips/$filename" : "$filename"} input: - set val(name), file(reads) from raw_reads_fastqc + set val(name), file(reads) from ch_raw_reads_fastqc output: - file "*.{zip,html}" into fastqc_reports_mqc + file "*.{zip,html}" into ch_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -524,11 +462,11 @@ process fastqc { * STEP 2 - Trim Galore! */ if (params.skipTrimming){ - trimmed_reads = raw_reads_trimgalore - trimgalore_results_mqc = [] - trimgalore_fastqc_reports_mqc = [] + ch_trimmed_reads = ch_raw_reads_trimgalore + ch_trimgalore_results_mqc = [] + ch_trimgalore_fastqc_reports_mqc = [] } else { - process trim_galore { + process trimGalore { tag "$name" label 'process_long' publishDir "${params.outdir}/trim_galore", mode: 'copy', @@ -540,12 +478,12 @@ if (params.skipTrimming){ } input: - set val(name), file(reads) from raw_reads_trimgalore + set val(name), file(reads) from ch_raw_reads_trimgalore output: - set val(name), file("*.fq.gz") into trimmed_reads - file "*.txt" into trimgalore_results_mqc - file "*.{zip,html}" into trimgalore_fastqc_reports_mqc + set val(name), file("*.fq.gz") into ch_trimmed_reads + file "*.txt" into ch_trimgalore_results_mqc + file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -579,16 +517,16 @@ if (params.skipTrimming){ /* * STEP 3.1 - Align read 1 with bwa */ -process bwa_mem { +process bwaMEM { tag "$name" label 'process_big' input: - set val(name), file(reads) from trimmed_reads - file index from bwa_index.first() + set val(name), file(reads) from ch_trimmed_reads + file index from ch_bwa_index output: - set val(name), file("*.bam") into bwa_bam + set val(name), file("*.bam") into ch_bwa_bam script: prefix="${name}.Lb" @@ -601,7 +539,7 @@ process bwa_mem { /* * STEP 3.2 - Convert .bam to coordinate sorted .bam */ -process sort_bam { +process sortBAM { tag "$name" label 'process_medium' if (params.saveAlignedIntermediates) { @@ -614,11 +552,11 @@ process sort_bam { } input: - set val(name), file(bam) from bwa_bam + set val(name), file(bam) from ch_bwa_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into sort_bam_mlib - file "*.{flagstat,idxstats,stats}" into sort_bam_flagstat_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_mlib + file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc script: prefix="${name}.Lb" @@ -642,12 +580,12 @@ process sort_bam { /* * STEP 4.1 Merge BAM files for all libraries from same replicate */ -sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { sort_bam_mlib } +ch_sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_sort_bam_mlib } -process merge_library { +process mergeLibBAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -660,13 +598,13 @@ process merge_library { } input: - set val(name), file(bams) from sort_bam_mlib + set val(name), file(bams) from ch_sort_bam_mlib output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mlib_bam_filter, - mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into mlib_bam_stats_mqc - file "*.txt" into mlib_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, + ch_mlib_bam_ataqv + file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc + file "*.txt" into ch_mlib_metrics_mqc script: prefix="${name}.mLb.mkD" @@ -723,7 +661,7 @@ process merge_library { /* * STEP 4.2 Filter BAM file at merged library-level */ -process merge_library_filter { +process mergedLibfilterBAM { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -738,14 +676,14 @@ process merge_library_filter { } input: - set val(name), file(bam) from mlib_bam_filter - file bed from genome_filter_regions.collect() - file bamtools_filter_config from bamtools_filter_config_ch.collect() + set val(name), file(bam) from ch_mlib_bam_filter + file bed from ch_genome_filter_regions.collect() + file bamtools_filter_config from ch_bamtools_filter_config output: - set val(name), file("*.{bam,bam.bai}") into mlib_filter_bam - set val(name), file("*.flagstat") into mlib_filter_flagstat - file "*.{idxstats,stats}" into mlib_filter_stats_mqc + set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam + set val(name), file("*.flagstat") into ch_mlib_filter_flagstat + file "*.{idxstats,stats}" into ch_mlib_filter_stats_mqc script: prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" @@ -777,18 +715,18 @@ process merge_library_filter { * STEP 4.3 Remove orphan reads from paired-end BAM file */ if (params.singleEnd){ - mlib_filter_bam.into { mlib_rm_orphan_bam_metrics; - mlib_rm_orphan_bam_bigwig; - mlib_rm_orphan_bam_macs; - mlib_rm_orphan_bam_mrep; - mlib_name_bam_mlib_counts; - mlib_name_bam_mrep_counts } - mlib_filter_flagstat.into { mlib_rm_orphan_flagstat_bigwig; - mlib_rm_orphan_flagstat_macs; - mlib_rm_orphan_flagstat_mqc } - mlib_filter_stats_mqc.set { mlib_rm_orphan_stats_mqc } + ch_mlib_filter_bam.into { ch_mlib_rm_orphan_bam_metrics; + ch_mlib_rm_orphan_bam_bigwig; + ch_mlib_rm_orphan_bam_macs; + ch_mlib_rm_orphan_bam_mrep; + ch_mlib_name_bam_mlib_counts; + ch_mlib_name_bam_mrep_counts } + ch_mlib_filter_flagstat.into { ch_mlib_rm_orphan_flagstat_bigwig; + ch_mlib_rm_orphan_flagstat_macs; + ch_mlib_rm_orphan_flagstat_mqc } + ch_mlib_filter_stats_mqc.set { ch_mlib_rm_orphan_stats_mqc } } else { - process merge_library_rm_orphan { + process mergedLibRMOrphanReads { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -802,19 +740,19 @@ if (params.singleEnd){ } input: - set val(name), file(bam) from mlib_filter_bam + set val(name), file(bam) from ch_mlib_filter_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into mlib_rm_orphan_bam_metrics, - mlib_rm_orphan_bam_bigwig, - mlib_rm_orphan_bam_macs, - mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into mlib_name_bam_mlib_counts, - mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into mlib_rm_orphan_flagstat_bigwig, - mlib_rm_orphan_flagstat_macs, - mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into mlib_rm_orphan_stats_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, + ch_mlib_rm_orphan_bam_bigwig, + ch_mlib_rm_orphan_bam_macs, + ch_mlib_rm_orphan_bam_mrep + set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, + ch_mlib_name_bam_mrep_counts + set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, + ch_mlib_rm_orphan_flagstat_macs, + ch_mlib_rm_orphan_flagstat_mqc + file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ prefix="${name}.mLb.clN" @@ -841,7 +779,7 @@ if (params.singleEnd){ /* * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries */ -process merge_library_collectmetrics { +process mergedLibCollectMultipleMetrics { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -852,12 +790,12 @@ process merge_library_collectmetrics { } input: - set val(name), file(bam) from mlib_rm_orphan_bam_metrics - file fasta from fasta_mlib_metrics.collect() + set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics + file fasta from ch_fasta output: - file "*_metrics" into mlib_collectmetrics_mqc - file "*.pdf" into mlib_collectmetrics_pdf + file "*_metrics" into ch_mlib_collectmetrics_mqc + file "*.pdf" into ch_mlib_collectmetrics_pdf script: prefix="${name}.mLb.clN" @@ -880,7 +818,7 @@ process merge_library_collectmetrics { /* * STEP 4.4.2 Read depth normalised bigWig */ -process merge_library_bigwig { +process mergedLibBigWig { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', @@ -891,12 +829,12 @@ process merge_library_bigwig { } input: - set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_bigwig.join(mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from genome_sizes_mlib_bigwig.collect() + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mlib_bigwig.collect() output: - file "*.bigWig" into mlib_bigwig_igv - file "*.txt" into mlib_bigwig_scale + file "*.bigWig" into ch_mlib_bigwig_igv + file "*.txt" into ch_mlib_bigwig_scale script: prefix="${name}.mLb.clN" @@ -914,7 +852,7 @@ process merge_library_bigwig { /* * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score */ -process merge_library_macs { +process mergedLibMACSCallPeak { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy', @@ -924,18 +862,18 @@ process merge_library_macs { } input: - set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_macs.join(mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from mlib_peak_count_header_ch.collect() - file mlib_frip_score_header from mlib_frip_score_header_ch.collect() + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) + file mlib_peak_count_header from ch_mlib_peak_count_header + file mlib_frip_score_header from ch_mlib_frip_score_header output: - set val(name), file("*.{bed,xls,gappedPeak}") into mlib_macs_output - set val(name), file("*$peakext") into mlib_macs_peaks_homer, - mlib_macs_peaks_qc, - mlib_macs_peaks_consensus, - mlib_macs_peaks_ataqv, - mlib_macs_peaks_igv - file "*_mqc.tsv" into mlib_macs_peaks_mqc + set val(name), file("*.{bed,xls,gappedPeak}") into ch_mlib_macs_output + set val(name), file("*$peakext") into ch_mlib_macs_peaks_homer, + ch_mlib_macs_peaks_qc, + ch_mlib_macs_peaks_consensus, + ch_mlib_macs_peaks_ataqv, + ch_mlib_macs_peaks_igv + file "*_mqc.tsv" into ch_mlib_macs_peaks_mqc when: params.macs_gsize @@ -964,17 +902,17 @@ process merge_library_macs { /* * STEP 4.5.2 Annotate peaks with HOMER */ -process merge_library_macs_annotate { +process mergedLibAnnotatePeaks { tag "$name" publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy' input: - set val(name), file(peak) from mlib_macs_peaks_homer - file fasta from fasta_mlib_macs_annotate.collect() - file gtf from gtf_mlib_macs_annotate.collect() + set val(name), file(peak) from ch_mlib_macs_peaks_homer + file fasta from ch_fasta + file gtf from ch_gtf output: - file "*.txt" into mlib_macs_annotate + file "*.txt" into ch_mlib_macs_annotate when: params.macs_gsize @@ -992,18 +930,18 @@ process merge_library_macs_annotate { /* * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process merge_library_macs_qc { +process mergedLibPeakQC { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/qc", mode: 'copy' input: - file peaks from mlib_macs_peaks_qc.collect{ it[1] } - file annos from mlib_macs_annotate.collect() - file mlib_peak_annotation_header from mlib_peak_annotation_header_ch.collect() + file peaks from ch_mlib_macs_peaks_qc.collect{ it[1] } + file annos from ch_mlib_macs_annotate.collect() + file mlib_peak_annotation_header from ch_mlib_peak_annotation_header output: - file "*.{txt,pdf}" into mlib_macs_qc - file "*.tsv" into mlib_macs_qc_mqc + file "*.{txt,pdf}" into ch_mlib_macs_qc + file "*.tsv" into ch_mlib_macs_qc_mqc when: params.macs_gsize @@ -1025,23 +963,30 @@ process merge_library_macs_qc { """ } +// Get boolean values for replicatesExist, multipleGroups, and balancedDesign from checkDesign process +def design_flags(ch) { + ch.map { it.trim().toLowerCase().tokenize() } + .map { it -> [ it[0].asBoolean(), it[1].asBoolean(), it[2].asBoolean() ] } +} + /* * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process merge_library_macs_consensus { +process mergedLibConsensusPeaks { publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: - file peaks from mlib_macs_peaks_consensus.collect{ it[1] } + file peaks from ch_mlib_macs_peaks_consensus.collect{ it[1] } + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.bed" into mlib_macs_consensus_bed, - mlib_macs_consensus_bed_igv - file "*.boolean.txt" into mlib_macs_consensus_bool - file "*.saf" into mlib_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into mlib_macs_consensus_intersect + file "*.bed" into ch_mlib_macs_consensus_bed, + ch_mlib_macs_consensus_bed_igv + file "*.boolean.txt" into ch_mlib_macs_consensus_bool + file "*.saf" into ch_mlib_macs_consensus_saf + file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect - when: params.macs_gsize && (multiple_samples || replicates_exist) + when: params.macs_gsize && (replicatesExist || multipleGroups) script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mLb.clN' @@ -1073,19 +1018,20 @@ process merge_library_macs_consensus { /* * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process merge_library_macs_consensus_annotate { +process mergedLibConsensusPeaksAnnotate { publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: - file bed from mlib_macs_consensus_bed - file bool from mlib_macs_consensus_bool - file fasta from fasta_mlib_macs_consensus_annotate - file gtf from gtf_mlib_macs_consensus_annotate + file bed from ch_mlib_macs_consensus_bed + file bool from ch_mlib_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: file "*.annotatePeaks.txt" into mlib_macs_consensus_annotate - when: params.macs_gsize && (multiple_samples || replicates_exist) + when: params.macs_gsize && (replicatesExist || multipleGroups) script: prefix="consensus_peaks.mLb.clN" @@ -1104,26 +1050,27 @@ process merge_library_macs_consensus_annotate { /* * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process merge_library_macs_consensus_deseq { +process mergedLibConsensusPeaksDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus/deseq2", mode: 'copy' input: - file bams from mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from mlib_macs_consensus_saf.collect() - file mlib_deseq2_pca_header from mlib_deseq2_pca_header_ch.collect() - file mlib_deseq2_clustering_header from mlib_deseq2_clustering_header_ch.collect() + file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } + file saf from ch_mlib_macs_consensus_saf.collect() + file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*featureCounts.txt" into mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into mlib_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into mlib_macs_consensus_deseq_results - file "sizeFactors" into mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into mlib_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into mlib_macs_consensus_deseq_mqc + file "*featureCounts.txt" into ch_mlib_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results + file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed_igv + file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc - when: params.macs_gsize && multiple_samples && replicates_exist + when: params.macs_gsize && replicatesExist && multipleGroups script: prefix="consensus_peaks.mLb.clN" @@ -1150,18 +1097,18 @@ process merge_library_macs_consensus_deseq { /* * STEP 4.6.1 Run ataqv on BAM file and corresponding peaks */ -process merge_library_ataqv { +process mergedLibAtaqv { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' input: - set val(name), file(bam), file(peak) from mlib_bam_ataqv.join(mlib_macs_peaks_ataqv, by: [0]) - file tss_bed from tss_bed.collect() - file autosomes from genome_autosomes.collect() + set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_peaks_ataqv, by: [0]) + file tss_bed from ch_tss_bed.collect() + file autosomes from ch_genome_autosomes.collect() output: - file "*.json" into mlib_ataqv + file "*.json" into ch_mlib_ataqv script: peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" @@ -1183,15 +1130,15 @@ process merge_library_ataqv { /* * STEP 4.6.2 run ataqv mkarv on all JSON files to render web app */ -process merge_library_ataqv_mkarv { +process mergedLibMkarv { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' input: - file json from mlib_ataqv.collect() + file json from ch_mlib_ataqv.collect() output: - file "html" into mlib_ataqv_mkarv + file "html" into ch_mlib_ataqv_mkarv script: """ @@ -1213,12 +1160,12 @@ process merge_library_ataqv_mkarv { /* * STEP 5.1 Merge library BAM files across all replicates */ -mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { mlib_rm_orphan_bam_mrep } +ch_mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_mlib_rm_orphan_bam_mrep } -process merge_replicate { +process mergeRepBAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', @@ -1231,18 +1178,19 @@ process merge_replicate { } input: - set val(name), file(bams) from mlib_rm_orphan_bam_mrep + set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mrep_bam_bigwig, - mrep_bam_macs - set val(name), file("*.flagstat") into mrep_flagstat_bigwig, - mrep_flagstat_macs, - mrep_flagstat_mqc - file "*.{idxstats,stats}" into mrep_stats_mqc - file "*.txt" into mrep_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, + ch_mrep_bam_macs + set val(name), file("*.flagstat") into ch_mrep_flagstat_bigwig, + ch_mrep_flagstat_macs, + ch_mrep_flagstat_mqc + file "*.{idxstats,stats}" into ch_mrep_stats_mqc + file "*.txt" into ch_mrep_metrics_mqc - when: !params.skipMergeReplicates && replicates_exist + when: !params.skipMergeReplicates && replicatesExist script: prefix="${name}.mRp.clN" @@ -1300,7 +1248,7 @@ process merge_replicate { /* * STEP 5.2.1 Read depth normalised bigWig */ -process merge_replicate_bigwig { +process mergedRepBigWig { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', @@ -1311,14 +1259,15 @@ process merge_replicate_bigwig { } input: - set val(name), file(bam), file(flagstat) from mrep_bam_bigwig.join(mrep_flagstat_bigwig, by: [0]) - file sizes from genome_sizes_mrep_bigwig.collect() + set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mrep_bigwig.collect() + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.bigWig" into mrep_bigwig_igv - file "*.txt" into mrep_bigwig_scale + file "*.bigWig" into ch_mrep_bigwig_igv + file "*.txt" into ch_mrep_bigwig_scale - when: !params.skipMergeReplicates && replicates_exist + when: !params.skipMergeReplicates && replicatesExist script: prefix="${name}.mRp.clN" @@ -1336,7 +1285,7 @@ process merge_replicate_bigwig { /* * STEP 5.2.2 Call peaks with MACS2 and calculate FRiP score */ -process merge_replicate_macs { +process mergedRepMACSCallPeak { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy', @@ -1346,19 +1295,20 @@ process merge_replicate_macs { } input: - set val(name), file(bam), file(flagstat) from mrep_bam_macs.join(mrep_flagstat_macs, by: [0]) - file mrep_peak_count_header from mrep_peak_count_header_ch.collect() - file mrep_frip_score_header from mrep_frip_score_header_ch.collect() + set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_flagstat_macs, by: [0]) + file mrep_peak_count_header from ch_mrep_peak_count_header + file mrep_frip_score_header from ch_mrep_frip_score_header + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.{bed,xls,gappedPeak}" into mrep_macs_output - set val(name), file("*$peakext") into mrep_macs_peaks_homer, - mrep_macs_peaks_qc, - mrep_macs_peaks_consensus, - mrep_macs_peaks_igv - file "*_mqc.tsv" into mrep_macs_peak_mqc + file "*.{bed,xls,gappedPeak}" into ch_mrep_macs_output + set val(name), file("*$peakext") into ch_mrep_macs_peaks_homer, + ch_mrep_macs_peaks_qc, + ch_mrep_macs_peaks_consensus, + ch_mrep_macs_peaks_igv + file "*_mqc.tsv" into ch_mrep_macs_peak_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist script: prefix="${name}.mRp.clN" @@ -1385,19 +1335,20 @@ process merge_replicate_macs { /* * STEP 5.2.3 Annotate peaks with HOMER */ -process merge_replicate_macs_annotate { +process mergedRepAnnotatePeaks { tag "$name" publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy' input: - set val(name), file(peak) from mrep_macs_peaks_homer - file fasta from fasta_mrep_macs_annotate.collect() - file gtf from gtf_mrep_macs_annotate.collect() + set val(name), file(peak) from ch_mrep_macs_peaks_homer + file fasta from ch_fasta + file gtf from ch_gtf + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.txt" into mrep_macs_annotate + file "*.txt" into ch_mrep_macs_annotate - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist script: prefix="${name}.mRp.clN" @@ -1413,20 +1364,21 @@ process merge_replicate_macs_annotate { /* * STEP 5.2.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process merge_replicate_macs_qc { +process mergedRepPeakQC { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/qc", mode: 'copy' input: - file peaks from mrep_macs_peaks_qc.collect{ it[1] } - file annos from mrep_macs_annotate.collect() - file mrep_peak_annotation_header from mrep_peak_annotation_header_ch.collect() + file peaks from ch_mrep_macs_peaks_qc.collect{ it[1] } + file annos from ch_mrep_macs_annotate.collect() + file mrep_peak_annotation_header from ch_mrep_peak_annotation_header + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.{txt,pdf}" into mrep_macs_qc - file "*.tsv" into mrep_macs_qc_mqc + file "*.{txt,pdf}" into ch_mrep_macs_qc + file "*.tsv" into ch_mrep_macs_qc_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mRp.clN' @@ -1449,20 +1401,21 @@ process merge_replicate_macs_qc { /* * STEP 5.2.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process merge_replicate_macs_consensus { +process mergedRepConsensusPeaks { publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: - file peaks from mrep_macs_peaks_consensus.collect{ it[1] } + file peaks from ch_mrep_macs_peaks_consensus.collect{ it[1] } + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.bed" into mrep_macs_consensus_bed, - mrep_macs_consensus_bed_igv - file "*.boolean.txt" into mrep_macs_consensus_bool - file "*.saf" into mrep_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into mrep_macs_consensus_intersect + file "*.bed" into ch_mrep_macs_consensus_bed, + ch_mrep_macs_consensus_bed_igv + file "*.boolean.txt" into ch_mrep_macs_consensus_bool + file "*.saf" into ch_mrep_macs_consensus_saf + file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mRp.clN' @@ -1494,19 +1447,20 @@ process merge_replicate_macs_consensus { /* * STEP 5.2.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process merge_replicate_macs_consensus_annotate { +process mergedRepConsensusPeaksAnnotate { publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: - file bed from mrep_macs_consensus_bed - file bool from mrep_macs_consensus_bool - file fasta from fasta_mrep_macs_consensus_annotate - file gtf from gtf_mrep_macs_consensus_annotate + file bed from ch_mrep_macs_consensus_bed + file bool from ch_mrep_macs_consensus_bool + file fasta from ch_fasta + file gtf from ch_gtf + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*.annotatePeaks.txt" into mrep_macs_consensus_annotate + file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups script: prefix="consensus_peaks.mRp.clN" @@ -1525,26 +1479,27 @@ process merge_replicate_macs_consensus_annotate { /* * STEP 5.2.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process merge_replicate_macs_consensus_deseq { +process mergedRepConsensusPeaksDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus/deseq2", mode: 'copy' input: - file bams from mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from mrep_macs_consensus_saf.collect() - file mrep_deseq2_pca_header from mrep_deseq2_pca_header_ch.collect() - file mrep_deseq2_clustering_header from mrep_deseq2_clustering_header_ch.collect() + file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } + file saf from ch_mrep_macs_consensus_saf.collect() + file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header + set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) output: - file "*featureCounts.txt" into mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into mrep_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into mrep_macs_consensus_deseq_results - file "sizeFactors" into mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into mrep_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into mrep_macs_consensus_deseq_mqc + file "*featureCounts.txt" into ch_mrep_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results + file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed_igv + file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups script: prefix="consensus_peaks.mRp.clN" @@ -1583,20 +1538,20 @@ process igv { publishDir "${params.outdir}/igv", mode: 'copy' input: - file fasta from fasta_igv.collect() + file fasta from ch_fasta - file ('bwa/mergedLibrary/bigwig/*') from mlib_bigwig_igv.collect() - file ('bwa/mergedLibrary/macs/*') from mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/*') from mlib_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/deseq2/*') from mlib_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedLibrary/bigwig/*') from ch_mlib_bigwig_igv.collect() + file ('bwa/mergedLibrary/macs/*') from ch_mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) + file ('bwa/mergedLibrary/macs/consensus/*') from ch_mlib_macs_consensus_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedLibrary/macs/consensus/deseq2/*') from ch_mlib_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/bigwig/*') from mrep_bigwig_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/*') from mrep_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/*') from mrep_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/deseq2/*') from mrep_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/bigwig/*') from ch_mrep_bigwig_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/macs/*') from ch_mrep_macs_peaks_igv.collect{it[1]}.ifEmpty([]) + file ('bwa/mergedReplicate/macs/consensus/*') from ch_mrep_macs_consensus_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/macs/consensus/deseq2/*') from ch_mrep_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) output: - file "*.{txt,xml}" into igv_session + file "*.{txt,xml}" into ch_igv_session script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ outdir_abspath = new File(params.outdir).getCanonicalPath().toString() @@ -1626,7 +1581,7 @@ process get_software_versions { } output: - file 'software_versions_mqc.yaml' into software_versions_mqc + file 'software_versions_mqc.yaml' into ch_software_versions_mqc file "software_versions.csv" script: @@ -1676,37 +1631,37 @@ process multiqc { publishDir "${params.outdir}/multiqc", mode: 'copy' input: - file multiqc_config from multiqc_config_ch.collect() - - file ('fastqc/*') from fastqc_reports_mqc.collect() - file ('trimgalore/*') from trimgalore_results_mqc.collect() - file ('trimgalore/fastqc/*') from trimgalore_fastqc_reports_mqc.collect() - - file ('alignment/library/*') from sort_bam_flagstat_mqc.collect() - - file ('alignment/mergedLibrary/*') from mlib_bam_stats_mqc.collect() - file ('alignment/mergedLibrary/*') from mlib_rm_orphan_flagstat_mqc.collect{it[1]} - file ('alignment/mergedLibrary/*') from mlib_rm_orphan_stats_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from mlib_metrics_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from mlib_collectmetrics_mqc.collect() - file ('macs/mergedLibrary/*') from mlib_macs_peaks_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/*') from mlib_macs_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - file ('alignment/mergedReplicate/*') from mrep_flagstat_mqc.collect{it[1]}.ifEmpty([]) - file ('alignment/mergedReplicate/*') from mrep_stats_mqc.collect().ifEmpty([]) - file ('alignment/mergedReplicate/*') from mrep_metrics_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from mrep_macs_peak_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from mrep_macs_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - file ('software_versions/*') from software_versions_mqc.collect() + file multiqc_config from ch_multiqc_config + + file ('fastqc/*') from ch_fastqc_reports_mqc.collect() + file ('trimgalore/*') from ch_trimgalore_results_mqc.collect() + file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect() + + file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() + + file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} + file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_metrics_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() + file ('macs/mergedLibrary/*') from ch_mlib_macs_peaks_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/*') from ch_mlib_macs_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + file ('alignment/mergedReplicate/*') from ch_mrep_flagstat_mqc.collect{it[1]}.ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_stats_mqc.collect().ifEmpty([]) + file ('alignment/mergedReplicate/*') from ch_mrep_metrics_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_macs_peak_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from ch_mrep_macs_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + file ('software_versions/*') from ch_software_versions_mqc.collect() file ('workflow_summary/*') from create_workflow_summary(summary) output: - file "*multiqc_report.html" into multiqc_report + file "*multiqc_report.html" into ch_multiqc_report file "*_data" file "multiqc_plots" @@ -1734,7 +1689,7 @@ process output_documentation { publishDir "${params.outdir}/Documentation", mode: 'copy' input: - file output_docs from output_docs_ch + file output_docs from ch_output_docs output: file "results_description.html" From 672391a7f91311945c323ff97c9079589cf0a55b Mon Sep 17 00:00:00 2001 From: drpatelh Date: Wed, 26 Jun 2019 13:00:17 +0100 Subject: [PATCH 006/149] Update files --- README.md | 8 ++++++++ assets/nfcore-atacseq_logo.png | Bin 14846 -> 10910 bytes bin/check_design.py | 18 +++--------------- main.nf | 2 +- 4 files changed, 12 insertions(+), 16 deletions(-) diff --git a/README.md b/README.md index 9c853408..1f34cf82 100755 --- a/README.md +++ b/README.md @@ -6,6 +6,7 @@ [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) +[![DOI](https://zenodo.org/badge/156259074.svg)](https://zenodo.org/badge/latestdoi/156259074) ### Introduction @@ -74,3 +75,10 @@ The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). The [nf-core/rnaseq](https://github.com/nf-core/rnaseq) and [nf-core/chipseq](https://github.com/nf-core/chipseq) pipelines developed by Phil Ewels were initially used as a template for this pipeline. Many thanks to Phil for all of his help and advice, and the team at SciLifeLab. Many thanks to others who have helped out along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@sven1103](https://github.com/sven1103), [@MaxUlysse](https://github.com/MaxUlysse), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso). + +## Citation + +If you use nf-core/atacseq for your analysis, please cite it using the following doi: [10.5281/zenodo.2634132](https://doi.org/10.5281/zenodo.2634132) + +You can cite the `nf-core` pre-print as follows: +Ewels PA, Peltzer A, Fillinger S, Alneberg JA, Patel H, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. **nf-core: Community curated bioinformatics pipelines**. *bioRxiv*. 2019. p. 610741. 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zV?720@8u}_n}sm38#*YX#8qFjjN@lWrm|_dOAB(77=TBIL}`p=K_YfE0s&B>pRDM# z7wzb~zB*tWv)C5R0q;{_s;4LbyDO1Lpx2XzAO#_{RGWJ5{$XXv1`$EHRL?~Nb zE>XGnO$t3GDKqs>^Y9vdCG8SzOQ@B5NCY3?j-sb*Ux<4JA0-&qL#y3Qlu&zbBGn$q zc1+;g%8-18VlCS6k%T$Yu|hXWemPIohk?xe5aW@o0S70ay8J--^3QIZ{FY{-gVS*GlW0k6EpnV#go(Eow5QEHd4;@-B8nfUK9^AlQDwGUe^S1R z{66s;D}m8YQv7fM8i)o*hhRf=BrLG*{+n@qvGaDHSe|>1v9mIh7BF5St1IyWJTQt;Z@tURL|w4 z@qp@??mCTZ?_n9i4N-iPIP!3=6knwbH^h6FCZ """ +Epilog = """Example usage: python check_design.py """ argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) @@ -33,7 +33,7 @@ ############################################ ############################################ -def reformat_design(DesignFileIn,DesignFileOut): +def check_design(DesignFileIn,DesignFileOut): ERROR_STR = 'ERROR: Please check design file' HEADER = ['group', 'replicate', 'fastq_1', 'fastq_2'] @@ -76,16 +76,6 @@ def reformat_design(DesignFileIn,DesignFileOut): print "{}: FastQ file has incorrect extension (has to be '.fastq.gz' or 'fq.gz') - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) sys.exit(1) - ## CHECK FASTQ FILES EXIST PER SAMPLE - if fastq[:4] not in ['http']: - if not os.path.exists(fastq): - print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) - sys.exit(1) - else: - if requests.head(fastq).status_code >= 400: - print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) - sys.exit(1) - ## CREATE GROUP MAPPING DICT = {GROUP_ID: {REPLICATE_ID:[[FASTQ_FILES]]} replicate = int(replicate) if not groupRepDict.has_key(group): @@ -141,15 +131,13 @@ def reformat_design(DesignFileIn,DesignFileOut): if len(set(numRepList)) == 1 and multiGroups and repsExist: balancedDesign = True - print repsExist, multiGroups, balancedDesign - ############################################ ############################################ ## RUN FUNCTION ############################################ ############################################ -reformat_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) +check_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) ############################################ ############################################ diff --git a/main.nf b/main.nf index 2f30851f..239ce95f 100755 --- a/main.nf +++ b/main.nf @@ -523,7 +523,7 @@ process bwaMEM { input: set val(name), file(reads) from ch_trimmed_reads - file index from ch_bwa_index + file index from ch_bwa_index.collect() output: set val(name), file("*.bam") into ch_bwa_bam From 13a60de387299fa8564eaed1ab53388f52afe73f Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Sep 2019 14:15:53 +0100 Subject: [PATCH 007/149] Fix tyop --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 1f34cf82..d98a2b3c 100755 --- a/README.md +++ b/README.md @@ -68,7 +68,7 @@ The nf-core/atacseq pipeline comes with documentation about the pipeline, found ### Credits -The pipeline was originally written by the [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. +The pipeline was originally written by [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). From a1dd21c0d5e9f981a5bfd40ffb563335257823d6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 27 Sep 2019 14:21:14 +0100 Subject: [PATCH 008/149] Fix bullets --- README.md | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/README.md b/README.md index d98a2b3c..b1014840 100755 --- a/README.md +++ b/README.md @@ -8,13 +8,13 @@ [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) [![DOI](https://zenodo.org/badge/156259074.svg)](https://zenodo.org/badge/latestdoi/156259074) -### Introduction +## Introduction **nfcore/atacseq** is a bioinformatics analysis pipeline used for ATAC-seq data. The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It comes with docker containers making installation trivial and results highly reproducible. -### Pipeline summary +## Pipeline summary 1. Raw read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) 2. Adapter trimming ([`Trim Galore!`](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/)) @@ -54,7 +54,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool 7. Create IGV session file containing bigWig tracks, peaks and differential sites for data visualisation ([`IGV`](https://software.broadinstitute.org/software/igv/)). 8. Present QC for raw read, alignment, peak-calling and differential accessibility results ([`ataqv`](https://github.com/ParkerLab/ataqv), [`MultiQC`](http://multiqc.info/), [`R`](https://www.r-project.org/)) -### Documentation +## Documentation The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: 1. [Installation](https://nf-co.re/usage/installation) @@ -66,7 +66,7 @@ The nf-core/atacseq pipeline comes with documentation about the pipeline, found 4. [Output and how to interpret the results](docs/output.md) 5. [Troubleshooting](https://nf-co.re/usage/troubleshooting) -### Credits +## Credits The pipeline was originally written by [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. From efaf08a5b0fa941787f08f93719e270e657d5547 Mon Sep 17 00:00:00 2001 From: Mingda Jin Date: Fri, 27 Sep 2019 17:22:43 -0700 Subject: [PATCH 009/149] Use staged blacklist file in makeGenomeFilter --- main.nf | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 5a732493..498501cb 100755 --- a/main.nf +++ b/main.nf @@ -451,6 +451,7 @@ process makeGenomeFilter { input: file fasta from fasta_genome_filter + file blacklist from blacklist output: file "$fasta" into genome_fasta // FASTA FILE FOR IGV @@ -461,7 +462,7 @@ process makeGenomeFilter { genome_sizes_mrep_bigwig script: - blacklist_filter = params.blacklist ? "sortBed -i ${params.blacklist} -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" + blacklist_filter = params.blacklist ? "sortBed -i ${blacklist} -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" mito_filter = params.keepMito ? "" : name_filter """ From 2ffcd46f59803fc7169967f5202c9d225ad72121 Mon Sep 17 00:00:00 2001 From: Mingda Jin Date: Fri, 27 Sep 2019 18:19:25 -0700 Subject: [PATCH 010/149] Make sure channel variable blacklist is declared Create an empty channel for blacklist when params.blacklist is false. --- main.nf | 2 ++ 1 file changed, 2 insertions(+) diff --git a/main.nf b/main.nf index 498501cb..6c68c4c9 100755 --- a/main.nf +++ b/main.nf @@ -219,6 +219,8 @@ if (params.blacklist) { blacklist = Channel .fromPath(params.blacklist, checkIfExists: true) .ifEmpty { exit 1, "Blacklist file not found: ${params.blacklist}" } +} else { + blacklist = Channel.empty() } //////////////////////////////////////////////////// From fc5c79de9ed9b4476a70423ca27a7cad22ff3c54 Mon Sep 17 00:00:00 2001 From: Mingda Jin Date: Sat, 28 Sep 2019 23:10:33 -0700 Subject: [PATCH 011/149] Allocate more RAM to the process running MACS2 --- main.nf | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/main.nf b/main.nf index 5a732493..0e5d5f4e 100755 --- a/main.nf +++ b/main.nf @@ -916,7 +916,7 @@ process merge_library_bigwig { */ process merge_library_macs { tag "$name" - label 'process_long' + label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy', saveAs: {filename -> if (filename.endsWith(".tsv")) "qc/$filename" @@ -1338,7 +1338,7 @@ process merge_replicate_bigwig { */ process merge_replicate_macs { tag "$name" - label 'process_long' + label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy', saveAs: {filename -> if (filename.endsWith(".tsv")) "qc/$filename" From c4a001358da6f6670caeec4f98bb259e513fe43e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:10:07 +0100 Subject: [PATCH 012/149] Revert to dev --- CHANGELOG.md | 2 - README.md | 18 +- bin/{check_design.py => reformat_design.py} | 16 +- main.nf | 647 +++++++++++--------- 4 files changed, 364 insertions(+), 319 deletions(-) mode change 100755 => 100644 CHANGELOG.md mode change 100755 => 100644 README.md rename bin/{check_design.py => reformat_design.py} (87%) mode change 100755 => 100644 mode change 100755 => 100644 main.nf diff --git a/CHANGELOG.md b/CHANGELOG.md old mode 100755 new mode 100644 index dbd2b272..c5744b4f --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,8 +10,6 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * Merged in TEMPLATE branch for automated syncing -* Use `ch_` prefix for all channels -* Process renaming to differentiate between nf-core `tools` and pipeline ## [1.0.0] - 2019-04-09 diff --git a/README.md b/README.md old mode 100755 new mode 100644 index b1014840..9c853408 --- a/README.md +++ b/README.md @@ -6,15 +6,14 @@ [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) -[![DOI](https://zenodo.org/badge/156259074.svg)](https://zenodo.org/badge/latestdoi/156259074) -## Introduction +### Introduction **nfcore/atacseq** is a bioinformatics analysis pipeline used for ATAC-seq data. The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It comes with docker containers making installation trivial and results highly reproducible. -## Pipeline summary +### Pipeline summary 1. Raw read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) 2. Adapter trimming ([`Trim Galore!`](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/)) @@ -54,7 +53,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool 7. Create IGV session file containing bigWig tracks, peaks and differential sites for data visualisation ([`IGV`](https://software.broadinstitute.org/software/igv/)). 8. Present QC for raw read, alignment, peak-calling and differential accessibility results ([`ataqv`](https://github.com/ParkerLab/ataqv), [`MultiQC`](http://multiqc.info/), [`R`](https://www.r-project.org/)) -## Documentation +### Documentation The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: 1. [Installation](https://nf-co.re/usage/installation) @@ -66,19 +65,12 @@ The nf-core/atacseq pipeline comes with documentation about the pipeline, found 4. [Output and how to interpret the results](docs/output.md) 5. [Troubleshooting](https://nf-co.re/usage/troubleshooting) -## Credits +### Credits -The pipeline was originally written by [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. +The pipeline was originally written by the [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). The [nf-core/rnaseq](https://github.com/nf-core/rnaseq) and [nf-core/chipseq](https://github.com/nf-core/chipseq) pipelines developed by Phil Ewels were initially used as a template for this pipeline. Many thanks to Phil for all of his help and advice, and the team at SciLifeLab. Many thanks to others who have helped out along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@sven1103](https://github.com/sven1103), [@MaxUlysse](https://github.com/MaxUlysse), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso). - -## Citation - -If you use nf-core/atacseq for your analysis, please cite it using the following doi: [10.5281/zenodo.2634132](https://doi.org/10.5281/zenodo.2634132) - -You can cite the `nf-core` pre-print as follows: -Ewels PA, Peltzer A, Fillinger S, Alneberg JA, Patel H, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. **nf-core: Community curated bioinformatics pipelines**. *bioRxiv*. 2019. p. 610741. [doi: 10.1101/610741](https://www.biorxiv.org/content/10.1101/610741v1). diff --git a/bin/check_design.py b/bin/reformat_design.py old mode 100755 new mode 100644 similarity index 87% rename from bin/check_design.py rename to bin/reformat_design.py index 0afa4ee5..cf7aafa2 --- a/bin/check_design.py +++ b/bin/reformat_design.py @@ -18,7 +18,7 @@ ############################################ Description = 'Reformat nf-core/atacseq design file and check its contents.' -Epilog = """Example usage: python check_design.py """ +Epilog = """Example usage: python reformat_design.py """ argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) @@ -33,7 +33,7 @@ ############################################ ############################################ -def check_design(DesignFileIn,DesignFileOut): +def reformat_design(DesignFileIn,DesignFileOut): ERROR_STR = 'ERROR: Please check design file' HEADER = ['group', 'replicate', 'fastq_1', 'fastq_2'] @@ -76,6 +76,16 @@ def check_design(DesignFileIn,DesignFileOut): print "{}: FastQ file has incorrect extension (has to be '.fastq.gz' or 'fq.gz') - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) sys.exit(1) + ## CHECK FASTQ FILES EXIST PER SAMPLE + if fastq[:4] not in ['http']: + if not os.path.exists(fastq): + print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) + sys.exit(1) + else: + if requests.head(fastq).status_code >= 400: + print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) + sys.exit(1) + ## CREATE GROUP MAPPING DICT = {GROUP_ID: {REPLICATE_ID:[[FASTQ_FILES]]} replicate = int(replicate) if not groupRepDict.has_key(group): @@ -137,7 +147,7 @@ def check_design(DesignFileIn,DesignFileOut): ############################################ ############################################ -check_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) +reformat_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) ############################################ ############################################ diff --git a/main.nf b/main.nf old mode 100755 new mode 100644 index 239ce95f..7fc64584 --- a/main.nf +++ b/main.nf @@ -124,57 +124,103 @@ if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)){ //////////////////////////////////////////////////// // Pipeline config -ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) +output_docs_ch = Channel.fromPath("$baseDir/docs/output.md", checkIfExists: true) // JSON files required by BAMTools for alignment filtering if (params.singleEnd) { - ch_bamtools_filter_config = file(params.bamtools_filter_se_config, checkIfExists: true) + bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_se_config, checkIfExists: true) } else { - ch_bamtools_filter_config = file(params.bamtools_filter_pe_config, checkIfExists: true) + bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_pe_config, checkIfExists: true) } // Header files for MultiQC -ch_multiqc_config = file(params.multiqc_config, checkIfExists: true) -ch_mlib_peak_count_header = file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) -ch_mlib_frip_score_header = file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) -ch_mlib_peak_annotation_header = file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) -ch_mlib_deseq2_pca_header = file("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) -ch_mlib_deseq2_clustering_header = file("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) - -ch_mrep_peak_count_header = file("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) -ch_mrep_frip_score_header = file("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) -ch_mrep_peak_annotation_header = file("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) -ch_mrep_deseq2_pca_header = file("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) -ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) +multiqc_config_ch = Channel.fromPath(params.multiqc_config, checkIfExists: true) +mlib_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) +mlib_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) +mlib_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) +mlib_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) +mlib_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) + +mrep_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) +mrep_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) +mrep_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) +mrep_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) +mrep_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) //////////////////////////////////////////////////// /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// // Validate inputs -if (params.design) { ch_design = file(params.design, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } -if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } -if (params.gene_bed) { ch_gene_bed = file(params.gene.bed, checkIfExists: true) } -if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } -if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } +if (params.design){ + design_csv = Channel + .fromPath(params.design, checkIfExists: true) + .ifEmpty { exit 1, "Samples design file not found: ${params.design}" } +} else { + exit 1, "Samples design file not specified!" +} if (params.fasta){ lastPath = params.fasta.lastIndexOf(File.separator) bwa_base = params.fasta.substring(lastPath+1) - ch_fasta = file(params.fasta, checkIfExists: true) + + Channel + .fromPath(params.fasta, checkIfExists: true) + .ifEmpty { exit 1, "Fasta file not found: ${params.fasta}" } + .into { fasta_bwa_index; + fasta_genome_filter; + fasta_markdup_metrics; + fasta_mlib_metrics; + fasta_mlib_macs_annotate; + fasta_mlib_macs_consensus_annotate; + fasta_mrep_macs_annotate; + fasta_mrep_macs_consensus_annotate; + fasta_igv } } else { exit 1, "Fasta file not specified!" } +if (params.gtf){ + Channel + .fromPath(params.gtf, checkIfExists: true) + .ifEmpty { exit 1, "GTF annotation file not found: ${params.gtf}" } + .into { gtf_gene_bed; + gtf_mlib_macs_annotate; + gtf_mlib_macs_consensus_annotate; + gtf_mrep_macs_annotate; + gtf_mrep_macs_consensus_annotate } +} else { + exit 1, "GTF annotation file not specified!" +} + if (params.bwa_index){ lastPath = params.bwa_index.lastIndexOf(File.separator) bwa_dir = params.bwa_index.substring(0,lastPath+1) bwa_base = params.bwa_index.substring(lastPath+1) - ch_bwa_index = Channel + + bwa_index = Channel .fromPath(bwa_dir, checkIfExists: true) .ifEmpty { exit 1, "BWA index directory not found: ${bwa_dir}" } } +if (params.gene_bed){ + gene_bed = Channel + .fromPath(params.gene_bed, checkIfExists: true) + .ifEmpty { exit 1, "Gene BED annotation file not found: ${params.gene_bed}" } +} + +if (params.tss_bed){ + tss_bed = Channel + .fromPath(params.tss_bed, checkIfExists: true) + .ifEmpty { exit 1, "TSS BED annotation file not found: ${params.tss_bed}" } +} + +if (params.blacklist) { + blacklist = Channel + .fromPath(params.blacklist, checkIfExists: true) + .ifEmpty { exit 1, "Blacklist file not found: ${params.blacklist}" } +} + //////////////////////////////////////////////////// /* -- AWS -- */ //////////////////////////////////////////////////// @@ -275,19 +321,18 @@ if (!params.macs_gsize){ /* * PREPROCESSING - REFORMAT DESIGN FILE AND CHECK VALIDITY */ -process check_design { +process reformat_design { tag "$design" input: - file design from ch_design + file design from design_csv output: - stdout into ch_design_bool - file "*.csv" into ch_design_csv + file "*.csv" into reformat_design script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ - check_design.py $design ${design.baseName}.reformat.csv + reformat_design.py $design design_reformat.csv """ } @@ -295,17 +340,34 @@ process check_design { * Create channels for input fastq files */ if (params.singleEnd) { - ch_design_csv.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } - .into { ch_raw_reads_fastqc; - ch_raw_reads_trimgalore } + reformat_design.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } + .into { design_replicates_exist; + design_multiple_samples; + raw_reads_fastqc; + raw_reads_trimgalore } } else { - ch_design_csv.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } - .into { ch_raw_reads_fastqc; - ch_raw_reads_trimgalore } + reformat_design.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } + .into { design_replicates_exist; + design_multiple_samples; + raw_reads_fastqc; + raw_reads_trimgalore } } +// Boolean value for replicates existing in design +replicates_exist = design_replicates_exist.map { it -> it[0].split('_')[-2].replaceAll('R','').toInteger() } + .flatten() + .max() + .val > 1 + +// Boolean value for multiple samples existing in design +multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].join('_') } + .flatten() + .unique() + .count() + .val > 1 + /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// /* -- -- */ @@ -325,10 +387,10 @@ if (!params.bwa_index){ saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' input: - file fasta from ch_fasta + file fasta from fasta_bwa_index output: - file "BWAIndex" into ch_bwa_index + file "BWAIndex" into bwa_index script: """ @@ -347,10 +409,10 @@ if (!params.gene_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file gtf from ch_gtf + file gtf from gtf_gene_bed output: - file "*.bed" into ch_gene_bed + file "*.bed" into gene_bed script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -368,10 +430,10 @@ if (!params.tss_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file bed from ch_gene_bed + file bed from gene_bed output: - file "*.bed" into ch_tss_bed + file "*.bed" into tss_bed script: """ @@ -388,15 +450,15 @@ process makeGenomeFilter { publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file fasta from ch_fasta + file fasta from fasta_genome_filter output: - file "$fasta" into ch_genome_fasta // FASTA FILE FOR IGV - file "*.fai" into ch_genome_fai // FAI INDEX FOR REFERENCE GENOME - file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV - file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING - file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS - ch_genome_sizes_mrep_bigwig + file "$fasta" into genome_fasta // FASTA FILE FOR IGV + file "*.fai" into genome_fai // FAI INDEX FOR REFERENCE GENOME + file "*.txt" into genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV + file "*.bed" into genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING + file "*.sizes" into genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS + genome_sizes_mrep_bigwig script: blacklist_filter = params.blacklist ? "sortBed -i ${params.blacklist} -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" @@ -428,10 +490,10 @@ process fastqc { saveAs: {filename -> filename.endsWith(".zip") ? "zips/$filename" : "$filename"} input: - set val(name), file(reads) from ch_raw_reads_fastqc + set val(name), file(reads) from raw_reads_fastqc output: - file "*.{zip,html}" into ch_fastqc_reports_mqc + file "*.{zip,html}" into fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -462,11 +524,11 @@ process fastqc { * STEP 2 - Trim Galore! */ if (params.skipTrimming){ - ch_trimmed_reads = ch_raw_reads_trimgalore - ch_trimgalore_results_mqc = [] - ch_trimgalore_fastqc_reports_mqc = [] + trimmed_reads = raw_reads_trimgalore + trimgalore_results_mqc = [] + trimgalore_fastqc_reports_mqc = [] } else { - process trimGalore { + process trim_galore { tag "$name" label 'process_long' publishDir "${params.outdir}/trim_galore", mode: 'copy', @@ -478,12 +540,12 @@ if (params.skipTrimming){ } input: - set val(name), file(reads) from ch_raw_reads_trimgalore + set val(name), file(reads) from raw_reads_trimgalore output: - set val(name), file("*.fq.gz") into ch_trimmed_reads - file "*.txt" into ch_trimgalore_results_mqc - file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc + set val(name), file("*.fq.gz") into trimmed_reads + file "*.txt" into trimgalore_results_mqc + file "*.{zip,html}" into trimgalore_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -517,16 +579,16 @@ if (params.skipTrimming){ /* * STEP 3.1 - Align read 1 with bwa */ -process bwaMEM { +process bwa_mem { tag "$name" label 'process_big' input: - set val(name), file(reads) from ch_trimmed_reads - file index from ch_bwa_index.collect() + set val(name), file(reads) from trimmed_reads + file index from bwa_index.first() output: - set val(name), file("*.bam") into ch_bwa_bam + set val(name), file("*.bam") into bwa_bam script: prefix="${name}.Lb" @@ -539,7 +601,7 @@ process bwaMEM { /* * STEP 3.2 - Convert .bam to coordinate sorted .bam */ -process sortBAM { +process sort_bam { tag "$name" label 'process_medium' if (params.saveAlignedIntermediates) { @@ -552,11 +614,11 @@ process sortBAM { } input: - set val(name), file(bam) from ch_bwa_bam + set val(name), file(bam) from bwa_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_mlib - file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into sort_bam_mlib + file "*.{flagstat,idxstats,stats}" into sort_bam_flagstat_mqc script: prefix="${name}.Lb" @@ -580,12 +642,12 @@ process sortBAM { /* * STEP 4.1 Merge BAM files for all libraries from same replicate */ -ch_sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_sort_bam_mlib } +sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { sort_bam_mlib } -process mergeLibBAM { +process merge_library { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -598,13 +660,13 @@ process mergeLibBAM { } input: - set val(name), file(bams) from ch_sort_bam_mlib + set val(name), file(bams) from sort_bam_mlib output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, - ch_mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc - file "*.txt" into ch_mlib_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mlib_bam_filter, + mlib_bam_ataqv + file "*.{flagstat,idxstats,stats}" into mlib_bam_stats_mqc + file "*.txt" into mlib_metrics_mqc script: prefix="${name}.mLb.mkD" @@ -661,7 +723,7 @@ process mergeLibBAM { /* * STEP 4.2 Filter BAM file at merged library-level */ -process mergedLibfilterBAM { +process merge_library_filter { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -676,14 +738,14 @@ process mergedLibfilterBAM { } input: - set val(name), file(bam) from ch_mlib_bam_filter - file bed from ch_genome_filter_regions.collect() - file bamtools_filter_config from ch_bamtools_filter_config + set val(name), file(bam) from mlib_bam_filter + file bed from genome_filter_regions.collect() + file bamtools_filter_config from bamtools_filter_config_ch.collect() output: - set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam - set val(name), file("*.flagstat") into ch_mlib_filter_flagstat - file "*.{idxstats,stats}" into ch_mlib_filter_stats_mqc + set val(name), file("*.{bam,bam.bai}") into mlib_filter_bam + set val(name), file("*.flagstat") into mlib_filter_flagstat + file "*.{idxstats,stats}" into mlib_filter_stats_mqc script: prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" @@ -715,18 +777,18 @@ process mergedLibfilterBAM { * STEP 4.3 Remove orphan reads from paired-end BAM file */ if (params.singleEnd){ - ch_mlib_filter_bam.into { ch_mlib_rm_orphan_bam_metrics; - ch_mlib_rm_orphan_bam_bigwig; - ch_mlib_rm_orphan_bam_macs; - ch_mlib_rm_orphan_bam_mrep; - ch_mlib_name_bam_mlib_counts; - ch_mlib_name_bam_mrep_counts } - ch_mlib_filter_flagstat.into { ch_mlib_rm_orphan_flagstat_bigwig; - ch_mlib_rm_orphan_flagstat_macs; - ch_mlib_rm_orphan_flagstat_mqc } - ch_mlib_filter_stats_mqc.set { ch_mlib_rm_orphan_stats_mqc } + mlib_filter_bam.into { mlib_rm_orphan_bam_metrics; + mlib_rm_orphan_bam_bigwig; + mlib_rm_orphan_bam_macs; + mlib_rm_orphan_bam_mrep; + mlib_name_bam_mlib_counts; + mlib_name_bam_mrep_counts } + mlib_filter_flagstat.into { mlib_rm_orphan_flagstat_bigwig; + mlib_rm_orphan_flagstat_macs; + mlib_rm_orphan_flagstat_mqc } + mlib_filter_stats_mqc.set { mlib_rm_orphan_stats_mqc } } else { - process mergedLibRMOrphanReads { + process merge_library_rm_orphan { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -740,19 +802,19 @@ if (params.singleEnd){ } input: - set val(name), file(bam) from ch_mlib_filter_bam + set val(name), file(bam) from mlib_filter_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, - ch_mlib_rm_orphan_bam_bigwig, - ch_mlib_rm_orphan_bam_macs, - ch_mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, - ch_mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, - ch_mlib_rm_orphan_flagstat_macs, - ch_mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into mlib_rm_orphan_bam_metrics, + mlib_rm_orphan_bam_bigwig, + mlib_rm_orphan_bam_macs, + mlib_rm_orphan_bam_mrep + set val(name), file("${prefix}.bam") into mlib_name_bam_mlib_counts, + mlib_name_bam_mrep_counts + set val(name), file("*.flagstat") into mlib_rm_orphan_flagstat_bigwig, + mlib_rm_orphan_flagstat_macs, + mlib_rm_orphan_flagstat_mqc + file "*.{idxstats,stats}" into mlib_rm_orphan_stats_mqc script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ prefix="${name}.mLb.clN" @@ -779,7 +841,7 @@ if (params.singleEnd){ /* * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries */ -process mergedLibCollectMultipleMetrics { +process merge_library_collectmetrics { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -790,12 +852,12 @@ process mergedLibCollectMultipleMetrics { } input: - set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics - file fasta from ch_fasta + set val(name), file(bam) from mlib_rm_orphan_bam_metrics + file fasta from fasta_mlib_metrics.collect() output: - file "*_metrics" into ch_mlib_collectmetrics_mqc - file "*.pdf" into ch_mlib_collectmetrics_pdf + file "*_metrics" into mlib_collectmetrics_mqc + file "*.pdf" into mlib_collectmetrics_pdf script: prefix="${name}.mLb.clN" @@ -818,7 +880,7 @@ process mergedLibCollectMultipleMetrics { /* * STEP 4.4.2 Read depth normalised bigWig */ -process mergedLibBigWig { +process merge_library_bigwig { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', @@ -829,12 +891,12 @@ process mergedLibBigWig { } input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mlib_bigwig.collect() + set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_bigwig.join(mlib_rm_orphan_flagstat_bigwig, by: [0]) + file sizes from genome_sizes_mlib_bigwig.collect() output: - file "*.bigWig" into ch_mlib_bigwig_igv - file "*.txt" into ch_mlib_bigwig_scale + file "*.bigWig" into mlib_bigwig_igv + file "*.txt" into mlib_bigwig_scale script: prefix="${name}.mLb.clN" @@ -852,7 +914,7 @@ process mergedLibBigWig { /* * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score */ -process mergedLibMACSCallPeak { +process merge_library_macs { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy', @@ -862,18 +924,18 @@ process mergedLibMACSCallPeak { } input: - set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from ch_mlib_peak_count_header - file mlib_frip_score_header from ch_mlib_frip_score_header + set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_macs.join(mlib_rm_orphan_flagstat_macs, by: [0]) + file mlib_peak_count_header from mlib_peak_count_header_ch.collect() + file mlib_frip_score_header from mlib_frip_score_header_ch.collect() output: - set val(name), file("*.{bed,xls,gappedPeak}") into ch_mlib_macs_output - set val(name), file("*$peakext") into ch_mlib_macs_peaks_homer, - ch_mlib_macs_peaks_qc, - ch_mlib_macs_peaks_consensus, - ch_mlib_macs_peaks_ataqv, - ch_mlib_macs_peaks_igv - file "*_mqc.tsv" into ch_mlib_macs_peaks_mqc + set val(name), file("*.{bed,xls,gappedPeak}") into mlib_macs_output + set val(name), file("*$peakext") into mlib_macs_peaks_homer, + mlib_macs_peaks_qc, + mlib_macs_peaks_consensus, + mlib_macs_peaks_ataqv, + mlib_macs_peaks_igv + file "*_mqc.tsv" into mlib_macs_peaks_mqc when: params.macs_gsize @@ -902,17 +964,17 @@ process mergedLibMACSCallPeak { /* * STEP 4.5.2 Annotate peaks with HOMER */ -process mergedLibAnnotatePeaks { +process merge_library_macs_annotate { tag "$name" publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy' input: - set val(name), file(peak) from ch_mlib_macs_peaks_homer - file fasta from ch_fasta - file gtf from ch_gtf + set val(name), file(peak) from mlib_macs_peaks_homer + file fasta from fasta_mlib_macs_annotate.collect() + file gtf from gtf_mlib_macs_annotate.collect() output: - file "*.txt" into ch_mlib_macs_annotate + file "*.txt" into mlib_macs_annotate when: params.macs_gsize @@ -930,18 +992,18 @@ process mergedLibAnnotatePeaks { /* * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process mergedLibPeakQC { +process merge_library_macs_qc { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/qc", mode: 'copy' input: - file peaks from ch_mlib_macs_peaks_qc.collect{ it[1] } - file annos from ch_mlib_macs_annotate.collect() - file mlib_peak_annotation_header from ch_mlib_peak_annotation_header + file peaks from mlib_macs_peaks_qc.collect{ it[1] } + file annos from mlib_macs_annotate.collect() + file mlib_peak_annotation_header from mlib_peak_annotation_header_ch.collect() output: - file "*.{txt,pdf}" into ch_mlib_macs_qc - file "*.tsv" into ch_mlib_macs_qc_mqc + file "*.{txt,pdf}" into mlib_macs_qc + file "*.tsv" into mlib_macs_qc_mqc when: params.macs_gsize @@ -963,30 +1025,23 @@ process mergedLibPeakQC { """ } -// Get boolean values for replicatesExist, multipleGroups, and balancedDesign from checkDesign process -def design_flags(ch) { - ch.map { it.trim().toLowerCase().tokenize() } - .map { it -> [ it[0].asBoolean(), it[1].asBoolean(), it[2].asBoolean() ] } -} - /* * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process mergedLibConsensusPeaks { +process merge_library_macs_consensus { publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: - file peaks from ch_mlib_macs_peaks_consensus.collect{ it[1] } - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file peaks from mlib_macs_peaks_consensus.collect{ it[1] } output: - file "*.bed" into ch_mlib_macs_consensus_bed, - ch_mlib_macs_consensus_bed_igv - file "*.boolean.txt" into ch_mlib_macs_consensus_bool - file "*.saf" into ch_mlib_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect + file "*.bed" into mlib_macs_consensus_bed, + mlib_macs_consensus_bed_igv + file "*.boolean.txt" into mlib_macs_consensus_bool + file "*.saf" into mlib_macs_consensus_saf + file "*.intersect.{txt,plot.pdf}" into mlib_macs_consensus_intersect - when: params.macs_gsize && (replicatesExist || multipleGroups) + when: params.macs_gsize && (multiple_samples || replicates_exist) script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mLb.clN' @@ -1018,20 +1073,19 @@ process mergedLibConsensusPeaks { /* * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process mergedLibConsensusPeaksAnnotate { +process merge_library_macs_consensus_annotate { publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: - file bed from ch_mlib_macs_consensus_bed - file bool from ch_mlib_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file bed from mlib_macs_consensus_bed + file bool from mlib_macs_consensus_bool + file fasta from fasta_mlib_macs_consensus_annotate + file gtf from gtf_mlib_macs_consensus_annotate output: file "*.annotatePeaks.txt" into mlib_macs_consensus_annotate - when: params.macs_gsize && (replicatesExist || multipleGroups) + when: params.macs_gsize && (multiple_samples || replicates_exist) script: prefix="consensus_peaks.mLb.clN" @@ -1050,27 +1104,26 @@ process mergedLibConsensusPeaksAnnotate { /* * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process mergedLibConsensusPeaksDESeq { +process merge_library_macs_consensus_deseq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus/deseq2", mode: 'copy' input: - file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from ch_mlib_macs_consensus_saf.collect() - file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header - file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file bams from mlib_name_bam_mlib_counts.collect{ it[1] } + file saf from mlib_macs_consensus_saf.collect() + file mlib_deseq2_pca_header from mlib_deseq2_pca_header_ch.collect() + file mlib_deseq2_clustering_header from mlib_deseq2_clustering_header_ch.collect() output: - file "*featureCounts.txt" into ch_mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results - file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc + file "*featureCounts.txt" into mlib_macs_consensus_counts + file "*featureCounts.txt.summary" into mlib_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into mlib_macs_consensus_deseq_results + file "sizeFactors" into mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into mlib_macs_consensus_deseq_comp_bed_igv + file "*.tsv" into mlib_macs_consensus_deseq_mqc - when: params.macs_gsize && replicatesExist && multipleGroups + when: params.macs_gsize && multiple_samples && replicates_exist script: prefix="consensus_peaks.mLb.clN" @@ -1097,18 +1150,18 @@ process mergedLibConsensusPeaksDESeq { /* * STEP 4.6.1 Run ataqv on BAM file and corresponding peaks */ -process mergedLibAtaqv { +process merge_library_ataqv { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' input: - set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_peaks_ataqv, by: [0]) - file tss_bed from ch_tss_bed.collect() - file autosomes from ch_genome_autosomes.collect() + set val(name), file(bam), file(peak) from mlib_bam_ataqv.join(mlib_macs_peaks_ataqv, by: [0]) + file tss_bed from tss_bed.collect() + file autosomes from genome_autosomes.collect() output: - file "*.json" into ch_mlib_ataqv + file "*.json" into mlib_ataqv script: peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" @@ -1130,15 +1183,15 @@ process mergedLibAtaqv { /* * STEP 4.6.2 run ataqv mkarv on all JSON files to render web app */ -process mergedLibMkarv { +process merge_library_ataqv_mkarv { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' input: - file json from ch_mlib_ataqv.collect() + file json from mlib_ataqv.collect() output: - file "html" into ch_mlib_ataqv_mkarv + file "html" into mlib_ataqv_mkarv script: """ @@ -1160,12 +1213,12 @@ process mergedLibMkarv { /* * STEP 5.1 Merge library BAM files across all replicates */ -ch_mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { ch_mlib_rm_orphan_bam_mrep } +mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { mlib_rm_orphan_bam_mrep } -process mergeRepBAM { +process merge_replicate { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', @@ -1178,19 +1231,18 @@ process mergeRepBAM { } input: - set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + set val(name), file(bams) from mlib_rm_orphan_bam_mrep output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, - ch_mrep_bam_macs - set val(name), file("*.flagstat") into ch_mrep_flagstat_bigwig, - ch_mrep_flagstat_macs, - ch_mrep_flagstat_mqc - file "*.{idxstats,stats}" into ch_mrep_stats_mqc - file "*.txt" into ch_mrep_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mrep_bam_bigwig, + mrep_bam_macs + set val(name), file("*.flagstat") into mrep_flagstat_bigwig, + mrep_flagstat_macs, + mrep_flagstat_mqc + file "*.{idxstats,stats}" into mrep_stats_mqc + file "*.txt" into mrep_metrics_mqc - when: !params.skipMergeReplicates && replicatesExist + when: !params.skipMergeReplicates && replicates_exist script: prefix="${name}.mRp.clN" @@ -1248,7 +1300,7 @@ process mergeRepBAM { /* * STEP 5.2.1 Read depth normalised bigWig */ -process mergedRepBigWig { +process merge_replicate_bigwig { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', @@ -1259,15 +1311,14 @@ process mergedRepBigWig { } input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_flagstat_bigwig, by: [0]) - file sizes from ch_genome_sizes_mrep_bigwig.collect() - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + set val(name), file(bam), file(flagstat) from mrep_bam_bigwig.join(mrep_flagstat_bigwig, by: [0]) + file sizes from genome_sizes_mrep_bigwig.collect() output: - file "*.bigWig" into ch_mrep_bigwig_igv - file "*.txt" into ch_mrep_bigwig_scale + file "*.bigWig" into mrep_bigwig_igv + file "*.txt" into mrep_bigwig_scale - when: !params.skipMergeReplicates && replicatesExist + when: !params.skipMergeReplicates && replicates_exist script: prefix="${name}.mRp.clN" @@ -1285,7 +1336,7 @@ process mergedRepBigWig { /* * STEP 5.2.2 Call peaks with MACS2 and calculate FRiP score */ -process mergedRepMACSCallPeak { +process merge_replicate_macs { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy', @@ -1295,20 +1346,19 @@ process mergedRepMACSCallPeak { } input: - set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_flagstat_macs, by: [0]) - file mrep_peak_count_header from ch_mrep_peak_count_header - file mrep_frip_score_header from ch_mrep_frip_score_header - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + set val(name), file(bam), file(flagstat) from mrep_bam_macs.join(mrep_flagstat_macs, by: [0]) + file mrep_peak_count_header from mrep_peak_count_header_ch.collect() + file mrep_frip_score_header from mrep_frip_score_header_ch.collect() output: - file "*.{bed,xls,gappedPeak}" into ch_mrep_macs_output - set val(name), file("*$peakext") into ch_mrep_macs_peaks_homer, - ch_mrep_macs_peaks_qc, - ch_mrep_macs_peaks_consensus, - ch_mrep_macs_peaks_igv - file "*_mqc.tsv" into ch_mrep_macs_peak_mqc + file "*.{bed,xls,gappedPeak}" into mrep_macs_output + set val(name), file("*$peakext") into mrep_macs_peaks_homer, + mrep_macs_peaks_qc, + mrep_macs_peaks_consensus, + mrep_macs_peaks_igv + file "*_mqc.tsv" into mrep_macs_peak_mqc - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: prefix="${name}.mRp.clN" @@ -1335,20 +1385,19 @@ process mergedRepMACSCallPeak { /* * STEP 5.2.3 Annotate peaks with HOMER */ -process mergedRepAnnotatePeaks { +process merge_replicate_macs_annotate { tag "$name" publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy' input: - set val(name), file(peak) from ch_mrep_macs_peaks_homer - file fasta from ch_fasta - file gtf from ch_gtf - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + set val(name), file(peak) from mrep_macs_peaks_homer + file fasta from fasta_mrep_macs_annotate.collect() + file gtf from gtf_mrep_macs_annotate.collect() output: - file "*.txt" into ch_mrep_macs_annotate + file "*.txt" into mrep_macs_annotate - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: prefix="${name}.mRp.clN" @@ -1364,21 +1413,20 @@ process mergedRepAnnotatePeaks { /* * STEP 5.2.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process mergedRepPeakQC { +process merge_replicate_macs_qc { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/qc", mode: 'copy' input: - file peaks from ch_mrep_macs_peaks_qc.collect{ it[1] } - file annos from ch_mrep_macs_annotate.collect() - file mrep_peak_annotation_header from ch_mrep_peak_annotation_header - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file peaks from mrep_macs_peaks_qc.collect{ it[1] } + file annos from mrep_macs_annotate.collect() + file mrep_peak_annotation_header from mrep_peak_annotation_header_ch.collect() output: - file "*.{txt,pdf}" into ch_mrep_macs_qc - file "*.tsv" into ch_mrep_macs_qc_mqc + file "*.{txt,pdf}" into mrep_macs_qc + file "*.tsv" into mrep_macs_qc_mqc - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mRp.clN' @@ -1401,21 +1449,20 @@ process mergedRepPeakQC { /* * STEP 5.2.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process mergedRepConsensusPeaks { +process merge_replicate_macs_consensus { publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: - file peaks from ch_mrep_macs_peaks_consensus.collect{ it[1] } - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file peaks from mrep_macs_peaks_consensus.collect{ it[1] } output: - file "*.bed" into ch_mrep_macs_consensus_bed, - ch_mrep_macs_consensus_bed_igv - file "*.boolean.txt" into ch_mrep_macs_consensus_bool - file "*.saf" into ch_mrep_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect + file "*.bed" into mrep_macs_consensus_bed, + mrep_macs_consensus_bed_igv + file "*.boolean.txt" into mrep_macs_consensus_bool + file "*.saf" into mrep_macs_consensus_saf + file "*.intersect.{txt,plot.pdf}" into mrep_macs_consensus_intersect - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ suffix='mRp.clN' @@ -1447,20 +1494,19 @@ process mergedRepConsensusPeaks { /* * STEP 5.2.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process mergedRepConsensusPeaksAnnotate { +process merge_replicate_macs_consensus_annotate { publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: - file bed from ch_mrep_macs_consensus_bed - file bool from ch_mrep_macs_consensus_bool - file fasta from ch_fasta - file gtf from ch_gtf - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file bed from mrep_macs_consensus_bed + file bool from mrep_macs_consensus_bool + file fasta from fasta_mrep_macs_consensus_annotate + file gtf from gtf_mrep_macs_consensus_annotate output: - file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate + file "*.annotatePeaks.txt" into mrep_macs_consensus_annotate - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: prefix="consensus_peaks.mRp.clN" @@ -1479,27 +1525,26 @@ process mergedRepConsensusPeaksAnnotate { /* * STEP 5.2.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process mergedRepConsensusPeaksDESeq { +process merge_replicate_macs_consensus_deseq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus/deseq2", mode: 'copy' input: - file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from ch_mrep_macs_consensus_saf.collect() - file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header - file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header - set val(replicatesExist), val(multipleGroups), val(balancedDesign) from design_flags(ch_design_bool) + file bams from mlib_name_bam_mrep_counts.collect{ it[1] } + file saf from mrep_macs_consensus_saf.collect() + file mrep_deseq2_pca_header from mrep_deseq2_pca_header_ch.collect() + file mrep_deseq2_clustering_header from mrep_deseq2_clustering_header_ch.collect() output: - file "*featureCounts.txt" into ch_mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results - file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc + file "*featureCounts.txt" into mrep_macs_consensus_counts + file "*featureCounts.txt.summary" into mrep_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into mrep_macs_consensus_deseq_results + file "sizeFactors" into mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into mrep_macs_consensus_deseq_comp_bed_igv + file "*.tsv" into mrep_macs_consensus_deseq_mqc - when: !params.skipMergeReplicates && params.macs_gsize && replicatesExist && multipleGroups + when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: prefix="consensus_peaks.mRp.clN" @@ -1538,20 +1583,20 @@ process igv { publishDir "${params.outdir}/igv", mode: 'copy' input: - file fasta from ch_fasta + file fasta from fasta_igv.collect() - file ('bwa/mergedLibrary/bigwig/*') from ch_mlib_bigwig_igv.collect() - file ('bwa/mergedLibrary/macs/*') from ch_mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/*') from ch_mlib_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/deseq2/*') from ch_mlib_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedLibrary/bigwig/*') from mlib_bigwig_igv.collect() + file ('bwa/mergedLibrary/macs/*') from mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) + file ('bwa/mergedLibrary/macs/consensus/*') from mlib_macs_consensus_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedLibrary/macs/consensus/deseq2/*') from mlib_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/bigwig/*') from ch_mrep_bigwig_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/*') from ch_mrep_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/*') from ch_mrep_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/deseq2/*') from ch_mrep_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/bigwig/*') from mrep_bigwig_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/macs/*') from mrep_macs_peaks_igv.collect{it[1]}.ifEmpty([]) + file ('bwa/mergedReplicate/macs/consensus/*') from mrep_macs_consensus_bed_igv.collect().ifEmpty([]) + file ('bwa/mergedReplicate/macs/consensus/deseq2/*') from mrep_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) output: - file "*.{txt,xml}" into ch_igv_session + file "*.{txt,xml}" into igv_session script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ outdir_abspath = new File(params.outdir).getCanonicalPath().toString() @@ -1581,7 +1626,7 @@ process get_software_versions { } output: - file 'software_versions_mqc.yaml' into ch_software_versions_mqc + file 'software_versions_mqc.yaml' into software_versions_mqc file "software_versions.csv" script: @@ -1631,37 +1676,37 @@ process multiqc { publishDir "${params.outdir}/multiqc", mode: 'copy' input: - file multiqc_config from ch_multiqc_config - - file ('fastqc/*') from ch_fastqc_reports_mqc.collect() - file ('trimgalore/*') from ch_trimgalore_results_mqc.collect() - file ('trimgalore/fastqc/*') from ch_trimgalore_fastqc_reports_mqc.collect() - - file ('alignment/library/*') from ch_sort_bam_flagstat_mqc.collect() - - file ('alignment/mergedLibrary/*') from ch_mlib_bam_stats_mqc.collect() - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_flagstat_mqc.collect{it[1]} - file ('alignment/mergedLibrary/*') from ch_mlib_rm_orphan_stats_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_metrics_mqc.collect() - file ('alignment/mergedLibrary/picard_metrics/*') from ch_mlib_collectmetrics_mqc.collect() - file ('macs/mergedLibrary/*') from ch_mlib_macs_peaks_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/*') from ch_mlib_macs_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedLibrary/consensus/*') from ch_mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - file ('alignment/mergedReplicate/*') from ch_mrep_flagstat_mqc.collect{it[1]}.ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_stats_mqc.collect().ifEmpty([]) - file ('alignment/mergedReplicate/*') from ch_mrep_metrics_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_macs_peak_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/*') from ch_mrep_macs_qc_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) - file ('macs/mergedReplicate/consensus/*') from ch_mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) - - file ('software_versions/*') from ch_software_versions_mqc.collect() + file multiqc_config from multiqc_config_ch.collect() + + file ('fastqc/*') from fastqc_reports_mqc.collect() + file ('trimgalore/*') from trimgalore_results_mqc.collect() + file ('trimgalore/fastqc/*') from trimgalore_fastqc_reports_mqc.collect() + + file ('alignment/library/*') from sort_bam_flagstat_mqc.collect() + + file ('alignment/mergedLibrary/*') from mlib_bam_stats_mqc.collect() + file ('alignment/mergedLibrary/*') from mlib_rm_orphan_flagstat_mqc.collect{it[1]} + file ('alignment/mergedLibrary/*') from mlib_rm_orphan_stats_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from mlib_metrics_mqc.collect() + file ('alignment/mergedLibrary/picard_metrics/*') from mlib_collectmetrics_mqc.collect() + file ('macs/mergedLibrary/*') from mlib_macs_peaks_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/*') from mlib_macs_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from mlib_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedLibrary/consensus/*') from mlib_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + file ('alignment/mergedReplicate/*') from mrep_flagstat_mqc.collect{it[1]}.ifEmpty([]) + file ('alignment/mergedReplicate/*') from mrep_stats_mqc.collect().ifEmpty([]) + file ('alignment/mergedReplicate/*') from mrep_metrics_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from mrep_macs_peak_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/*') from mrep_macs_qc_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from mrep_macs_consensus_counts_mqc.collect().ifEmpty([]) + file ('macs/mergedReplicate/consensus/*') from mrep_macs_consensus_deseq_mqc.collect().ifEmpty([]) + + file ('software_versions/*') from software_versions_mqc.collect() file ('workflow_summary/*') from create_workflow_summary(summary) output: - file "*multiqc_report.html" into ch_multiqc_report + file "*multiqc_report.html" into multiqc_report file "*_data" file "multiqc_plots" @@ -1689,7 +1734,7 @@ process output_documentation { publishDir "${params.outdir}/Documentation", mode: 'copy' input: - file output_docs from ch_output_docs + file output_docs from output_docs_ch output: file "results_description.html" From a978b4f61befd93547534a983e03eff0c4f877d1 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:17:06 +0100 Subject: [PATCH 013/149] Update README --- README.md | 10 +++++----- 1 file changed, 5 insertions(+), 5 deletions(-) diff --git a/README.md b/README.md index 9c853408..a1c2d25f 100644 --- a/README.md +++ b/README.md @@ -7,13 +7,13 @@ [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) -### Introduction +## Introduction **nfcore/atacseq** is a bioinformatics analysis pipeline used for ATAC-seq data. The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool to run tasks across multiple compute infrastructures in a very portable manner. It comes with docker containers making installation trivial and results highly reproducible. -### Pipeline summary +## Pipeline summary 1. Raw read QC ([`FastQC`](https://www.bioinformatics.babraham.ac.uk/projects/fastqc/)) 2. Adapter trimming ([`Trim Galore!`](https://www.bioinformatics.babraham.ac.uk/projects/trim_galore/)) @@ -53,7 +53,7 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool 7. Create IGV session file containing bigWig tracks, peaks and differential sites for data visualisation ([`IGV`](https://software.broadinstitute.org/software/igv/)). 8. Present QC for raw read, alignment, peak-calling and differential accessibility results ([`ataqv`](https://github.com/ParkerLab/ataqv), [`MultiQC`](http://multiqc.info/), [`R`](https://www.r-project.org/)) -### Documentation +## Documentation The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: 1. [Installation](https://nf-co.re/usage/installation) @@ -65,9 +65,9 @@ The nf-core/atacseq pipeline comes with documentation about the pipeline, found 4. [Output and how to interpret the results](docs/output.md) 5. [Troubleshooting](https://nf-co.re/usage/troubleshooting) -### Credits +## Credits -The pipeline was originally written by the [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. +The pipeline was originally written by [The Bioinformatics & Biostatistics Group](https://www.crick.ac.uk/research/science-technology-platforms/bioinformatics-and-biostatistics/) for use at [The Francis Crick Institute](https://www.crick.ac.uk/), London. The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). From 1dc55c7d801a6ffc641568d4798909fbda1fe576 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:19:06 +0100 Subject: [PATCH 014/149] Add Quick Start info --- README.md | 20 ++++++++++++++++++++ 1 file changed, 20 insertions(+) diff --git a/README.md b/README.md index a1c2d25f..15a8b962 100644 --- a/README.md +++ b/README.md @@ -53,6 +53,26 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool 7. Create IGV session file containing bigWig tracks, peaks and differential sites for data visualisation ([`IGV`](https://software.broadinstitute.org/software/igv/)). 8. Present QC for raw read, alignment, peak-calling and differential accessibility results ([`ataqv`](https://github.com/ParkerLab/ataqv), [`MultiQC`](http://multiqc.info/), [`R`](https://www.r-project.org/)) +## Quick Start + +i. Install [`nextflow`](https://nf-co.re/usage/installation) + +ii. Install one of [`docker`](https://docs.docker.com/engine/installation/), [`singularity`](https://www.sylabs.io/guides/3.0/user-guide/) or [`conda`](https://conda.io/miniconda.html) + +iii. Download the pipeline and test it on a minimal dataset with a single command + +```bash +nextflow run nf-core/atacseq -profile test, +``` + +iv. Start running your own analysis! + +```bash +nextflow run nf-core/atacseq -profile --design design.csv --genome GRCh37 +``` + +See [usage docs](docs/usage.md) for all of the available options when running the pipeline. + ## Documentation The nf-core/atacseq pipeline comes with documentation about the pipeline, found in the `docs/` directory: From 566d4dab191a4909afdf830dea8c60ebeaa5befc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:20:33 +0100 Subject: [PATCH 015/149] Remove fastq file existence check --- bin/reformat_design.py | 10 ---------- 1 file changed, 10 deletions(-) diff --git a/bin/reformat_design.py b/bin/reformat_design.py index cf7aafa2..8647b0c7 100644 --- a/bin/reformat_design.py +++ b/bin/reformat_design.py @@ -76,16 +76,6 @@ def reformat_design(DesignFileIn,DesignFileOut): print "{}: FastQ file has incorrect extension (has to be '.fastq.gz' or 'fq.gz') - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) sys.exit(1) - ## CHECK FASTQ FILES EXIST PER SAMPLE - if fastq[:4] not in ['http']: - if not os.path.exists(fastq): - print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) - sys.exit(1) - else: - if requests.head(fastq).status_code >= 400: - print "{}: FastQ file does not exist - {}\nLine: '{}'".format(ERROR_STR,fastq,line.strip()) - sys.exit(1) - ## CREATE GROUP MAPPING DICT = {GROUP_ID: {REPLICATE_ID:[[FASTQ_FILES]]} replicate = int(replicate) if not groupRepDict.has_key(group): From c4adc20aaa0476c3b4d1b65281ea267ab75808ab Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:31:55 +0100 Subject: [PATCH 016/149] Add bed12 paths --- conf/igenomes.config | 26 ++++++++++++++++++++++++++ 1 file changed, 26 insertions(+) diff --git a/conf/igenomes.config b/conf/igenomes.config index e8c591fa..3c36daa2 100755 --- a/conf/igenomes.config +++ b/conf/igenomes.config @@ -14,6 +14,7 @@ params { fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/GRCh37-blacklist.bed" macs_gsize = "2.7e9" mito_name = "MT" @@ -22,6 +23,7 @@ params { fasta = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" macs_gsize = "2.7e9" mito_name = "chrM" @@ -30,6 +32,7 @@ params { fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/GRCm38-blacklist.bed" macs_gsize = "1.87e9" mito_name = "MT" @@ -38,23 +41,27 @@ params { fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" mito_name = "Mt" } 'EB2' { fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" } 'UMD3.1' { fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" mito_name = "MT" } 'WBcel235' { fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" macs_gsize = "9e7" mito_name = "MtDNA" } @@ -62,18 +69,21 @@ params { fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" mito_name = "MT" } 'GRCz10' { fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" mito_name = "MT" } 'BDGP6' { fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" macs_gsize = "1.2e8" mito_name = "M" } @@ -81,52 +91,61 @@ params { fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" mito_name = "MT" } 'EB1' { fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" } 'Galgal4' { fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" mito_name = "MT" } 'Gm01' { fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" } 'Mmul_1' { fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" mito_name = "MT" } 'IRGSP-1.0' { fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" mito_name = "Mt" } 'CHIMP2.1.4' { fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" mito_name = "MT" } 'Rnor_6.0' { fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" mito_name = "MT" } 'R64-1-1' { fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" macs_gsize = "1.2e7" mito_name = "MT" } @@ -134,6 +153,7 @@ params { fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" macs_gsize = "1.21e7" mito_name = "MT" } @@ -141,23 +161,27 @@ params { fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" } 'Sscrofa10.2' { fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" mito_name = "MT" } 'AGPv3' { fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" mito_name = "Mt" } 'hg38' { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" macs_gsize = "2.7e9" mito_name = "chrM" @@ -166,6 +190,7 @@ params { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/hg19-blacklist.bed" macs_gsize = "2.7e9" mito_name = "chrM" @@ -174,6 +199,7 @@ params { fasta = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/genome.fa" gtf = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed" blacklist = "${baseDir}/assets/blacklists/mm10-blacklist.bed" macs_gsize = "1.87e9" mito_name = "chrM" From 2f2b6f97694ca9ef6adc9daeddd565644c00a8fc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:37:10 +0100 Subject: [PATCH 017/149] Render example plots inline --- docs/output.md | 47 ++++++++++++++++++++--------------------------- 1 file changed, 20 insertions(+), 27 deletions(-) diff --git a/docs/output.md b/docs/output.md index ae530a0e..ef0f710e 100755 --- a/docs/output.md +++ b/docs/output.md @@ -45,8 +45,7 @@ The initial QC and alignments are performed at the library-level e.g. if the sam * `trim_galore/fastqc/zips/` FastQC `*.zip` files for read 1 (*and read2 if paired-end*) **after** adapter trimming. - *Plots*: - [MultiQC - Cutadapt trimmed sequence plot](images/mqc_cutadapt_plot.png) + ![MultiQC - Cutadapt trimmed sequence plot](images/mqc_cutadapt_plot.png) 3. **Alignment** @@ -64,8 +63,7 @@ The initial QC and alignments are performed at the library-level e.g. if the sam * `bwa/library/samtools_stats/` SAMtools `*.flagstat`, `*.idxstats` and `*.stats` files generated from the alignment files. - *Plots*: - [MultiQC - SAMtools idxstats plot](images/mqc_samtools_idxstats_plot.png) + ![MultiQC - SAMtools idxstats plot](images/mqc_samtools_idxstats_plot.png) ## Merged library-level analysis @@ -97,9 +95,8 @@ The library-level alignments associated with the same sample are merged and subs * `bwa/mergedLibrary/picard_metrics/pdf/` Alignment QC plot files in `*.pdf` format from picard CollectMultipleMetrics. - *Plots*: - [MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) - [MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) + ![MultiQC - Picard insert size plot](images/mqc_picard_insert_size_plot.png) + ![MultiQC - Picard deduplication stats plot](images/mqc_picard_deduplication_plot.png) 2. **Normalised bigWig files** @@ -130,19 +127,18 @@ The library-level alignments associated with the same sample are merged and subs See [MACS2 outputs](https://github.com/taoliu/MACS#output-files) for a description of the output files generated by MACS2. *Output directories*: - * `bwa/mergedLibrary/macs2/` + * `bwa/mergedLibrary/macs/` * MACS2 output files: `*.xls`, `*.broadPeak` or `*.narrowPeak`, `*.gappedPeak` and `*summits.bed`. The files generated will depend on whether MACS2 has been run in *narrowPeak* or *broadPeak* mode. * HOMER peak-to-gene annotation file: `*.annotatePeaks.txt`. - * `bwa/mergedLibrary/macs2/qc` + * `bwa/mergedLibrary/macs/qc` * QC plots for MACS2 peaks: `macs_peak.mLb.clN.plots.pdf` * QC plots for peak-to-gene feature annotation: `macs_annotatePeaks.mLb.clN.plots.pdf` * MultiQC custom-content files for [FRiP score](https://genome.cshlp.org/content/22/9/1813.full.pdf+html), peak count and peak-to-gene ratios: `*.FRiP_mqc.tsv`, `*.count_mqc.tsv` and `macs_annotatePeaks.mLb.clN.summary_mqc.tsv` respectively. - *Plots*: - [MultiQC - MACS2 total peak count plot](images/mqc_macs2_peak_count_plot.png) - [MultiQC - MACS2 peaks FRiP score plot](images/mqc_frip_score_plot.png) - [MultiQC - HOMER annotatePeaks peak-to-gene feature ratio plot](images/mqc_annotatePeaks_feature_percentage_plot.png) + ![MultiQC - MACS2 total peak count plot](images/mqc_macs2_peak_count_plot.png) + ![MultiQC - MACS2 peaks FRiP score plot](images/mqc_frip_score_plot.png) + ![MultiQC - HOMER annotatePeaks peak-to-gene feature ratio plot](images/mqc_annotatePeaks_feature_percentage_plot.png) 4. **Create consensus set of peaks** @@ -155,7 +151,7 @@ The library-level alignments associated with the same sample are merged and subs Using the consensus peaks it is also possible to assess the degree of overlap between the peaks from a set of samples e.g. *Which consensus peaks contain peaks that are common to a given set of samples?*. This may be useful for downstream filtering of peaks based on whether they are called in multiple replicates/conditions. Please note that it is possible for a consensus peak to contain multiple peaks from the same sample. Unfortunately, this is sample-dependent but the files generated by the pipeline do have columns that report such instances and allow you to factor them into any further analysis. *Output directories*: - * `bwa/mergedLibrary/macs2/consensus/` + * `bwa/mergedLibrary/macs/consensus/` * Consensus peakset across all samples in `*.bed` format. * Consensus peakset across all samples in `*.saf` format. Required by featureCounts for read quantification. * HOMER `*.annotatePeaks.txt` peak-to-gene annotation file for consensus peaks. @@ -165,8 +161,7 @@ The library-level alignments associated with the same sample are merged and subs Same as file above but without annotation columns. * [UpSetR](https://cran.r-project.org/web/packages/UpSetR/README.html) files to illustrate peak intersection: `*.boolean.intersect.plot.pdf` and `*.boolean.intersect.txt`. - *Plots*: - [R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) + ![R - UpSetR peak intersection plot](images/r_upsetr_intersect_plot.png) 5. **Read counting and differential accessibility analysis** @@ -185,27 +180,26 @@ The library-level alignments associated with the same sample are merged and subs Please see [DESeq2 output](http://bioconductor.org/packages/release/bioc/vignettes/DESeq2/inst/doc/DESeq2.html#differential-expression-analysis) for a description of the columns generated by DESeq2. *Output directories*: - * `bwa/mergedLibrary/macs2/consensus/deseq2/` + * `bwa/mergedLibrary/macs/consensus/deseq2/` * `.featureCounts.txt` file for read counts across all samples relative to consensus peak set. * Differential accessibility `*.results.txt` spreadsheet containing results across all consensus peaks and all comparisons. * `*.plots.pdf` file for PCA and hierarchical clustering. * `*.log` file with information for number of differentially bound intervals at different FDR and fold-change thresholds for each comparison. * `*.dds.rld.RData` file containing R `dds` and `rld` objects generated by DESeq2. * `R_sessionInfo.log` file containing information about R, the OS and attached or loaded packages. - * `bwa/mergedLibrary/macs2/consensus//` + * `bwa/mergedLibrary/macs/consensus//` * `*.results.txt` spreadsheet containing comparison-specific DESeq2 output for differential accessibility results across all peaks. * Subset of above file for peaks that pass FDR <= 0.01 (`*FDR0.01.results.txt`) and FDR <= 0.05 (`*FDR0.05.results.txt`). * BED files for peaks that pass FDR <= 0.01 (`*FDR0.01.results.bed`) and FDR <= 0.05 (`*FDR0.05.results.bed`). * MA, Volcano, clustering and scatterplots at FDR <= 0.01 and FDR <= 0.05: `*deseq2.plots.pdf`. - * `bwa/mergedLibrary/macs2/consensus/sizeFactors/` + * `bwa/mergedLibrary/macs/consensus/sizeFactors/` Files containing DESeq2 sizeFactors per sample: `*.txt` and `*.RData`. - *Plots*: - [MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) - [MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) - [MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) - [R - DESeq2 MA plot](images/r_deseq2_ma_plot.png) - [R - DESeq2 Volcano plot](images/r_deseq2_volcano_plot.png) + ![MultiQC - featureCounts consensus peak read assignment plot](images/mqc_featureCounts_assignment_plot.png) + ![MultiQC - DESeq2 PCA plot](images/mqc_deseq2_pca_plot.png) + ![MultiQC - DESeq2 sample similarity plot](images/mqc_deseq2_sample_similarity_plot.png) + ![R - DESeq2 MA plot](images/r_deseq2_ma_plot.png) + ![R - DESeq2 Volcano plot](images/r_deseq2_volcano_plot.png) 6. **ataqv** @@ -276,8 +270,7 @@ You can skip this portion of the analysis by specifying the `--skipMergeReplicat * `igv_session.xml` file. * `igv_files.txt` file containing a listing of the files used to create the IGV session, and their associated colours. - *Plots*: - [IGV screenshot](images/igv_screenshot.png) + ![IGV screenshot](images/igv_screenshot.png) ## Other results From bce8e60ab9ae3f2213a4e68d47bd36964e44aeaf Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:39:46 +0100 Subject: [PATCH 018/149] Evaluate bed12 from igenomes.config --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 7fc64584..3ec4cf54 100644 --- a/main.nf +++ b/main.nf @@ -107,7 +107,7 @@ if (params.genomes && params.genome && !params.genomes.containsKey(params.genome params.fasta = params.genome ? params.genomes[ params.genome ].fasta ?: false : false params.bwa_index = params.genome ? params.genomes[ params.genome ].bwa ?: false : false params.gtf = params.genome ? params.genomes[ params.genome ].gtf ?: false : false -params.gene_bed = params.genome ? params.genomes[ params.genome ].gene_bed ?: false : false +params.gene_bed = params.genome ? params.genomes[ params.genome ].bed12 ?: false : false params.mito_name = params.genome ? params.genomes[ params.genome ].mito_name ?: false : false params.macs_gsize = params.genome ? params.genomes[ params.genome ].macs_gsize ?: false : false params.blacklist = params.genome ? params.genomes[ params.genome ].blacklist ?: false : false From 16768e47e7165be950428837e2572ecea8e13447 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 21:57:18 +0100 Subject: [PATCH 019/149] Update CHANGELOG --- CHANGELOG.md | 8 ++++++++ 1 file changed, 8 insertions(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index c5744b4f..0e632f48 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,6 +10,14 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * Merged in TEMPLATE branch for automated syncing +* Add quick start information to main README +* [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config + +### `Fixed` + +* [#41](https://github.com/nf-core/atacseq/issues/41) - Docs: Add example plot images +* [#44](https://github.com/nf-core/atacseq/issues/44) - Output directory missing: macs2/consensus/deseq2 +* Smaller logo for completion email ## [1.0.0] - 2019-04-09 From afa1b30ff6fae67ca0948ab2e0ce67c82e8a6139 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 22:26:45 +0100 Subject: [PATCH 020/149] Change file permissions --- bin/reformat_design.py | 0 1 file changed, 0 insertions(+), 0 deletions(-) mode change 100644 => 100755 bin/reformat_design.py diff --git a/bin/reformat_design.py b/bin/reformat_design.py old mode 100644 new mode 100755 From 963ba7f92c0fd1632c1672504520cdbec9dcad41 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Sun, 29 Sep 2019 22:42:29 +0100 Subject: [PATCH 021/149] Remove Fmt --- main.nf | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/main.nf b/main.nf index 3ec4cf54..7a7e70a6 100644 --- a/main.nf +++ b/main.nf @@ -1839,10 +1839,10 @@ workflow.onComplete { c_green = params.monochrome_logs ? '' : "\033[0;32m"; c_red = params.monochrome_logs ? '' : "\033[0;31m"; - if (workflow.stats.ignoredCountFmt > 0 && workflow.success) { + if (workflow.stats.ignoredCount > 0 && workflow.success) { log.info "${c_purple}Warning, pipeline completed, but with errored process(es) ${c_reset}" - log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCountFmt} ${c_reset}" - log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCountFmt} ${c_reset}" + log.info "${c_red}Number of ignored errored process(es) : ${workflow.stats.ignoredCount} ${c_reset}" + log.info "${c_green}Number of successfully ran process(es) : ${workflow.stats.succeedCount} ${c_reset}" } if(workflow.success){ From a589ea88e6655d50b697ab3c0c45b35e9604a43a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 17:19:18 +0100 Subject: [PATCH 022/149] Bump requirements for annotatePeaks --- CHANGELOG.md | 1 + main.nf | 6 +++++- 2 files changed, 6 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 0e632f48..df0e6425 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -12,6 +12,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Merged in TEMPLATE branch for automated syncing * Add quick start information to main README * [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config +* [#46](https://github.com/nf-core/atacseq/issues/46) - Bump resources for MAC2 and annotatePeaks.pl ### `Fixed` diff --git a/main.nf b/main.nf index c6559483..c148c292 100644 --- a/main.nf +++ b/main.nf @@ -464,7 +464,7 @@ process makeGenomeFilter { genome_sizes_mrep_bigwig script: - blacklist_filter = params.blacklist ? "sortBed -i ${blacklist} -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" + blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" mito_filter = params.keepMito ? "" : name_filter """ @@ -969,6 +969,7 @@ process merge_library_macs { */ process merge_library_macs_annotate { tag "$name" + label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy' input: @@ -1077,6 +1078,7 @@ process merge_library_macs_consensus { * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process merge_library_macs_consensus_annotate { + label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: @@ -1390,6 +1392,7 @@ process merge_replicate_macs { */ process merge_replicate_macs_annotate { tag "$name" + label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy' input: @@ -1498,6 +1501,7 @@ process merge_replicate_macs_consensus { * STEP 5.2.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process merge_replicate_macs_consensus_annotate { + label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: From 22407550238e5db5c45e711f9a3bc7bff6ffc954 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 21:52:47 +0100 Subject: [PATCH 023/149] Rename input file channels --- CHANGELOG.md | 1 + main.nf | 169 ++++++++++++++++++--------------------------------- 2 files changed, 61 insertions(+), 109 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index df0e6425..b8c0177a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -13,6 +13,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * Add quick start information to main README * [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config * [#46](https://github.com/nf-core/atacseq/issues/46) - Bump resources for MAC2 and annotatePeaks.pl +* [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly ### `Fixed` diff --git a/main.nf b/main.nf index c148c292..05a7b401 100644 --- a/main.nf +++ b/main.nf @@ -1,4 +1,4 @@ -#!/usr/bin/env nextflow +ch_bamtools_filter_config#!/usr/bin/env nextflow /* ======================================================================================== nf-core/atacseq @@ -124,103 +124,54 @@ if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)){ //////////////////////////////////////////////////// // Pipeline config -output_docs_ch = Channel.fromPath("$baseDir/docs/output.md", checkIfExists: true) +ch_output_docs = Channel.file("$baseDir/docs/output.md", checkIfExists: true) // JSON files required by BAMTools for alignment filtering if (params.singleEnd) { - bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_se_config, checkIfExists: true) + ch_bamtools_filter_config = Channel.file(params.bamtools_filter_se_config, checkIfExists: true) } else { - bamtools_filter_config_ch = Channel.fromPath(params.bamtools_filter_pe_config, checkIfExists: true) + ch_bamtools_filter_config = Channel.file(params.bamtools_filter_pe_config, checkIfExists: true) } // Header files for MultiQC -multiqc_config_ch = Channel.fromPath(params.multiqc_config, checkIfExists: true) -mlib_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) -mlib_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) -mlib_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) -mlib_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) -mlib_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) - -mrep_peak_count_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) -mrep_frip_score_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) -mrep_peak_annotation_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) -mrep_deseq2_pca_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) -mrep_deseq2_clustering_header_ch = Channel.fromPath("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) +ch_multiqc_config = file(params.multiqc_config, checkIfExists: true) +ch_mlib_peak_count_header = Channel.file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) +ch_mlib_frip_score_header = Channel.file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) +ch_mlib_peak_annotation_header = Channel.file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) +ch_mlib_deseq2_pca_header = Channel.file("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) +ch_mlib_deseq2_clustering_header = Channel.file("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) + +ch_mrep_peak_count_header = Channel.file("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) +ch_mrep_frip_score_header = Channel.file("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) +ch_mrep_peak_annotation_header = Channel.file("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) +ch_mrep_deseq2_pca_header = Channel.file("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) +ch_mrep_deseq2_clustering_header = Channel.file("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) //////////////////////////////////////////////////// /* -- VALIDATE INPUTS -- */ //////////////////////////////////////////////////// // Validate inputs -if (params.design){ - design_csv = Channel - .fromPath(params.design, checkIfExists: true) - .ifEmpty { exit 1, "Samples design file not found: ${params.design}" } -} else { - exit 1, "Samples design file not specified!" -} +if (params.design) { ch_design = file(params.design, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } +if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } +if (params.gene_bed) { ch_gene_bed = file(params.gene.bed, checkIfExists: true) } +if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } +if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } if (params.fasta){ lastPath = params.fasta.lastIndexOf(File.separator) bwa_base = params.fasta.substring(lastPath+1) - - Channel - .fromPath(params.fasta, checkIfExists: true) - .ifEmpty { exit 1, "Fasta file not found: ${params.fasta}" } - .into { fasta_bwa_index; - fasta_genome_filter; - fasta_markdup_metrics; - fasta_mlib_metrics; - fasta_mlib_macs_annotate; - fasta_mlib_macs_consensus_annotate; - fasta_mrep_macs_annotate; - fasta_mrep_macs_consensus_annotate; - fasta_igv } + ch_fasta = file(params.fasta, checkIfExists: true) } else { exit 1, "Fasta file not specified!" } -if (params.gtf){ - Channel - .fromPath(params.gtf, checkIfExists: true) - .ifEmpty { exit 1, "GTF annotation file not found: ${params.gtf}" } - .into { gtf_gene_bed; - gtf_mlib_macs_annotate; - gtf_mlib_macs_consensus_annotate; - gtf_mrep_macs_annotate; - gtf_mrep_macs_consensus_annotate } -} else { - exit 1, "GTF annotation file not specified!" -} - if (params.bwa_index){ lastPath = params.bwa_index.lastIndexOf(File.separator) bwa_dir = params.bwa_index.substring(0,lastPath+1) bwa_base = params.bwa_index.substring(lastPath+1) - - bwa_index = Channel + ch_bwa_index = Channel .fromPath(bwa_dir, checkIfExists: true) - .ifEmpty { exit 1, "BWA index directory not found: ${bwa_dir}" } -} - -if (params.gene_bed){ - gene_bed = Channel - .fromPath(params.gene_bed, checkIfExists: true) - .ifEmpty { exit 1, "Gene BED annotation file not found: ${params.gene_bed}" } -} - -if (params.tss_bed){ - tss_bed = Channel - .fromPath(params.tss_bed, checkIfExists: true) - .ifEmpty { exit 1, "TSS BED annotation file not found: ${params.tss_bed}" } -} - -if (params.blacklist) { - blacklist = Channel - .fromPath(params.blacklist, checkIfExists: true) - .ifEmpty { exit 1, "Blacklist file not found: ${params.blacklist}" } -} else { - blacklist = Channel.empty() } //////////////////////////////////////////////////// @@ -327,7 +278,7 @@ process reformat_design { tag "$design" input: - file design from design_csv + file design from ch_design output: file "*.csv" into reformat_design @@ -389,10 +340,10 @@ if (!params.bwa_index){ saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' input: - file fasta from fasta_bwa_index + file fasta from ch_fasta output: - file "BWAIndex" into bwa_index + file "BWAIndex" into ch_bwa_index script: """ @@ -411,10 +362,10 @@ if (!params.gene_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file gtf from gtf_gene_bed + file gtf from ch_gtf output: - file "*.bed" into gene_bed + file "*.bed" into ch_gene_bed script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ @@ -432,10 +383,10 @@ if (!params.tss_bed){ publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file bed from gene_bed + file bed from ch_gene_bed output: - file "*.bed" into tss_bed + file "*.bed" into ch_tss_bed script: """ @@ -452,8 +403,8 @@ process makeGenomeFilter { publishDir "${params.outdir}/reference_genome", mode: 'copy' input: - file fasta from fasta_genome_filter - file blacklist from blacklist + file fasta from ch_fasta + file blacklist from ch_blacklist output: file "$fasta" into genome_fasta // FASTA FILE FOR IGV @@ -464,9 +415,9 @@ process makeGenomeFilter { genome_sizes_mrep_bigwig script: - blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" - name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" - mito_filter = params.keepMito ? "" : name_filter + def blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" + def name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" + def mito_filter = params.keepMito ? "" : name_filter """ samtools faidx $fasta get_autosomes.py ${fasta}.fai ${fasta}.autosomes.txt @@ -588,7 +539,7 @@ process bwa_mem { input: set val(name), file(reads) from trimmed_reads - file index from bwa_index.first() + file index from ch_bwa_index.collect() output: set val(name), file("*.bam") into bwa_bam @@ -743,7 +694,7 @@ process merge_library_filter { input: set val(name), file(bam) from mlib_bam_filter file bed from genome_filter_regions.collect() - file bamtools_filter_config from bamtools_filter_config_ch.collect() + file bamtools_filter_config from ch_bamtools_filter_config output: set val(name), file("*.{bam,bam.bai}") into mlib_filter_bam @@ -856,7 +807,7 @@ process merge_library_collectmetrics { input: set val(name), file(bam) from mlib_rm_orphan_bam_metrics - file fasta from fasta_mlib_metrics.collect() + file fasta from ch_fasta output: file "*_metrics" into mlib_collectmetrics_mqc @@ -928,8 +879,8 @@ process merge_library_macs { input: set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_macs.join(mlib_rm_orphan_flagstat_macs, by: [0]) - file mlib_peak_count_header from mlib_peak_count_header_ch.collect() - file mlib_frip_score_header from mlib_frip_score_header_ch.collect() + file mlib_peak_count_header from ch_mlib_peak_count_header + file mlib_frip_score_header from ch_mlib_frip_score_header output: set val(name), file("*.{bed,xls,gappedPeak}") into mlib_macs_output @@ -974,8 +925,8 @@ process merge_library_macs_annotate { input: set val(name), file(peak) from mlib_macs_peaks_homer - file fasta from fasta_mlib_macs_annotate.collect() - file gtf from gtf_mlib_macs_annotate.collect() + file fasta from ch_fasta + file gtf from ch_gtf output: file "*.txt" into mlib_macs_annotate @@ -1003,7 +954,7 @@ process merge_library_macs_qc { input: file peaks from mlib_macs_peaks_qc.collect{ it[1] } file annos from mlib_macs_annotate.collect() - file mlib_peak_annotation_header from mlib_peak_annotation_header_ch.collect() + file mlib_peak_annotation_header from ch_mlib_peak_annotation_header output: file "*.{txt,pdf}" into mlib_macs_qc @@ -1084,8 +1035,8 @@ process merge_library_macs_consensus_annotate { input: file bed from mlib_macs_consensus_bed file bool from mlib_macs_consensus_bool - file fasta from fasta_mlib_macs_consensus_annotate - file gtf from gtf_mlib_macs_consensus_annotate + file fasta from ch_fasta + file gtf from ch_gtf output: file "*.annotatePeaks.txt" into mlib_macs_consensus_annotate @@ -1116,8 +1067,8 @@ process merge_library_macs_consensus_deseq { input: file bams from mlib_name_bam_mlib_counts.collect{ it[1] } file saf from mlib_macs_consensus_saf.collect() - file mlib_deseq2_pca_header from mlib_deseq2_pca_header_ch.collect() - file mlib_deseq2_clustering_header from mlib_deseq2_clustering_header_ch.collect() + file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header + file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header output: file "*featureCounts.txt" into mlib_macs_consensus_counts @@ -1162,7 +1113,7 @@ process merge_library_ataqv { input: set val(name), file(bam), file(peak) from mlib_bam_ataqv.join(mlib_macs_peaks_ataqv, by: [0]) - file tss_bed from tss_bed.collect() + file tss_bed from ch_tss_bed file autosomes from genome_autosomes.collect() output: @@ -1352,8 +1303,8 @@ process merge_replicate_macs { input: set val(name), file(bam), file(flagstat) from mrep_bam_macs.join(mrep_flagstat_macs, by: [0]) - file mrep_peak_count_header from mrep_peak_count_header_ch.collect() - file mrep_frip_score_header from mrep_frip_score_header_ch.collect() + file mrep_peak_count_header from ch_mrep_peak_count_header + file mrep_frip_score_header from ch_mrep_frip_score_header output: file "*.{bed,xls,gappedPeak}" into mrep_macs_output @@ -1397,8 +1348,8 @@ process merge_replicate_macs_annotate { input: set val(name), file(peak) from mrep_macs_peaks_homer - file fasta from fasta_mrep_macs_annotate.collect() - file gtf from gtf_mrep_macs_annotate.collect() + file fasta from ch_fasta + file gtf from ch_gtf output: file "*.txt" into mrep_macs_annotate @@ -1426,7 +1377,7 @@ process merge_replicate_macs_qc { input: file peaks from mrep_macs_peaks_qc.collect{ it[1] } file annos from mrep_macs_annotate.collect() - file mrep_peak_annotation_header from mrep_peak_annotation_header_ch.collect() + file mrep_peak_annotation_header from ch_mrep_peak_annotation_header output: file "*.{txt,pdf}" into mrep_macs_qc @@ -1507,8 +1458,8 @@ process merge_replicate_macs_consensus_annotate { input: file bed from mrep_macs_consensus_bed file bool from mrep_macs_consensus_bool - file fasta from fasta_mrep_macs_consensus_annotate - file gtf from gtf_mrep_macs_consensus_annotate + file fasta from ch_fasta + file gtf from ch_gtf output: file "*.annotatePeaks.txt" into mrep_macs_consensus_annotate @@ -1539,8 +1490,8 @@ process merge_replicate_macs_consensus_deseq { input: file bams from mlib_name_bam_mrep_counts.collect{ it[1] } file saf from mrep_macs_consensus_saf.collect() - file mrep_deseq2_pca_header from mrep_deseq2_pca_header_ch.collect() - file mrep_deseq2_clustering_header from mrep_deseq2_clustering_header_ch.collect() + file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header + file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header output: file "*featureCounts.txt" into mrep_macs_consensus_counts @@ -1590,7 +1541,7 @@ process igv { publishDir "${params.outdir}/igv", mode: 'copy' input: - file fasta from fasta_igv.collect() + file fasta from ch_fasta file ('bwa/mergedLibrary/bigwig/*') from mlib_bigwig_igv.collect() file ('bwa/mergedLibrary/macs/*') from mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) @@ -1683,7 +1634,7 @@ process multiqc { publishDir "${params.outdir}/multiqc", mode: 'copy' input: - file multiqc_config from multiqc_config_ch.collect() + file multiqc_config from ch_multiqc_config file ('fastqc/*') from fastqc_reports_mqc.collect() file ('trimgalore/*') from trimgalore_results_mqc.collect() @@ -1741,7 +1692,7 @@ process output_documentation { publishDir "${params.outdir}/Documentation", mode: 'copy' input: - file output_docs from output_docs_ch + file output_docs from ch_output_docs output: file "results_description.html" From 9972c0483c2c5b927524d8cd0f3bea201525af77 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:07:17 +0100 Subject: [PATCH 024/149] Update resource requirements --- conf/base.config | 10 +++++++--- 1 file changed, 7 insertions(+), 3 deletions(-) diff --git a/conf/base.config b/conf/base.config index 4ac857c3..9d01790c 100755 --- a/conf/base.config +++ b/conf/base.config @@ -12,7 +12,7 @@ process { cpus = { check_max( 1 * task.attempt, 'cpus' ) } - memory = { check_max( 2.GB * task.attempt, 'memory' ) } + memory = { check_max( 8.GB * task.attempt, 'memory' ) } time = { check_max( 2.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } @@ -20,12 +20,17 @@ process { maxErrors = '-1' // Process-specific resource requirements + withLabel:process_low { + cpus = { check_max( 2 * task.attempt, 'cpus' ) } + memory = { check_max( 16.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } + } withLabel:process_medium { cpus = { check_max( 6 * task.attempt, 'cpus' ) } memory = { check_max( 48.GB * task.attempt, 'memory' ) } time = { check_max( 8.h * task.attempt, 'time' ) } } - withLabel:process_big { + withLabel:process_high { cpus = { check_max( 12 * task.attempt, 'cpus' ) } memory = { check_max( 96.GB * task.attempt, 'memory' ) } time = { check_max( 16.h * task.attempt, 'time' ) } @@ -36,7 +41,6 @@ process { withName:get_software_versions { cache = false } - } // Defaults config params, may be overwritten by later configs From f0ede9eb976233f983937aea4a78291a51ea4e50 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:07:32 +0100 Subject: [PATCH 025/149] Update process names --- main.nf | 37 +++++++++++++++++++------------------ 1 file changed, 19 insertions(+), 18 deletions(-) diff --git a/main.nf b/main.nf index 05a7b401..ac5da048 100644 --- a/main.nf +++ b/main.nf @@ -274,18 +274,19 @@ if (!params.macs_gsize){ /* * PREPROCESSING - REFORMAT DESIGN FILE AND CHECK VALIDITY */ -process reformat_design { +process CheckDesign { tag "$design" + publishDir "${params.outdir}/pipeline_info", mode: 'copy' input: file design from ch_design output: - file "*.csv" into reformat_design + file "*.csv" into ch_design_reads_csv script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ """ - reformat_design.py $design design_reformat.csv + check_design.py $design design_reads.csv """ } @@ -293,19 +294,19 @@ process reformat_design { * Create channels for input fastq files */ if (params.singleEnd) { - reformat_design.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + ch_design_reads_csv.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } + .into { design_replicates_exist; + design_multiple_samples; + raw_reads_fastqc; + raw_reads_trimgalore } } else { - reformat_design.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + ch_design_reads_csv.splitCsv(header:true, sep:',') + .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } + .into { design_replicates_exist; + design_multiple_samples; + raw_reads_fastqc; + raw_reads_trimgalore } } // Boolean value for replicates existing in design @@ -333,9 +334,9 @@ multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].j * PREPROCESSING - Build BWA index */ if (!params.bwa_index){ - process makeBWAindex { + process MakeBWAIndex { tag "$fasta" - label 'process_big' + label 'process_high' publishDir path: { params.saveGenomeIndex ? "${params.outdir}/reference_genome" : params.outdir }, saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' @@ -535,7 +536,7 @@ if (params.skipTrimming){ */ process bwa_mem { tag "$name" - label 'process_big' + label 'process_high' input: set val(name), file(reads) from trimmed_reads From 1ef0dca0fcf6798feb5f54552618e62ba56a9470 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:08:18 +0100 Subject: [PATCH 026/149] Rename check_design --- bin/{reformat_design.py => check_design.py} | 0 1 file changed, 0 insertions(+), 0 deletions(-) rename bin/{reformat_design.py => check_design.py} (100%) diff --git a/bin/reformat_design.py b/bin/check_design.py similarity index 100% rename from bin/reformat_design.py rename to bin/check_design.py From 99ffe090de76455d9ca30ce459ab74ecef9d0023 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:24:30 +0100 Subject: [PATCH 027/149] Update Zenodo ID --- README.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/README.md b/README.md index 8b5437c0..9de52ed2 100644 --- a/README.md +++ b/README.md @@ -6,7 +6,7 @@ [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) -[![DOI](https://zenodo.org/badge/156259074.svg)](https://zenodo.org/badge/latestdoi/156259074) +[![DOI](https://zenodo.org/badge/DOI/10.5281/zenodo.2634133.svg)](https://doi.org/10.5281/zenodo.2634133) ## Introduction From b614910482f7d3a9545fe20b8454ab474baa66e3 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:27:42 +0100 Subject: [PATCH 028/149] Fix copy bug --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index ac5da048..51341010 100644 --- a/main.nf +++ b/main.nf @@ -1,4 +1,4 @@ -ch_bamtools_filter_config#!/usr/bin/env nextflow +#!/usr/bin/env nextflow /* ======================================================================================== nf-core/atacseq From 14f0672356911fc5f47d6b3d16f19a34f7d99b37 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 22:45:42 +0100 Subject: [PATCH 029/149] Add def everywhere --- main.nf | 198 ++++++++++++++++++++++++++++---------------------------- 1 file changed, 99 insertions(+), 99 deletions(-) diff --git a/main.nf b/main.nf index 51341010..8ca5a0ef 100644 --- a/main.nf +++ b/main.nf @@ -358,7 +358,7 @@ if (!params.bwa_index){ * PREPROCESSING - Generate gene BED file */ if (!params.gene_bed){ - process makeGeneBED { + process MakeGeneBED { tag "$gtf" publishDir "${params.outdir}/reference_genome", mode: 'copy' @@ -379,7 +379,7 @@ if (!params.gene_bed){ * PREPROCESSING - Generate TSS BED file */ if (!params.tss_bed){ - process makeTSSBED { + process MakeTSSBED { tag "$bed" publishDir "${params.outdir}/reference_genome", mode: 'copy' @@ -399,7 +399,7 @@ if (!params.tss_bed){ /* * PREPROCESSING - Prepare genome intervals for filtering */ -process makeGenomeFilter { +process MakeGenomeFilter { tag "$fasta" publishDir "${params.outdir}/reference_genome", mode: 'copy' @@ -438,7 +438,7 @@ process makeGenomeFilter { /* * STEP 1 - FastQC */ -process fastqc { +process FastQC { tag "$name" label 'process_medium' publishDir "${params.outdir}/fastqc", mode: 'copy', @@ -483,7 +483,7 @@ if (params.skipTrimming){ trimgalore_results_mqc = [] trimgalore_fastqc_reports_mqc = [] } else { - process trim_galore { + process TrimGalore { tag "$name" label 'process_long' publishDir "${params.outdir}/trim_galore", mode: 'copy', @@ -504,10 +504,10 @@ if (params.skipTrimming){ script: // Added soft-links to original fastqs for consistent naming in MultiQC - c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' - c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' - tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' - tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' + def c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' + def c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' + def tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' + def tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' if (params.singleEnd) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz @@ -534,7 +534,7 @@ if (params.skipTrimming){ /* * STEP 3.1 - Align read 1 with bwa */ -process bwa_mem { +process BWAmem { tag "$name" label 'process_high' @@ -546,8 +546,8 @@ process bwa_mem { set val(name), file("*.bam") into bwa_bam script: - prefix="${name}.Lb" - rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + def prefix="${name}.Lb" + def rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" """ bwa mem -t $task.cpus -M -R $rg ${index}/${bwa_base} $reads | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - """ @@ -556,7 +556,7 @@ process bwa_mem { /* * STEP 3.2 - Convert .bam to coordinate sorted .bam */ -process sort_bam { +process SortBAM { tag "$name" label 'process_medium' if (params.saveAlignedIntermediates) { @@ -576,7 +576,7 @@ process sort_bam { file "*.{flagstat,idxstats,stats}" into sort_bam_flagstat_mqc script: - prefix="${name}.Lb" + def prefix="${name}.Lb" """ samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam samtools index ${prefix}.sorted.bam @@ -602,7 +602,7 @@ sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } .map { it -> [ it[0], it[1].flatten() ] } .set { sort_bam_mlib } -process merge_library { +process MergeLibrary { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -624,11 +624,11 @@ process merge_library { file "*.txt" into mlib_metrics_mqc script: - prefix="${name}.mLb.mkD" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def prefix="${name}.mLb.mkD" + def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 if (!task.memory){ log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - avail_mem = 3 } else { avail_mem = task.memory.toGiga() } @@ -678,7 +678,7 @@ process merge_library { /* * STEP 4.2 Filter BAM file at merged library-level */ -process merge_library_filter { +process MergeLibraryFilter { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -703,11 +703,11 @@ process merge_library_filter { file "*.{idxstats,stats}" into mlib_filter_stats_mqc script: - prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" - filter_params = params.singleEnd ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" - dup_params = params.keepDups ? "" : "-F 0x0400" - multimap_params = params.keepMultiMap ? "" : "-q 1" - name_sort_bam = params.singleEnd ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" + def prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" + def filter_params = params.singleEnd ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" + def dup_params = params.keepDups ? "" : "-F 0x0400" + def multimap_params = params.keepMultiMap ? "" : "-q 1" + def name_sort_bam = params.singleEnd ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" """ samtools view \\ $filter_params \\ @@ -743,7 +743,7 @@ if (params.singleEnd){ mlib_rm_orphan_flagstat_mqc } mlib_filter_stats_mqc.set { mlib_rm_orphan_stats_mqc } } else { - process merge_library_rm_orphan { + process MergeLibraryRemoveOrphanReads { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -772,7 +772,7 @@ if (params.singleEnd){ file "*.{idxstats,stats}" into mlib_rm_orphan_stats_mqc script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - prefix="${name}.mLb.clN" + def prefix="${name}.mLb.clN" """ bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs @@ -796,7 +796,7 @@ if (params.singleEnd){ /* * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries */ -process merge_library_collectmetrics { +process MergeLibraryCollectMultipleMetrics { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -815,10 +815,10 @@ process merge_library_collectmetrics { file "*.pdf" into mlib_collectmetrics_pdf script: - prefix="${name}.mLb.clN" + def prefix="${name}.mLb.clN" + def avail_mem = 3 if (!task.memory){ - log.info "[Picard CollectMultipleMetrics] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - avail_mem = 3 + log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." } else { avail_mem = task.memory.toGiga() } @@ -835,7 +835,7 @@ process merge_library_collectmetrics { /* * STEP 4.4.2 Read depth normalised bigWig */ -process merge_library_bigwig { +process MergeLibraryBigWig { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', @@ -854,9 +854,9 @@ process merge_library_bigwig { file "*.txt" into mlib_bigwig_scale script: - prefix="${name}.mLb.clN" - pe_fragment = params.singleEnd ? "" : "-pc" - extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + def prefix="${name}.mLb.clN" + def pe_fragment = params.singleEnd ? "" : "-pc" + def extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt @@ -869,7 +869,7 @@ process merge_library_bigwig { /* * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score */ -process merge_library_macs { +process MergeLibraryMACSCallPeak { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy', @@ -895,10 +895,10 @@ process merge_library_macs { when: params.macs_gsize script: - prefix="${name}.mLb.clN" - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - broad = params.narrowPeak ? '' : "--broad" - format = params.singleEnd ? "BAM" : "BAMPE" + def prefix="${name}.mLb.clN" + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def broad = params.narrowPeak ? '' : "--broad" + def format = params.singleEnd ? "BAM" : "BAMPE" """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -919,7 +919,7 @@ process merge_library_macs { /* * STEP 4.5.2 Annotate peaks with HOMER */ -process merge_library_macs_annotate { +process MergeLibraryAnnotatePeaks { tag "$name" label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy' @@ -935,7 +935,7 @@ process merge_library_macs_annotate { when: params.macs_gsize script: - prefix="${name}.mLb.clN" + def prefix="${name}.mLb.clN" """ annotatePeaks.pl $peak \\ $fasta \\ @@ -948,7 +948,7 @@ process merge_library_macs_annotate { /* * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process merge_library_macs_qc { +process MergeLibraryPeakQC { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/qc", mode: 'copy' @@ -964,8 +964,8 @@ process merge_library_macs_qc { when: params.macs_gsize script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix='mLb.clN' - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def suffix='mLb.clN' + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" """ plot_macs_qc.r -i ${peaks.join(',')} \\ -s ${peaks.join(',').replaceAll(".${suffix}_peaks${peakext}","")} \\ @@ -984,7 +984,7 @@ process merge_library_macs_qc { /* * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process merge_library_macs_consensus { +process MergeLibraryCreateConsensusPeakSet { publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' input: @@ -1000,12 +1000,12 @@ process merge_library_macs_consensus { when: params.macs_gsize && (multiple_samples || replicates_exist) script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix='mLb.clN' - prefix="consensus_peaks.${suffix}" - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrowPeak ? "--is_narrow_peak" : "" + def suffix='mLb.clN' + def prefix="consensus_peaks.${suffix}" + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') + def collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + def expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt @@ -1029,7 +1029,7 @@ process merge_library_macs_consensus { /* * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process merge_library_macs_consensus_annotate { +process MergeLibraryAnnotateConsensusPeakSet { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' @@ -1045,7 +1045,7 @@ process merge_library_macs_consensus_annotate { when: params.macs_gsize && (multiple_samples || replicates_exist) script: - prefix="consensus_peaks.mLb.clN" + def prefix="consensus_peaks.mLb.clN" """ annotatePeaks.pl $bed \\ $fasta \\ @@ -1061,7 +1061,7 @@ process merge_library_macs_consensus_annotate { /* * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process merge_library_macs_consensus_deseq { +process MergeLibraryDeseqConsensusPeakSet { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus/deseq2", mode: 'copy' @@ -1083,10 +1083,10 @@ process merge_library_macs_consensus_deseq { when: params.macs_gsize && multiple_samples && replicates_exist script: - prefix="consensus_peaks.mLb.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - pe_params = params.singleEnd ? '' : "-p --donotsort" + def prefix="consensus_peaks.mLb.clN" + def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + def pe_params = params.singleEnd ? '' : "-p --donotsort" """ featureCounts -F SAF \\ -O \\ @@ -1107,7 +1107,7 @@ process merge_library_macs_consensus_deseq { /* * STEP 4.6.1 Run ataqv on BAM file and corresponding peaks */ -process merge_library_ataqv { +process MergeLibraryAtaqv { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' @@ -1121,8 +1121,8 @@ process merge_library_ataqv { file "*.json" into mlib_ataqv script: - peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" - mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" + def peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" + def mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" """ ataqv --threads $task.cpus \\ $peak_param \\ @@ -1140,7 +1140,7 @@ process merge_library_ataqv { /* * STEP 4.6.2 run ataqv mkarv on all JSON files to render web app */ -process merge_library_ataqv_mkarv { +process MergeLibraryAtaqvMkarv { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' @@ -1175,7 +1175,7 @@ mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] .map { it -> [ it[0], it[1].flatten() ] } .set { mlib_rm_orphan_bam_mrep } -process merge_replicate { +process MergeReplicate { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', @@ -1202,11 +1202,11 @@ process merge_replicate { when: !params.skipMergeReplicates && replicates_exist script: - prefix="${name}.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def prefix="${name}.mRp.clN" + def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def avail_mem = 3 if (!task.memory){ log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." - avail_mem = 3 } else { avail_mem = task.memory.toGiga() } @@ -1257,7 +1257,7 @@ process merge_replicate { /* * STEP 5.2.1 Read depth normalised bigWig */ -process merge_replicate_bigwig { +process MergeReplicateBigWig { tag "$name" label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', @@ -1278,9 +1278,9 @@ process merge_replicate_bigwig { when: !params.skipMergeReplicates && replicates_exist script: - prefix="${name}.mRp.clN" - pe_fragment = params.singleEnd ? "" : "-pc" - extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + def prefix="${name}.mRp.clN" + def pe_fragment = params.singleEnd ? "" : "-pc" + def extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt @@ -1293,7 +1293,7 @@ process merge_replicate_bigwig { /* * STEP 5.2.2 Call peaks with MACS2 and calculate FRiP score */ -process merge_replicate_macs { +process MergeReplicateMACSCallPeak { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy', @@ -1318,10 +1318,10 @@ process merge_replicate_macs { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: - prefix="${name}.mRp.clN" - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - broad = params.narrowPeak ? '' : "--broad" - format = params.singleEnd ? "BAM" : "BAMPE" + def prefix="${name}.mRp.clN" + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def broad = params.narrowPeak ? '' : "--broad" + def format = params.singleEnd ? "BAM" : "BAMPE" """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -1342,7 +1342,7 @@ process merge_replicate_macs { /* * STEP 5.2.3 Annotate peaks with HOMER */ -process merge_replicate_macs_annotate { +process MergeReplicateAnnotatePeaks { tag "$name" label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy' @@ -1358,7 +1358,7 @@ process merge_replicate_macs_annotate { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: - prefix="${name}.mRp.clN" + def prefix="${name}.mRp.clN" """ annotatePeaks.pl $peak \\ $fasta \\ @@ -1371,7 +1371,7 @@ process merge_replicate_macs_annotate { /* * STEP 5.2.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ -process merge_replicate_macs_qc { +process MergeReplicatePeakQC { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/qc", mode: 'copy' @@ -1387,8 +1387,8 @@ process merge_replicate_macs_qc { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - suffix='mRp.clN' - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def suffix='mRp.clN' + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" """ plot_macs_qc.r -i ${peaks.join(',')} \\ -s ${peaks.join(',').replaceAll(".${suffix}_peaks${peakext}","")} \\ @@ -1407,7 +1407,7 @@ process merge_replicate_macs_qc { /* * STEP 5.2.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process merge_replicate_macs_consensus { +process MergeReplicateCreateConsensusPeakSet { publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' input: @@ -1423,12 +1423,12 @@ process merge_replicate_macs_consensus { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - suffix='mRp.clN' - prefix="consensus_peaks.${suffix}" - peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') - collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - expandparam = params.narrowPeak ? "--is_narrow_peak" : "" + def suffix='mRp.clN' + def prefix="consensus_peaks.${suffix}" + def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" + def mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') + def collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + def expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt @@ -1452,7 +1452,7 @@ process merge_replicate_macs_consensus { /* * STEP 5.2.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process merge_replicate_macs_consensus_annotate { +process MergeReplicateAnnotateConsensusPeakSet { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' @@ -1468,7 +1468,7 @@ process merge_replicate_macs_consensus_annotate { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: - prefix="consensus_peaks.mRp.clN" + def prefix="consensus_peaks.mRp.clN" """ annotatePeaks.pl $bed \\ $fasta \\ @@ -1484,7 +1484,7 @@ process merge_replicate_macs_consensus_annotate { /* * STEP 5.2.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process merge_replicate_macs_consensus_deseq { +process MergeReplicateDeseqConsensusPeakSet { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus/deseq2", mode: 'copy' @@ -1506,10 +1506,10 @@ process merge_replicate_macs_consensus_deseq { when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: - prefix="consensus_peaks.mRp.clN" - bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - pe_params = params.singleEnd ? '' : "-p --donotsort" + def prefix="consensus_peaks.mRp.clN" + def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + def bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + def pe_params = params.singleEnd ? '' : "-p --donotsort" """ featureCounts -F SAF \\ -O \\ @@ -1538,7 +1538,7 @@ process merge_replicate_macs_consensus_deseq { /* * STEP 6 - Create IGV session file */ -process igv { +process IGV { publishDir "${params.outdir}/igv", mode: 'copy' input: @@ -1558,7 +1558,7 @@ process igv { file "*.{txt,xml}" into igv_session script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - outdir_abspath = new File(params.outdir).getCanonicalPath().toString() + def outdir_abspath = new File(params.outdir).getCanonicalPath().toString() """ igv_get_files.sh ./ mLb $outdir_abspath > igv_files.txt igv_get_files.sh ./ mRp $outdir_abspath >> igv_files.txt @@ -1631,7 +1631,7 @@ ${summary.collect { k,v -> "
$k
${v ?: ' Date: Mon, 30 Sep 2019 22:49:29 +0100 Subject: [PATCH 030/149] Replace Channel.file with file --- main.nf | 28 ++++++++++++++-------------- 1 file changed, 14 insertions(+), 14 deletions(-) diff --git a/main.nf b/main.nf index 8ca5a0ef..f317b297 100644 --- a/main.nf +++ b/main.nf @@ -124,28 +124,28 @@ if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)){ //////////////////////////////////////////////////// // Pipeline config -ch_output_docs = Channel.file("$baseDir/docs/output.md", checkIfExists: true) +ch_output_docs = file("$baseDir/docs/output.md", checkIfExists: true) // JSON files required by BAMTools for alignment filtering if (params.singleEnd) { - ch_bamtools_filter_config = Channel.file(params.bamtools_filter_se_config, checkIfExists: true) + ch_bamtools_filter_config = file(params.bamtools_filter_se_config, checkIfExists: true) } else { - ch_bamtools_filter_config = Channel.file(params.bamtools_filter_pe_config, checkIfExists: true) + ch_bamtools_filter_config = file(params.bamtools_filter_pe_config, checkIfExists: true) } // Header files for MultiQC ch_multiqc_config = file(params.multiqc_config, checkIfExists: true) -ch_mlib_peak_count_header = Channel.file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) -ch_mlib_frip_score_header = Channel.file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) -ch_mlib_peak_annotation_header = Channel.file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) -ch_mlib_deseq2_pca_header = Channel.file("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) -ch_mlib_deseq2_clustering_header = Channel.file("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) - -ch_mrep_peak_count_header = Channel.file("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) -ch_mrep_frip_score_header = Channel.file("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) -ch_mrep_peak_annotation_header = Channel.file("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) -ch_mrep_deseq2_pca_header = Channel.file("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) -ch_mrep_deseq2_clustering_header = Channel.file("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) +ch_mlib_peak_count_header = file("$baseDir/assets/multiqc/mlib_peak_count_header.txt", checkIfExists: true) +ch_mlib_frip_score_header = file("$baseDir/assets/multiqc/mlib_frip_score_header.txt", checkIfExists: true) +ch_mlib_peak_annotation_header = file("$baseDir/assets/multiqc/mlib_peak_annotation_header.txt", checkIfExists: true) +ch_mlib_deseq2_pca_header = file("$baseDir/assets/multiqc/mlib_deseq2_pca_header.txt", checkIfExists: true) +ch_mlib_deseq2_clustering_header = file("$baseDir/assets/multiqc/mlib_deseq2_clustering_header.txt", checkIfExists: true) + +ch_mrep_peak_count_header = file("$baseDir/assets/multiqc/mrep_peak_count_header.txt", checkIfExists: true) +ch_mrep_frip_score_header = file("$baseDir/assets/multiqc/mrep_frip_score_header.txt", checkIfExists: true) +ch_mrep_peak_annotation_header = file("$baseDir/assets/multiqc/mrep_peak_annotation_header.txt", checkIfExists: true) +ch_mrep_deseq2_pca_header = file("$baseDir/assets/multiqc/mrep_deseq2_pca_header.txt", checkIfExists: true) +ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clustering_header.txt", checkIfExists: true) //////////////////////////////////////////////////// /* -- VALIDATE INPUTS -- */ From a85233f0c0b5dd101d0e8e64659326db6d550146 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 30 Sep 2019 23:04:17 +0100 Subject: [PATCH 031/149] Update names --- main.nf | 45 ++++++++++++++++++++++++++++++--------------- 1 file changed, 30 insertions(+), 15 deletions(-) diff --git a/main.nf b/main.nf index f317b297..0904ad61 100644 --- a/main.nf +++ b/main.nf @@ -334,7 +334,7 @@ multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].j * PREPROCESSING - Build BWA index */ if (!params.bwa_index){ - process MakeBWAIndex { + process BWAIndex { tag "$fasta" label 'process_high' publishDir path: { params.saveGenomeIndex ? "${params.outdir}/reference_genome" : params.outdir }, @@ -360,6 +360,7 @@ if (!params.bwa_index){ if (!params.gene_bed){ process MakeGeneBED { tag "$gtf" + label 'process_low' publishDir "${params.outdir}/reference_genome", mode: 'copy' input: @@ -892,7 +893,8 @@ process MergeLibraryMACSCallPeak { mlib_macs_peaks_igv file "*_mqc.tsv" into mlib_macs_peaks_mqc - when: params.macs_gsize + when: + params.macs_gsize script: def prefix="${name}.mLb.clN" @@ -932,7 +934,8 @@ process MergeLibraryAnnotatePeaks { output: file "*.txt" into mlib_macs_annotate - when: params.macs_gsize + when: + params.macs_gsize script: def prefix="${name}.mLb.clN" @@ -961,7 +964,8 @@ process MergeLibraryPeakQC { file "*.{txt,pdf}" into mlib_macs_qc file "*.tsv" into mlib_macs_qc_mqc - when: params.macs_gsize + when: + params.macs_gsize script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ def suffix='mLb.clN' @@ -997,7 +1001,8 @@ process MergeLibraryCreateConsensusPeakSet { file "*.saf" into mlib_macs_consensus_saf file "*.intersect.{txt,plot.pdf}" into mlib_macs_consensus_intersect - when: params.macs_gsize && (multiple_samples || replicates_exist) + when: + params.macs_gsize && (multiple_samples || replicates_exist) script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ def suffix='mLb.clN' @@ -1042,7 +1047,8 @@ process MergeLibraryAnnotateConsensusPeakSet { output: file "*.annotatePeaks.txt" into mlib_macs_consensus_annotate - when: params.macs_gsize && (multiple_samples || replicates_exist) + when: + params.macs_gsize && (multiple_samples || replicates_exist) script: def prefix="consensus_peaks.mLb.clN" @@ -1080,7 +1086,8 @@ process MergeLibraryDeseqConsensusPeakSet { file "*vs*/*.bed" into mlib_macs_consensus_deseq_comp_bed_igv file "*.tsv" into mlib_macs_consensus_deseq_mqc - when: params.macs_gsize && multiple_samples && replicates_exist + when: + params.macs_gsize && multiple_samples && replicates_exist script: def prefix="consensus_peaks.mLb.clN" @@ -1199,7 +1206,8 @@ process MergeReplicate { file "*.{idxstats,stats}" into mrep_stats_mqc file "*.txt" into mrep_metrics_mqc - when: !params.skipMergeReplicates && replicates_exist + when: + !params.skipMergeReplicates && replicates_exist script: def prefix="${name}.mRp.clN" @@ -1275,7 +1283,8 @@ process MergeReplicateBigWig { file "*.bigWig" into mrep_bigwig_igv file "*.txt" into mrep_bigwig_scale - when: !params.skipMergeReplicates && replicates_exist + when: + !params.skipMergeReplicates && replicates_exist script: def prefix="${name}.mRp.clN" @@ -1315,7 +1324,8 @@ process MergeReplicateMACSCallPeak { mrep_macs_peaks_igv file "*_mqc.tsv" into mrep_macs_peak_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: def prefix="${name}.mRp.clN" @@ -1355,7 +1365,8 @@ process MergeReplicateAnnotatePeaks { output: file "*.txt" into mrep_macs_annotate - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: def prefix="${name}.mRp.clN" @@ -1384,7 +1395,8 @@ process MergeReplicatePeakQC { file "*.{txt,pdf}" into mrep_macs_qc file "*.tsv" into mrep_macs_qc_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ def suffix='mRp.clN' @@ -1420,7 +1432,8 @@ process MergeReplicateCreateConsensusPeakSet { file "*.saf" into mrep_macs_consensus_saf file "*.intersect.{txt,plot.pdf}" into mrep_macs_consensus_intersect - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ def suffix='mRp.clN' @@ -1465,7 +1478,8 @@ process MergeReplicateAnnotateConsensusPeakSet { output: file "*.annotatePeaks.txt" into mrep_macs_consensus_annotate - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: def prefix="consensus_peaks.mRp.clN" @@ -1503,7 +1517,8 @@ process MergeReplicateDeseqConsensusPeakSet { file "*vs*/*.bed" into mrep_macs_consensus_deseq_comp_bed_igv file "*.tsv" into mrep_macs_consensus_deseq_mqc - when: !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + when: + !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples script: def prefix="consensus_peaks.mRp.clN" From 25347f620696bc5147ee28fd86a9c36905177fba Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 11:29:58 +0100 Subject: [PATCH 032/149] Remove unrequired files --- assets/multiqc_config.yaml | 9 --------- bin/scrape_software_versions.pyc | Bin 1540 -> 0 bytes 2 files changed, 9 deletions(-) delete mode 100644 assets/multiqc_config.yaml delete mode 100755 bin/scrape_software_versions.pyc diff --git a/assets/multiqc_config.yaml b/assets/multiqc_config.yaml deleted file mode 100644 index 8e6246d7..00000000 --- a/assets/multiqc_config.yaml +++ /dev/null @@ -1,9 +0,0 @@ -report_comment: > - This report has been generated by the nf-core/atacseq - analysis pipeline. For information about how to interpret these results, please see the - documentation. -report_section_order: - nf-core/atacseq-software-versions: - order: -1000 - -export_plots: true diff --git a/bin/scrape_software_versions.pyc b/bin/scrape_software_versions.pyc deleted file mode 100755 index 566a29d20d3348c3ac3792a3cefc9e3082c2cc2c..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 1540 zcmZ`&UvC>l5TEtsk8_FBq=BOFfKK7Tj#}SQ9*PLA1!~0`L2V?8Ows9bJGM93yS?o0 zxlV(5$p_#Q@W!{{V&(${{B-A&1%#CYhYkf2Eb`E zkN^dP5d-FM3uFt507k(Q2O!sAv<6}gIPGzXe=}i%sYD{AUTTTQF+FXbUg|q?7jG@Rq@E zr!d+E+=h!CAHM^*1{b^DVjm8%1GnTyN8SS+GJYQ}_8fn)4}2fUpEhNKltT&diU;uG z$E!c!72XGa;FN6snKM|War90aWb%Oz*LJ7TG0yzzA@j~3dPMQgdJm}B;%{0#DmK_% zB9#`UO~-YaTcOIOe4FIoYmOTEbCKIq;|ynMB`TDn#MsWQStn8N_1VK?^AT}3H&L$C zrNF$l7Ij=^1xAZS<5=R{j-`6z9Ib2kX(KQ@KJYj*_A2*bJ0$p);EQL`%!judcM{{2 zv$UwC6^*&OP=g;uGt`Fe5H0;SN%fsGeeuk6i659MDS%bH zd^1R#!BCJX2v;5x5SiU9%z#iR_({}#-!)^p80y|}tg(_=j=gTweH3;3-D4_1->9*D zlWFWX$(e3w(3d)OHTs{JYrW$+)i@^4skN0kNs@`M)A~HlRgt_eK?>Fl zE2(U1uPb-r)E2TEax}RXmFqgmC`Ia^l$0CtD6^5S%g74q;#jL9vQvy!SBX?@tJ=mC zBD2{^^h+4g8gLoqGBf7s5tsd`(&DWuZ6=R~q4|Ug>1^B@a67zN3=;YcIA5}SXto*? zQpjQNfRGJ|q%@hf)|jR*34wK!*`!H%o0oo!TO`zIUBfj_aE_+o4Yl7`H>tv6VXU*h z^mJy|h*uSs9?m_~m~lUzt6urpGnnao>Z25yCC&}xSZM-YIMJ%EJm;TMkF@YFF@F+F z@~o(mtg1{>2$LtqR>^sGPADtuOeRLvI!BX~<9@DaR;*Ie^dHBM6Hywfp-iMWPpWG> zRpsaL7m1x7XTY;qnU(*#X;O57-7f N4ubY3J7gcTe*pU!kgfm# From 62b86a37f38efebffd11aaa1d98b13d9bfd5fb37 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 11:30:10 +0100 Subject: [PATCH 033/149] Add Preseq --- environment.yml | 1 + 1 file changed, 1 insertion(+) diff --git a/environment.yml b/environment.yml index e18a23a4..dbffed49 100755 --- a/environment.yml +++ b/environment.yml @@ -26,6 +26,7 @@ dependencies: - bamtools=2.5.1 - pysam=0.15.2 - bedtools=2.27.1 + - preseq=2.0.3 - ucsc-bedgraphtobigwig=377 - macs2=2.1.2 - homer=4.9.1 From 1dc2d50e43a6e43ca224bae7cfa0aedd4b4422d8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 11:33:59 +0100 Subject: [PATCH 034/149] Add deeptools --- environment.yml | 3 +++ 1 file changed, 3 insertions(+) diff --git a/environment.yml b/environment.yml index dbffed49..03b9307c 100755 --- a/environment.yml +++ b/environment.yml @@ -7,6 +7,7 @@ channels: - defaults dependencies: ## conda-forge packages + - gawk=4.2.1 - r-base=3.4.1 - r-optparse=1.6.0 - r-rcolorbrewer=1.1_2 @@ -16,6 +17,7 @@ dependencies: - r-pheatmap=1.0.10 - r-lattice=0.20_35 - r-upsetr=1.3.3 + - r-xfun=0.3 ## bioconda packages - fastqc=0.11.8 @@ -27,6 +29,7 @@ dependencies: - pysam=0.15.2 - bedtools=2.27.1 - preseq=2.0.3 + - deeptools=3.2.1 - ucsc-bedgraphtobigwig=377 - macs2=2.1.2 - homer=4.9.1 From ab6016593da41d5ce5969d5d78346101af5baad8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:25:22 +0100 Subject: [PATCH 035/149] Use min_reps_consensus param --- bin/igv_files_to_session.py | 13 +++++++++---- 1 file changed, 9 insertions(+), 4 deletions(-) diff --git a/bin/igv_files_to_session.py b/bin/igv_files_to_session.py index b9537c05..6dc6aabe 100755 --- a/bin/igv_files_to_session.py +++ b/bin/igv_files_to_session.py @@ -24,6 +24,9 @@ argParser.add_argument('XML_OUT', help="XML output file.") argParser.add_argument('LIST_FILE', help="Tab-delimited file containing two columns i.e. file_name\tcolour. Header isnt required.") argParser.add_argument('GENOME', help="Full path to genome fasta file or shorthand for genome available in IGV e.g. hg19.") + +## OPTIONAL PARAMETERS +argParser.add_argument('-pp', '--path_prefix', type=str, dest="PATH_PREFIX", default='', help="Path prefix to be added at beginning of all files in input list file.") args = argParser.parse_args() ############################################ @@ -47,7 +50,7 @@ def makedir(path): ############################################ ############################################ -def igv_files_to_session(XMLOut,ListFile,Genome): +def igv_files_to_session(XMLOut,ListFile,Genome,PathPrefix=''): makedir(os.path.dirname(XMLOut)) @@ -57,7 +60,9 @@ def igv_files_to_session(XMLOut,ListFile,Genome): line = fin.readline() if line: ifile,colour = line.strip().split('\t') - fileList.append((ifile,colour)) + if len(colour.strip()) == 0: + colour = '0,0,178' + fileList.append((PathPrefix.strip()+ifile,colour)) else: break fout.close() @@ -74,7 +79,7 @@ def igv_files_to_session(XMLOut,ListFile,Genome): XMLStr += '\t\n' for ifile,colour in fileList: extension = os.path.splitext(ifile)[1].lower() - if extension in ['.bed']: + if extension in ['.bed','.broadpeak','.narrowpeak']: XMLStr += '\t\t Date: Thu, 3 Oct 2019 13:25:45 +0100 Subject: [PATCH 036/149] Change file staging for igv --- bin/igv_files_to_session.py | 121 ------------------------------------ 1 file changed, 121 deletions(-) delete mode 100755 bin/igv_files_to_session.py diff --git a/bin/igv_files_to_session.py b/bin/igv_files_to_session.py deleted file mode 100755 index 6dc6aabe..00000000 --- a/bin/igv_files_to_session.py +++ /dev/null @@ -1,121 +0,0 @@ -#!/usr/bin/env python - -####################################################################### -####################################################################### -## Created on July 4th 2018 to create IGV session file from file list -####################################################################### -####################################################################### - -import os -import argparse - -############################################ -############################################ -## PARSE ARGUMENTS -############################################ -############################################ - -Description = 'Create IGV session file from a list of files and associated colours - ".bed", ".bw", ".bigwig", ".tdf", ".gtf" files currently supported.' -Epilog = """Example usage: python igv_files_to_session.py """ - -argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) - -## REQUIRED PARAMETERS -argParser.add_argument('XML_OUT', help="XML output file.") -argParser.add_argument('LIST_FILE', help="Tab-delimited file containing two columns i.e. file_name\tcolour. Header isnt required.") -argParser.add_argument('GENOME', help="Full path to genome fasta file or shorthand for genome available in IGV e.g. hg19.") - -## OPTIONAL PARAMETERS -argParser.add_argument('-pp', '--path_prefix', type=str, dest="PATH_PREFIX", default='', help="Path prefix to be added at beginning of all files in input list file.") -args = argParser.parse_args() - -############################################ -############################################ -## HELPER FUNCTIONS -############################################ -############################################ - -def makedir(path): - - if not len(path) == 0: - try: - os.makedirs(path) - except OSError as exception: - if exception.errno != errno.EEXIST: - raise - -############################################ -############################################ -## MAIN FUNCTION -############################################ -############################################ - -def igv_files_to_session(XMLOut,ListFile,Genome,PathPrefix=''): - - makedir(os.path.dirname(XMLOut)) - - fileList = [] - fin = open(ListFile,'r') - while True: - line = fin.readline() - if line: - ifile,colour = line.strip().split('\t') - if len(colour.strip()) == 0: - colour = '0,0,178' - fileList.append((PathPrefix.strip()+ifile,colour)) - else: - break - fout.close() - - ## ADD RESOURCES SECTION - XMLStr = '\n' - XMLStr += '\n' % (Genome) - XMLStr += '\t\n' - for ifile,colour in fileList: - XMLStr += '\t\t\n' % (ifile) - XMLStr += '\t\n' - - ## ADD PANEL SECTION - XMLStr += '\t\n' - for ifile,colour in fileList: - extension = os.path.splitext(ifile)[1].lower() - if extension in ['.bed','.broadpeak','.narrowpeak']: - XMLStr += '\t\t Date: Thu, 3 Oct 2019 13:26:18 +0100 Subject: [PATCH 037/149] Change file staging for igv --- bin/igv_get_files.sh | 22 ---------------------- 1 file changed, 22 deletions(-) delete mode 100755 bin/igv_get_files.sh diff --git a/bin/igv_get_files.sh b/bin/igv_get_files.sh deleted file mode 100755 index de1d81d4..00000000 --- a/bin/igv_get_files.sh +++ /dev/null @@ -1,22 +0,0 @@ -#!/usr/bin/env bash - -## ARGUMENTS -INPUT_DIR=$1 ## Path to input directory -FILE_GROUP=$2 ## File group identified -FILE_PATH=$3 ## Path to results/ directory containing files - -## CONSENSUS PEAKS -find -L $INPUT_DIR -type f -iname "consensus*$FILE_GROUP*" | cut -c 2- | awk -v DIR="$FILE_PATH" -v OFS='\t' '{ print DIR$1, "0,0,0" }' - -## DIFFERENTIAL INTERVALS FOR FDR <= 0.01 -for fpath in $(find -L $INPUT_DIR -type f -iname "*$FILE_GROUP*" | grep "FDR" | grep -Ev "FDR0.01" | sort | cut -c 2-) - do - fdir=$(dirname "${fpath}") - fbase=$(basename "${fpath}") - odir=$(echo "${fbase}" | awk -v var="."$FILE_GROUP 'BEGIN {FS=var;}{print $1}') - opath=$FILE_PATH/$fdir/$odir/$fbase - awk -v var="$opath" -v OFS='\t' 'BEGIN {print var, "255,0,0"}' - done - -## PEAK AND BIGWIG FILES -find -L $INPUT_DIR -type f -iname "*$FILE_GROUP*" | grep -Ev "consensus|FDR" | sort | cut -c 2- | awk -v DIR="$FILE_PATH" -v OFS='\t' '{ print DIR$1, "0,0,178" }' From ab535e2d98a73d4cd2531ea1d0a56a3446add701 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:49:25 +0100 Subject: [PATCH 038/149] Descibe additional processes --- README.md | 18 ++++++++++-------- 1 file changed, 10 insertions(+), 8 deletions(-) diff --git a/README.md b/README.md index 9de52ed2..391ffa41 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,3 @@ - # ![nfcore/atacseq](docs/images/nfcore-atacseq_logo.png) [![Build Status](https://travis-ci.org/nf-core/atacseq.svg?branch=master)](https://travis-ci.org/nf-core/atacseq) @@ -35,13 +34,16 @@ The pipeline is built using [Nextflow](https://www.nextflow.io), a workflow tool * reads that map to different chromosomes ([`Pysam`](http://pysam.readthedocs.io/en/latest/installation.html); *paired-end only*) * reads that arent in FR orientation ([`Pysam`](http://pysam.readthedocs.io/en/latest/installation.html); *paired-end only*) * reads where only one read of the pair fails the above criteria ([`Pysam`](http://pysam.readthedocs.io/en/latest/installation.html); *paired-end only*) - 3. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`wigToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) - 4. Call broad/narrow peaks ([`MACS2`](https://github.com/taoliu/MACS)) - 5. Annotate peaks relative to gene features ([`HOMER`](http://homer.ucsd.edu/homer/download.html)) - 6. Create consensus peakset across all samples and create tabular file to aid in the filtering of the data ([`BEDTools`](https://github.com/arq5x/bedtools2/)) - 7. Count reads in consensus peaks ([`featureCounts`](http://bioinf.wehi.edu.au/featureCounts/)) - 8. Differential accessibility analysis, PCA and clustering ([`R`](https://www.r-project.org/), [`DESeq2`](https://bioconductor.org/packages/release/bioc/html/DESeq2.html)) - 9. Generate ATAC-seq specific QC html report ([`ataqv`](https://github.com/ParkerLab/ataqv)) + 3. Alignment-level QC and estimation of library complexity ([`picard`](https://broadinstitute.github.io/picard/), [`Preseq`](http://smithlabresearch.org/software/preseq/)) + 4. Create normalised bigWig files scaled to 1 million mapped reads ([`BEDTools`](https://github.com/arq5x/bedtools2/), [`wigToBigWig`](http://hgdownload.soe.ucsc.edu/admin/exe/)) + 5. Generate gene-body meta-profile from bigWig files ([`deepTools`](https://deeptools.readthedocs.io/en/develop/content/tools/plotProfile.html)) + 6. Calculate genome-wide enrichment ([`deepTools`](https://deeptools.readthedocs.io/en/develop/content/tools/plotFingerprint.html)) + 7. Call broad/narrow peaks ([`MACS2`](https://github.com/taoliu/MACS)) + 8. Annotate peaks relative to gene features ([`HOMER`](http://homer.ucsd.edu/homer/download.html)) + 9. Create consensus peakset across all samples and create tabular file to aid in the filtering of the data ([`BEDTools`](https://github.com/arq5x/bedtools2/)) + 10. Count reads in consensus peaks ([`featureCounts`](http://bioinf.wehi.edu.au/featureCounts/)) + 11. Differential accessibility analysis, PCA and clustering ([`R`](https://www.r-project.org/), [`DESeq2`](https://bioconductor.org/packages/release/bioc/html/DESeq2.html)) + 12. Generate ATAC-seq specific QC html report ([`ataqv`](https://github.com/ParkerLab/ataqv)) 6. Merge filtered alignments across replicates ([`picard`](https://broadinstitute.github.io/picard/)) 1. Re-mark duplicates ([`picard`](https://broadinstitute.github.io/picard/)) 2. Remove duplicate reads ([`SAMtools`](https://sourceforge.net/projects/samtools/files/samtools/)) From c0413a3a2592f3ba9e6bda3f7319516a0ea780b7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:49:57 +0100 Subject: [PATCH 039/149] Update scripts in bin directory --- bin/check_design.py | 8 ++-- bin/macs2_merged_expand.py | 85 +++++++++++++++++++-------------- bin/plot_homer_annotatepeaks.r | 2 +- bin/scrape_software_versions.py | 4 ++ 4 files changed, 58 insertions(+), 41 deletions(-) diff --git a/bin/check_design.py b/bin/check_design.py index 8647b0c7..4c1436f0 100755 --- a/bin/check_design.py +++ b/bin/check_design.py @@ -2,7 +2,7 @@ ####################################################################### ####################################################################### -## Created on February 22nd 2019 to reformat nf-core/atacseq design file +## Created on February 22nd 2019 to check nf-core/atacseq design file ####################################################################### ####################################################################### @@ -18,7 +18,7 @@ ############################################ Description = 'Reformat nf-core/atacseq design file and check its contents.' -Epilog = """Example usage: python reformat_design.py """ +Epilog = """Example usage: python check_design.py """ argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) @@ -33,7 +33,7 @@ ############################################ ############################################ -def reformat_design(DesignFileIn,DesignFileOut): +def check_design(DesignFileIn,DesignFileOut): ERROR_STR = 'ERROR: Please check design file' HEADER = ['group', 'replicate', 'fastq_1', 'fastq_2'] @@ -137,7 +137,7 @@ def reformat_design(DesignFileIn,DesignFileOut): ############################################ ############################################ -reformat_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) +check_design(DesignFileIn=args.DESIGN_FILE_IN,DesignFileOut=args.DESIGN_FILE_OUT) ############################################ ############################################ diff --git a/bin/macs2_merged_expand.py b/bin/macs2_merged_expand.py index 41f98578..10889947 100755 --- a/bin/macs2_merged_expand.py +++ b/bin/macs2_merged_expand.py @@ -16,18 +16,18 @@ ############################################ Description = 'Add sample boolean files and aggregate columns from merged MACS narrow or broad peak file.' -Epilog = """Example usage: python macs2_merged_expand.py --is_narrow_peak --min_samples 0""" +Epilog = """Example usage: python macs2_merged_expand.py --is_narrow_peak --min_replicates 1""" argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) ## REQUIRED PARAMETERS argParser.add_argument('MERGED_INTERVAL_FILE', help="Merged MACS2 interval file created using linux sort and mergeBed.") -argParser.add_argument('SAMPLE_NAME_LIST', help="Comma-separated list of sample names as named in individual MACS2 broadPeak/narrowPeak output file e.g. SAMPLE_1 for SAMPLE_1_peak_1.") +argParser.add_argument('SAMPLE_NAME_LIST', help="Comma-separated list of sample names as named in individual MACS2 broadPeak/narrowPeak output file e.g. SAMPLE_R1 for SAMPLE_R1_peak_1.") argParser.add_argument('OUTFILE', help="Full path to output directory.") ## OPTIONAL PARAMETERS argParser.add_argument('-in', '--is_narrow_peak', dest="IS_NARROW_PEAK", help="Whether merged interval file was generated from narrow or broad peak files (default: False).",action='store_true') -argParser.add_argument('-ms', '--min_samples', type=int, dest="MIN_SAMPLES", default=0, help="Minumum number of samples required per merged peak (default: 0).") +argParser.add_argument('-mr', '--min_replicates', type=int, dest="MIN_REPLICATES", default=1, help="Minumum number of replicates per sample required to contribute to merged peak (default: 1).") args = argParser.parse_args() ############################################ @@ -57,13 +57,13 @@ def makedir(path): ## 2) narrowPeak ## sort -k1,1 -k2,2n | mergeBed -c 2,3,4,5,6,7,8,9,10 -o collapse,collapse,collapse,collapse,collapse,collapse,collapse,collapse,collapse > merged_peaks.txt -def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=False,minSamples=0): +def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=False,minReplicates=1): makedir(os.path.dirname(OutFile)) + combFreqDict = {} + totalOutIntervals = 0 SampleNameList = sorted(SampleNameList) - totalInIntervals = 0; totalOutIntervals = 0 - sampleFreqDict = {}; combFreqDict = {} fin = open(MergedIntervalTxtFile,'r') fout = open(OutFile,'w') oFields = ['chr','start','end','interval_id','num_peaks','num_samples'] + [x+'.bool' for x in SampleNameList] + [x+'.fc' for x in SampleNameList] + [x+'.qval' for x in SampleNameList] + [x+'.pval' for x in SampleNameList] + [x+'.start' for x in SampleNameList] + [x+'.end' for x in SampleNameList] @@ -83,32 +83,49 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa if isNarrow: summits = [int(x) for x in lspl[11].split(',')] + ## GROUP SAMPLES BY REMOVING TRAILING *_R* + groupDict = {} + for sID in ['_'.join(x.split('_')[:-2]) for x in names]: + gID = '_'.join(sID.split('_')[:-1]) + if not groupDict.has_key(gID): + groupDict[gID] = [] + if not sID in groupDict[gID]: + groupDict[gID].append(sID) + + ## GET SAMPLES THAT PASS REPLICATE THRESHOLD + passRepThreshList = [] + for gID,sIDs in groupDict.items(): + if len(sIDs) >= minReplicates: + passRepThreshList += sIDs + + ## GET VALUES FROM INDIVIDUAL PEAK SETS fcDict = {}; qvalDict = {}; pvalDict = {}; startDict = {}; endDict = {}; summitDict = {} for idx in range(len(names)): sample = '_'.join(names[idx].split('_')[:-2]) - if not fcDict.has_key(sample): - fcDict[sample] = [] - fcDict[sample].append(str(fcs[idx])) - if not qvalDict.has_key(sample): - qvalDict[sample] = [] - qvalDict[sample].append(str(qvals[idx])) - if not pvalDict.has_key(sample): - pvalDict[sample] = [] - pvalDict[sample].append(str(pvals[idx])) - if not startDict.has_key(sample): - startDict[sample] = [] - startDict[sample].append(str(starts[idx])) - if not endDict.has_key(sample): - endDict[sample] = [] - endDict[sample].append(str(ends[idx])) - if isNarrow: - if not summitDict.has_key(sample): - summitDict[sample] = [] - summitDict[sample].append(str(summits[idx])) + if sample in passRepThreshList: + if not fcDict.has_key(sample): + fcDict[sample] = [] + fcDict[sample].append(str(fcs[idx])) + if not qvalDict.has_key(sample): + qvalDict[sample] = [] + qvalDict[sample].append(str(qvals[idx])) + if not pvalDict.has_key(sample): + pvalDict[sample] = [] + pvalDict[sample].append(str(pvals[idx])) + if not startDict.has_key(sample): + startDict[sample] = [] + startDict[sample].append(str(starts[idx])) + if not endDict.has_key(sample): + endDict[sample] = [] + endDict[sample].append(str(ends[idx])) + if isNarrow: + if not summitDict.has_key(sample): + summitDict[sample] = [] + summitDict[sample].append(str(summits[idx])) samples = sorted(fcDict.keys()) - numSamples = len(samples) - if numSamples >= minSamples: + if samples != []: + numSamples = len(samples) boolList = ['TRUE' if x in samples else 'FALSE' for x in SampleNameList] fcList = [';'.join(fcDict[x]) if x in samples else 'NA' for x in SampleNameList] qvalList = [';'.join(qvalDict[x]) if x in samples else 'NA' for x in SampleNameList] @@ -120,16 +137,12 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa oList += [';'.join(summitDict[x]) if x in samples else 'NA' for x in SampleNameList] fout.write('\t'.join(oList) + '\n') - if not sampleFreqDict.has_key(numSamples): - sampleFreqDict[numSamples] = 0 - sampleFreqDict[numSamples] += 1 + tsamples = tuple(sorted(samples)) + if not combFreqDict.has_key(tsamples): + combFreqDict[tsamples] = 0 + combFreqDict[tsamples] += 1 totalOutIntervals += 1 - tsamples = tuple(sorted(samples)) - if not combFreqDict.has_key(tsamples): - combFreqDict[tsamples] = 0 - combFreqDict[tsamples] += 1 - totalInIntervals += 1 else: fin.close() fout.close() @@ -149,7 +162,7 @@ def macs2_merged_expand(MergedIntervalTxtFile,SampleNameList,OutFile,isNarrow=Fa ############################################ ############################################ -macs2_merged_expand(MergedIntervalTxtFile=args.MERGED_INTERVAL_FILE,SampleNameList=args.SAMPLE_NAME_LIST.split(','),OutFile=args.OUTFILE,isNarrow=args.IS_NARROW_PEAK,minSamples=args.MIN_SAMPLES) +macs2_merged_expand(MergedIntervalTxtFile=args.MERGED_INTERVAL_FILE,SampleNameList=args.SAMPLE_NAME_LIST.split(','),OutFile=args.OUTFILE,isNarrow=args.IS_NARROW_PEAK,minReplicates=args.MIN_REPLICATES) ############################################ ############################################ diff --git a/bin/plot_homer_annotatepeaks.r b/bin/plot_homer_annotatepeaks.r index 1eb0ec3c..ee537fce 100755 --- a/bin/plot_homer_annotatepeaks.r +++ b/bin/plot_homer_annotatepeaks.r @@ -57,7 +57,7 @@ plot.feature.dat <- data.frame() for (idx in 1:length(HomerFiles)) { sampleid = SampleIDs[idx] - anno.dat <- read.table(HomerFiles[idx], sep="\t", header=TRUE) + anno.dat <- read.table(HomerFiles[idx], sep="\t", header=TRUE,quote="") anno.dat <- anno.dat[,c("Annotation","Distance.to.TSS","Nearest.PromoterID")] anno.dat <- anno.dat[which(!is.na(anno.dat$Distance.to.TSS)),] if (nrow(anno.dat) == 0) { diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index 22cc5458..456c8fae 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -12,6 +12,7 @@ 'Samtools': ['v_samtools.txt', r"samtools (\S+)"], 'BEDTools': ['v_bedtools.txt', r"bedtools v(\S+)"], 'BamTools': ['v_bamtools.txt', r"bamtools (\S+)"], + #'deepTools': ['v_deeptools.txt', r"plotFingerprint (\S+)"], 'Picard': ['v_picard.txt', r"([\d\.]+)-SNAPSHOT"], 'R': ['v_R.txt', r"R version (\S+)"], 'Pysam': ['v_pysam.txt', r"(\S+)"], @@ -19,6 +20,7 @@ 'HOMER': ['v_homer.txt', r"(\S+)"], 'ataqv': ['v_ataqv.txt', r"(\S+)"], 'featureCounts': ['v_featurecounts.txt', r"featureCounts v(\S+)"], + #'Preseq': ['v_preseq.txt', r"Version: (\S+)"], 'MultiQC': ['v_multiqc.txt', r"multiqc, version (\S+)"], } @@ -31,6 +33,7 @@ results['Samtools'] = 'N/A' results['BEDTools'] = 'N/A' results['BamTools'] = 'N/A' +#results['deepTools'] = 'N/A' results['Picard'] = 'N/A' results['R'] = 'N/A' results['Pysam'] = 'N/A' @@ -38,6 +41,7 @@ results['HOMER'] = False results['ataqv'] = 'N/A' results['featureCounts'] = 'N/A' +#results['Preseq'] = 'N/A' results['MultiQC'] = 'N/A' # Search each file using its regex From bdbe090feb2c03c3ee9d6d609a02292db8a802d9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:50:13 +0100 Subject: [PATCH 040/149] Change process_log --- conf/base.config | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/conf/base.config b/conf/base.config index 9d01790c..48ac17fa 100755 --- a/conf/base.config +++ b/conf/base.config @@ -36,7 +36,7 @@ process { time = { check_max( 16.h * task.attempt, 'time' ) } } withLabel:process_long { - time = { check_max( 8.h * task.attempt, 'time' ) } + time = { check_max( 12.h * task.attempt, 'time' ) } } withName:get_software_versions { cache = false From e8a90fdb6bca1740587fc6421ead2f35f78962a8 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:50:32 +0100 Subject: [PATCH 041/149] Add fingerprint bins --- conf/test.config | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/conf/test.config b/conf/test.config index 9ed9db45..b8fb3eee 100755 --- a/conf/test.config +++ b/conf/test.config @@ -19,10 +19,13 @@ params { design = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/design.csv' // Genome references + mito_name = 'MT' fasta = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genome.fa' gtf = 'https://raw.githubusercontent.com/nf-core/test-datasets/atacseq/reference/genes.gtf' // Not mandatory but permits the pipeline to run through peak-calling steps - mito_name = 'MT' macs_gsize = 1.2e7 + + // For speed to avoid CI time-out + fingerprint_bins = 100 } From 94c5a3d54fce9279de049913a71fdd2808b80244 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 13:58:26 +0100 Subject: [PATCH 042/149] Update params with chipseq --- nextflow.config | 26 +++++++++++++++++++++----- 1 file changed, 21 insertions(+), 5 deletions(-) diff --git a/nextflow.config b/nextflow.config index cc0f62c4..df68009a 100755 --- a/nextflow.config +++ b/nextflow.config @@ -9,12 +9,13 @@ params { // Options: Generic - genome = false singleEnd = false - narrowPeak = false + seq_center = false fragment_size = 0 + fingerprint_bins = 500000 // Options: References + genome = false tss_bed = false saveGenomeIndex = false @@ -33,6 +34,23 @@ params { skipMergeReplicates = false saveAlignedIntermediates = false + // Options: Peaks + narrowPeak = false + broad_cutoff = 0.1 + min_reps_consensus = 1 + saveMACSPileup = false + skipDiffAnalysis = false + + // Options: QC + skipFastQC = false + skipPicardMetrics = false + skipPreseq = false + skipPlotProfile = false + skipPlotFingerprint = false + skipIGV = false + skipMultiQC = false + skipMultiQCStats = false + // Options: Other outdir = './results' igenomes_base = "./iGenomes" @@ -96,9 +114,7 @@ if(!params.igenomesIgnore){ } // Increase time available to build conda environment -conda { - createTimeout = "60 min" -} +conda { createTimeout = "60 min" } // Capture exit codes from upstream processes when piping process.shell = ['/bin/bash', '-euo', 'pipefail'] From 3a76775dcc2b0b3867cc434690b36e2f515e441e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 18:24:56 +0100 Subject: [PATCH 043/149] Major overhaul of channels and processes --- main.nf | 1426 ++++++++++++++++++++++++++++++++----------------------- 1 file changed, 829 insertions(+), 597 deletions(-) diff --git a/main.nf b/main.nf index 0904ad61..f7bde6c8 100644 --- a/main.nf +++ b/main.nf @@ -27,43 +27,59 @@ def helpMessage() { nextflow run nf-core/atacseq --design design.csv --genome GRCh37 -profile docker Mandatory arguments: - --design Comma-separted file containing information about the samples in the experiment (see docs/usage.md) + --design Comma-separated file containing information about the samples in the experiment (see docs/usage.md) --fasta Path to Fasta reference. Not mandatory when using reference in iGenomes config via --genome - --gtf Path to GTF file in Ensembl format. Not mandatory when using reference in iGenomes config via --genome + --gtf Path to GTF file. Not mandatory when using reference in iGenomes config via --genome -profile Configuration profile to use. Can use multiple (comma separated) Available: conda, docker, singularity, awsbatch, test Generic - --genome Name of iGenomes reference --singleEnd Specifies that the input is single-end reads - --narrowPeak Run MACS in narrowPeak mode. Default: broadPeak - --fragment_size [int] Estimated fragment size used to extend single-end reads. Default: 0 + --seq_center Sequencing center information to be added to read group of BAM files + --fragment_size [int] Estimated fragment size used to extend single-end reads (Default: 0) + --fingerprint_bins [int] Number of genomic bins to use when calculating fingerprint plot (Default: 500000) References If not specified in the configuration file or you wish to overwrite any of the references + --genome Name of iGenomes reference --bwa_index Full path to directory containing BWA index including base name i.e. /path/to/index/genome.fa --gene_bed Path to BED file containing gene intervals - --tss_bed Path to BED file containing transcription start sites (used by ataqv) - --mito_name Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out - + --tss_bed Path to BED file containing transcription start sites --macs_gsize Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235 --blacklist Path to blacklist regions (.BED format), used for filtering alignments + --mito_name Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out --saveGenomeIndex If generated by the pipeline save the BWA index in the results directory Trimming - --clip_r1 [int] Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads) - --clip_r2 [int] Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only) - --three_prime_clip_r1 [int] Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed - --three_prime_clip_r2 [int] Instructs Trim Galore to re move bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed + --clip_r1 [int] Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads) (Default: 0) + --clip_r2 [int] Instructs Trim Galore to remove bp from the 5' end of read 2 (paired-end reads only) (Default: 0) + --three_prime_clip_r1 [int] Instructs Trim Galore to remove bp from the 3' end of read 1 AFTER adapter/quality trimming has been performed (Default: 0) + --three_prime_clip_r2 [int] Instructs Trim Galore to re move bp from the 3' end of read 2 AFTER adapter/quality trimming has been performed (Default: 0) --skipTrimming Skip the adapter trimming step - --saveTrimmed Save the trimmed FastQ files in the the Results directory + --saveTrimmed Save the trimmed FastQ files in the results directory Alignments --keepMito Reads mapping to mitochondrial contig are not filtered from alignments --keepDups Duplicate reads are not filtered from alignments --keepMultiMap Reads mapping to multiple locations are not filtered from alignments - --skipMergeReplicates Do not perform alignment merging and downstream analysis by merging replicates i.e. only do this by merging resequenced libraries --saveAlignedIntermediates Save the intermediate BAM files from the alignment step - not done by default + Peaks + --narrowPeak Run MACS2 in narrowPeak mode + --broad_cutoff [float] Specifies broad cutoff value for MACS2. Only used when --narrowPeak isnt specified (Default: 0.1) + --min_reps_consensus Number of biological replicates required from a given condition for a peak to contribute to a consensus peak (Default: 1) + --saveMACSPileup Instruct MACS2 to create bedGraph files normalised to signal per million reads + --skipDiffAnalysis Skip differential binding analysis + + QC + --skipFastQC Skip FastQC + --skipPicardMetrics Skip Picard CollectMultipleMetrics + --skipPreseq Skip Preseq + --skipPlotProfile Skip deepTools plotProfile + --skipPlotFingerprint Skip deepTools plotFingerprint + --skipIGV Skip IGV + --skipMultiQC Skip MultiQC + --skipMultiQCStats Exclude general statistics table from MultiQC report + Other --outdir The output directory where the results will be saved --email Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits @@ -89,7 +105,7 @@ def helpMessage() { */ // Show help message -if (params.help){ +if (params.help) { helpMessage() exit 0 } @@ -115,8 +131,8 @@ params.blacklist = params.genome ? params.genomes[ params.genome ].blacklist ?: // Has the run name been specified by the user? // this has the bonus effect of catching both -name and --name custom_runName = params.name -if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)){ - custom_runName = workflow.runName +if (!(workflow.runName ==~ /[a-z]+_[a-z]+/)) { + custom_runName = workflow.runName } //////////////////////////////////////////////////// @@ -158,7 +174,7 @@ if (params.gene_bed) { ch_gene_bed = file(params.gene.bed, checkIfExists: true) if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } -if (params.fasta){ +if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) bwa_base = params.fasta.substring(lastPath+1) ch_fasta = file(params.fasta, checkIfExists: true) @@ -166,7 +182,7 @@ if (params.fasta){ exit 1, "Fasta file not specified!" } -if (params.bwa_index){ +if (params.bwa_index) { lastPath = params.bwa_index.lastIndexOf(File.separator) bwa_dir = params.bwa_index.substring(0,lastPath+1) bwa_base = params.bwa_index.substring(lastPath+1) @@ -178,14 +194,14 @@ if (params.bwa_index){ /* -- AWS -- */ //////////////////////////////////////////////////// -if( workflow.profile == 'awsbatch') { - // AWSBatch sanity checking - if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" - // Check outdir paths to be S3 buckets if running on AWSBatch - // related: https://github.com/nextflow-io/nextflow/issues/813 - if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" - // Prevent trace files to be stored on S3 since S3 does not support rolling files. - if (workflow.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." +if (workflow.profile == 'awsbatch') { + // AWSBatch sanity checking + if (!params.awsqueue || !params.awsregion) exit 1, "Specify correct --awsqueue and --awsregion parameters on AWSBatch!" + // Check outdir paths to be S3 buckets if running on AWSBatch + // related: https://github.com/nextflow-io/nextflow/issues/813 + if (!params.outdir.startsWith('s3:')) exit 1, "Outdir not on S3 - specify S3 Bucket to run on AWSBatch!" + // Prevent trace files to be stored on S3 since S3 does not support rolling files. + if (workflow.tracedir.startsWith('s3:')) exit 1, "Specify a local tracedir or run without trace! S3 cannot be used for tracefiles." } /////////////////////////////////////////////////////////////////////////////// @@ -200,19 +216,21 @@ if( workflow.profile == 'awsbatch') { log.info nfcoreHeader() def summary = [:] summary['Run Name'] = custom_runName ?: workflow.runName -summary['Genome'] = params.genome ?: 'Not supplied' summary['Data Type'] = params.singleEnd ? 'Single-End' : 'Paired-End' summary['Design File'] = params.design -if (params.bwa_index) summary['BWA Index'] = params.bwa_index ?: 'Not supplied' -summary['Fasta Ref'] = params.fasta +summary['Genome'] = params.genome ?: 'Not supplied' +summary['Fasta File'] = params.fasta summary['GTF File'] = params.gtf -summary['Gene BED File'] = params.gene_bed ?: 'Not supplied' -summary['TSS BED File'] = params.tss_bed ?: 'Not supplied' -if (params.blacklist) summary['Blacklist BED'] = params.blacklist -summary['Mitochondrial Contig'] = params.mito_name ?: 'Not supplied' -summary['MACS Genome Size'] = params.macs_gsize ?: 'Not supplied' -if (params.macs_gsize) summary['MACS Narrow Peaks'] = params.narrowPeak ? 'Yes' : 'No' -if (params.skipTrimming){ +if (params.gene_bed) summary['Gene BED File'] = params.gene_bed +if (params.tss_bed) summary['TSS BED File'] = params.tss_bed +if (params.bwa_index) summary['BWA Index'] = params.bwa_index +if (params.blacklist) summary['Blacklist BED'] = params.blacklist +if (params.mito_name) summary['Mitochondrial Contig'] = params.mito_name +summary['MACS2 Genome Size'] = params.macs_gsize ?: 'Not supplied' +summary['Min Consensus Reps'] = params.min_reps_consensus +if (params.macs_gsize) summary['MACS2 Narrow Peaks'] = params.narrowPeak ? 'Yes' : 'No' +if (!params.narrowPeak) summary['MACS2 Broad Cutoff'] = params.broad_cutoff +if (params.skipTrimming) { summary['Trimming Step'] = 'Skipped' } else { summary['Trim R1'] = "$params.clip_r1 bp" @@ -220,32 +238,44 @@ if (params.skipTrimming){ summary["Trim 3' R1"] = "$params.three_prime_clip_r1 bp" summary["Trim 3' R2"] = "$params.three_prime_clip_r2 bp" } -summary['Fragment Size'] = "$params.fragment_size bp" -summary['Keep Mitochondrial'] = params.keepMito ? 'Yes' : 'No' -summary['Keep Duplicates'] = params.keepDups ? 'Yes' : 'No' -summary['Keep Multi-mapped'] = params.keepMultiMap ? 'Yes' : 'No' -summary['Merge Replicates'] = params.skipMergeReplicates ? 'No' : 'Yes' +if (params.seq_center) summary['Sequencing Center'] = params.seq_center +if (params.singleEnd) summary['Fragment Size'] = "$params.fragment_size bp" +summary['Fingerprint Bins'] = params.fingerprint_bins +if (params.keepMito) summary['Keep Mitochondrial'] = 'Yes' +if (params.keepDups) summary['Keep Duplicates'] = 'Yes' +if (params.keepMultiMap) summary['Keep Multi-mapped'] = 'Yes' summary['Save Genome Index'] = params.saveGenomeIndex ? 'Yes' : 'No' -summary['Save Trimmed'] = params.saveTrimmed ? 'Yes' : 'No' -summary['Save Intermeds'] = params.saveAlignedIntermediates ? 'Yes' : 'No' +if (params.saveTrimmed) summary['Save Trimmed'] = 'Yes' +if (params.saveAlignedIntermediates) summary['Save Intermeds'] = 'Yes' +if (params.saveMACSPileup) summary['Save MACS2 Pileup'] = 'Yes' +if (params.skipMergeReplicates) summary['Skip Merge Replicates'] = 'Yes' +if (params.skipDiffAnalysis) summary['Skip Diff Analysis'] = 'Yes' +if (params.skipFastQC) summary['Skip FastQC'] = 'Yes' +if (params.skipPicardMetrics) summary['Skip Picard Metrics'] = 'Yes' +if (params.skipPreseq) summary['Skip Preseq'] = 'Yes' +if (params.skipPlotProfile) summary['Skip plotProfile'] = 'Yes' +if (params.skipPlotFingerprint) summary['Skip plotFingerprint'] = 'Yes' +if (params.skipIGV) summary['Skip IGV'] = 'Yes' +if (params.skipMultiQC) summary['Skip MultiQC'] = 'Yes' +if (params.skipMultiQCStats) summary['Skip MultiQC Stats'] = 'Yes' summary['Max Resources'] = "$params.max_memory memory, $params.max_cpus cpus, $params.max_time time per job" -if(workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" +if (workflow.containerEngine) summary['Container'] = "$workflow.containerEngine - $workflow.container" summary['Output Dir'] = params.outdir summary['Launch Dir'] = workflow.launchDir summary['Working Dir'] = workflow.workDir summary['Script Dir'] = workflow.projectDir summary['User'] = workflow.userName -if (workflow.profile == 'awsbatch'){ - summary['AWS Region'] = params.awsregion - summary['AWS Queue'] = params.awsqueue +if (workflow.profile == 'awsbatch') { + summary['AWS Region'] = params.awsregion + summary['AWS Queue'] = params.awsqueue } summary['Config Profile'] = workflow.profile if (params.config_profile_description) summary['Config Description'] = params.config_profile_description if (params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact if (params.config_profile_url) summary['Config URL'] = params.config_profile_url -if(params.email) { - summary['E-mail Address'] = params.email - summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize +if (params.email) { + summary['E-mail Address'] = params.email + summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize } log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join("\n") log.info "\033[2m----------------------------------------------------\033[0m" @@ -254,7 +284,7 @@ log.info "\033[2m----------------------------------------------------\033[0m" checkHostname() // Show a big warning message if we're not running MACS -if (!params.macs_gsize){ +if (!params.macs_gsize) { def warnstring = params.genome ? "supported for '${params.genome}'" : 'supplied' log.warn "=================================================================\n" + " WARNING! MACS genome size parameter not $warnstring.\n" + @@ -295,32 +325,32 @@ process CheckDesign { */ if (params.singleEnd) { ch_design_reads_csv.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + .map { row -> [ row.sample_id, [ file(row.fastq_1, checkIfExists: true) ] ] } + .into { ch_raw_reads_fastqc; + ch_raw_reads_trimgalore; + design_replicates_exist; + design_multiple_samples } } else { ch_design_reads_csv.splitCsv(header:true, sep:',') - .map { row -> [ row.sample_id, [ file(row.fastq_1), file(row.fastq_2) ] ] } - .into { design_replicates_exist; - design_multiple_samples; - raw_reads_fastqc; - raw_reads_trimgalore } + .map { row -> [ row.sample_id, [ file(row.fastq_1, checkIfExists: true), file(row.fastq_2, checkIfExists: true) ] ] } + .into { ch_raw_reads_fastqc; + ch_raw_reads_trimgalore; + design_replicates_exist; + design_multiple_samples } } // Boolean value for replicates existing in design -replicates_exist = design_replicates_exist.map { it -> it[0].split('_')[-2].replaceAll('R','').toInteger() } - .flatten() - .max() - .val > 1 +replicatesExist = design_replicates_exist.map { it -> it[0].split('_')[-2].replaceAll('R','').toInteger() } + .flatten() + .max() + .val > 1 -// Boolean value for multiple samples existing in design -multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].join('_') } - .flatten() - .unique() - .count() - .val > 1 +// Boolean value for multiple groups existing in design +multipleGroups = design_multiple_samples.map { it -> it[0].split('_')[0..-3].join('_') } + .flatten() + .unique() + .count() + .val > 1 /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// @@ -333,12 +363,12 @@ multiple_samples = design_multiple_samples.map { it -> it[0].split('_')[0..-3].j /* * PREPROCESSING - Build BWA index */ -if (!params.bwa_index){ +if (!params.bwa_index) { process BWAIndex { tag "$fasta" label 'process_high' publishDir path: { params.saveGenomeIndex ? "${params.outdir}/reference_genome" : params.outdir }, - saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' + saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' input: file fasta from ch_fasta @@ -357,7 +387,7 @@ if (!params.bwa_index){ /* * PREPROCESSING - Generate gene BED file */ -if (!params.gene_bed){ +if (!params.gene_bed) { process MakeGeneBED { tag "$gtf" label 'process_low' @@ -379,7 +409,7 @@ if (!params.gene_bed){ /* * PREPROCESSING - Generate TSS BED file */ -if (!params.tss_bed){ +if (!params.tss_bed) { process MakeTSSBED { tag "$bed" publishDir "${params.outdir}/reference_genome", mode: 'copy' @@ -406,20 +436,20 @@ process MakeGenomeFilter { input: file fasta from ch_fasta - file blacklist from ch_blacklist + file blacklist from ch_blacklist.ifEmpty([]) output: - file "$fasta" into genome_fasta // FASTA FILE FOR IGV - file "*.fai" into genome_fai // FAI INDEX FOR REFERENCE GENOME - file "*.txt" into genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV - file "*.bed" into genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING - file "*.sizes" into genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS - genome_sizes_mrep_bigwig + file "$fasta" into ch_genome_fasta // FASTA FILE FOR IGV + file "*.fai" into ch_genome_fai // FAI INDEX FOR REFERENCE GENOME + file "*.bed" into ch_genome_filter_regions // BED FILE WITHOUT BLACKLIST REGIONS & MITOCHONDRIAL CONTIG FOR FILTERING + file "*.txt" into ch_genome_autosomes // TEXT FILE CONTAINING LISTING OF AUTOSOMAL CHROMOSOMES FOR ATAQV + file "*.sizes" into ch_genome_sizes_mlib_bigwig, // CHROMOSOME SIZES FILE FOR BEDTOOLS + ch_genome_sizes_mrep_bigwig script: - def blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" - def name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" - def mito_filter = params.keepMito ? "" : name_filter + blacklist_filter = params.blacklist ? "sortBed -i $blacklist -g ${fasta}.sizes | complementBed -i stdin -g ${fasta}.sizes" : "awk '{print \$1, '0' , \$2}' OFS='\t' ${fasta}.sizes" + name_filter = params.mito_name ? "| awk '\$1 !~ /${params.mito_name}/ {print \$0}'": "" + mito_filter = params.keepMito ? "" : name_filter """ samtools faidx $fasta get_autosomes.py ${fasta}.fai ${fasta}.autosomes.txt @@ -445,11 +475,14 @@ process FastQC { publishDir "${params.outdir}/fastqc", mode: 'copy', saveAs: {filename -> filename.endsWith(".zip") ? "zips/$filename" : "$filename"} + when: + !params.skipFastQC + input: - set val(name), file(reads) from raw_reads_fastqc + set val(name), file(reads) from ch_raw_reads_fastqc output: - file "*.{zip,html}" into fastqc_reports_mqc + file "*.{zip,html}" into ch_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC @@ -479,10 +512,10 @@ process FastQC { /* * STEP 2 - Trim Galore! */ -if (params.skipTrimming){ - trimmed_reads = raw_reads_trimgalore - trimgalore_results_mqc = [] - trimgalore_fastqc_reports_mqc = [] +if (params.skipTrimming) { + ch_trimmed_reads = ch_raw_reads_trimgalore + ch_trimgalore_results_mqc = [] + ch_trimgalore_fastqc_reports_mqc = [] } else { process TrimGalore { tag "$name" @@ -490,25 +523,25 @@ if (params.skipTrimming){ publishDir "${params.outdir}/trim_galore", mode: 'copy', saveAs: {filename -> if (filename.endsWith(".html")) "fastqc/$filename" - else if (filename.endsWith(".zip")) "fastqc/zip/$filename" + else if (filename.endsWith(".zip")) "fastqc/zips/$filename" else if (filename.endsWith("trimming_report.txt")) "logs/$filename" else params.saveTrimmed ? filename : null } input: - set val(name), file(reads) from raw_reads_trimgalore + set val(name), file(reads) from ch_raw_reads_trimgalore output: - set val(name), file("*.fq.gz") into trimmed_reads - file "*.txt" into trimgalore_results_mqc - file "*.{zip,html}" into trimgalore_fastqc_reports_mqc + set val(name), file("*.fq.gz") into ch_trimmed_reads + file "*.txt" into ch_trimgalore_results_mqc + file "*.{zip,html}" into ch_trimgalore_fastqc_reports_mqc script: // Added soft-links to original fastqs for consistent naming in MultiQC - def c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' - def c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' - def tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' - def tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' + c_r1 = params.clip_r1 > 0 ? "--clip_r1 ${params.clip_r1}" : '' + c_r2 = params.clip_r2 > 0 ? "--clip_r2 ${params.clip_r2}" : '' + tpc_r1 = params.three_prime_clip_r1 > 0 ? "--three_prime_clip_r1 ${params.three_prime_clip_r1}" : '' + tpc_r2 = params.three_prime_clip_r2 > 0 ? "--three_prime_clip_r2 ${params.three_prime_clip_r2}" : '' if (params.singleEnd) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz @@ -540,17 +573,26 @@ process BWAmem { label 'process_high' input: - set val(name), file(reads) from trimmed_reads + set val(name), file(reads) from ch_trimmed_reads file index from ch_bwa_index.collect() output: - set val(name), file("*.bam") into bwa_bam + set val(name), file("*.bam") into ch_bwa_bam script: - def prefix="${name}.Lb" - def rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + prefix = "${name}.Lb" + rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + if (params.seq_center) { + rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" + } """ - bwa mem -t $task.cpus -M -R $rg ${index}/${bwa_base} $reads | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - + bwa mem \\ + -t $task.cpus \\ + -M \\ + -R $rg \\ + ${index}/${bwa_base} \\ + $reads \\ + | samtools view -@ $task.cpus -b -h -F 0x0100 -O BAM -o ${prefix}.bam - """ } @@ -563,21 +605,21 @@ process SortBAM { if (params.saveAlignedIntermediates) { publishDir path: "${params.outdir}/bwa/library", mode: 'copy', saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else filename } - } + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else filename } + } input: - set val(name), file(bam) from bwa_bam + set val(name), file(bam) from ch_bwa_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into sort_bam_mlib - file "*.{flagstat,idxstats,stats}" into sort_bam_flagstat_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into ch_sort_bam_merge + file "*.{flagstat,idxstats,stats}" into ch_sort_bam_flagstat_mqc script: - def prefix="${name}.Lb" + prefix = "${name}.Lb" """ samtools sort -@ $task.cpus -o ${prefix}.sorted.bam -T $name $bam samtools index ${prefix}.sorted.bam @@ -598,12 +640,12 @@ process SortBAM { /* * STEP 4.1 Merge BAM files for all libraries from same replicate */ -sort_bam_mlib.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { sort_bam_mlib } +ch_sort_bam_merge.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_sort_bam_merge } -process MergeLibrary { +process MergeLibraryBAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -616,19 +658,20 @@ process MergeLibrary { } input: - set val(name), file(bams) from sort_bam_mlib + set val(name), file(bams) from ch_sort_bam_merge output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mlib_bam_filter, - mlib_bam_ataqv - file "*.{flagstat,idxstats,stats}" into mlib_bam_stats_mqc - file "*.txt" into mlib_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mlib_bam_filter, + ch_mlib_bam_preseq, + ch_mlib_bam_ataqv + file "*.{flagstat,idxstats,stats}" into ch_mlib_bam_stats_mqc + file "*.txt" into ch_mlib_bam_metrics_mqc script: - def prefix="${name}.mLb.mkD" - def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + prefix = "${name}.mLb.mkD" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() def avail_mem = 3 - if (!task.memory){ + if (!task.memory) { log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." } else { avail_mem = task.memory.toGiga() @@ -636,21 +679,21 @@ process MergeLibrary { if (bam_files.size() > 1) { """ picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp samtools index ${name}.sorted.bam picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp samtools index ${prefix}.sorted.bam samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats @@ -660,13 +703,13 @@ process MergeLibrary { } else { """ picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${bam_files[0]} \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=false \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + INPUT=${bam_files[0]} \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=false \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp samtools index ${prefix}.sorted.bam samtools idxstats ${prefix}.sorted.bam > ${prefix}.sorted.bam.idxstats @@ -679,7 +722,7 @@ process MergeLibrary { /* * STEP 4.2 Filter BAM file at merged library-level */ -process MergeLibraryFilter { +process MergeLibraryBAMFilter { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -694,31 +737,32 @@ process MergeLibraryFilter { } input: - set val(name), file(bam) from mlib_bam_filter - file bed from genome_filter_regions.collect() + set val(name), file(bam) from ch_mlib_bam_filter + file bed from ch_genome_filter_regions.collect() file bamtools_filter_config from ch_bamtools_filter_config output: - set val(name), file("*.{bam,bam.bai}") into mlib_filter_bam - set val(name), file("*.flagstat") into mlib_filter_flagstat - file "*.{idxstats,stats}" into mlib_filter_stats_mqc + set val(name), file("*.{bam,bam.bai}") into ch_mlib_filter_bam + set val(name), file("*.flagstat") into ch_mlib_filter_bam_flagstat + file "*.{idxstats,stats}" into ch_mlib_filter_bam_stats_mqc script: - def prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" - def filter_params = params.singleEnd ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" - def dup_params = params.keepDups ? "" : "-F 0x0400" - def multimap_params = params.keepMultiMap ? "" : "-q 1" - def name_sort_bam = params.singleEnd ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" + prefix = params.singleEnd ? "${name}.mLb.clN" : "${name}.mLb.flT" + filter_params = params.singleEnd ? "-F 0x004" : "-F 0x004 -F 0x0008 -f 0x001" + dup_params = params.keepDups ? "" : "-F 0x0400" + multimap_params = params.keepMultiMap ? "" : "-q 1" + blacklist_params = params.blacklist ? "-L $bed" : "" + name_sort_bam = params.singleEnd ? "" : "samtools sort -n -@ $task.cpus -o ${prefix}.bam -T $prefix ${prefix}.sorted.bam" """ samtools view \\ - $filter_params \\ - $dup_params \\ - $multimap_params \\ - -L $bed \\ - -b ${bam[0]} \\ - | bamtools filter \\ - -out ${prefix}.sorted.bam \\ - -script $bamtools_filter_config + $filter_params \\ + $dup_params \\ + $multimap_params \\ + $blacklist_params \\ + -b ${bam[0]} \\ + | bamtools filter \\ + -out ${prefix}.sorted.bam \\ + -script $bamtools_filter_config samtools index ${prefix}.sorted.bam samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat @@ -732,19 +776,20 @@ process MergeLibraryFilter { /* * STEP 4.3 Remove orphan reads from paired-end BAM file */ -if (params.singleEnd){ - mlib_filter_bam.into { mlib_rm_orphan_bam_metrics; - mlib_rm_orphan_bam_bigwig; - mlib_rm_orphan_bam_macs; - mlib_rm_orphan_bam_mrep; - mlib_name_bam_mlib_counts; - mlib_name_bam_mrep_counts } - mlib_filter_flagstat.into { mlib_rm_orphan_flagstat_bigwig; - mlib_rm_orphan_flagstat_macs; - mlib_rm_orphan_flagstat_mqc } - mlib_filter_stats_mqc.set { mlib_rm_orphan_stats_mqc } +if (params.singleEnd) { + ch_mlib_filter_bam.into { ch_mlib_rm_orphan_bam_metrics; + ch_mlib_rm_orphan_bam_bigwig; + ch_mlib_rm_orphan_bam_macs; + ch_mlib_rm_orphan_bam_plotfingerprint; + ch_mlib_rm_orphan_bam_mrep; + ch_mlib_name_bam_mlib_counts; + ch_mlib_name_bam_mrep_counts } + ch_mlib_filter_bam_flagstat.into { ch_mlib_rm_orphan_flagstat_bigwig; + ch_mlib_rm_orphan_flagstat_macs; + ch_mlib_rm_orphan_flagstat_mqc } + ch_mlib_filter_bam_stats_mqc.set { ch_mlib_rm_orphan_stats_mqc } } else { - process MergeLibraryRemoveOrphanReads { + process MergeLibraryBAMRemoveOrphan { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -758,22 +803,23 @@ if (params.singleEnd){ } input: - set val(name), file(bam) from mlib_filter_bam + set val(name), file(bam) from ch_mlib_filter_bam output: - set val(name), file("*.sorted.{bam,bam.bai}") into mlib_rm_orphan_bam_metrics, - mlib_rm_orphan_bam_bigwig, - mlib_rm_orphan_bam_macs, - mlib_rm_orphan_bam_mrep - set val(name), file("${prefix}.bam") into mlib_name_bam_mlib_counts, - mlib_name_bam_mrep_counts - set val(name), file("*.flagstat") into mlib_rm_orphan_flagstat_bigwig, - mlib_rm_orphan_flagstat_macs, - mlib_rm_orphan_flagstat_mqc - file "*.{idxstats,stats}" into mlib_rm_orphan_stats_mqc + set val(name), file("*.sorted.{bam,bam.bai}") into ch_mlib_rm_orphan_bam_metrics, + ch_mlib_rm_orphan_bam_bigwig, + ch_mlib_rm_orphan_bam_macs, + ch_mlib_rm_orphan_bam_plotfingerprint, + ch_mlib_rm_orphan_bam_mrep + set val(name), file("${prefix}.bam") into ch_mlib_name_bam_mlib_counts, + ch_mlib_name_bam_mrep_counts + set val(name), file("*.flagstat") into ch_mlib_rm_orphan_flagstat_bigwig, + ch_mlib_rm_orphan_flagstat_macs, + ch_mlib_rm_orphan_flagstat_mqc + file "*.{idxstats,stats}" into ch_mlib_rm_orphan_stats_mqc script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - def prefix="${name}.mLb.clN" + prefix = "${name}.mLb.clN" """ bampe_rm_orphan.py ${bam[0]} ${prefix}.bam --only_fr_pairs @@ -794,10 +840,34 @@ if (params.singleEnd){ /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// +// /* +// * STEP 5.1 preseq analysis after merging libraries and before filtering +// */ +// process MergeLibraryBAMPreseq { +// tag "$name" +// label 'process_low' +// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' +// +// when: +// !params.skipPreseq +// +// input: +// set val(name), file(bam) from ch_mlib_bam_preseq +// +// output: +// file "*.ccurve.txt" into ch_mlib_preseq_mqc +// +// script: +// prefix = "${name}.mLb.mkD" +// """ +// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} +// """ +// } + /* * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries */ -process MergeLibraryCollectMultipleMetrics { +process MergeLibraryBAMMetrics { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -807,29 +877,32 @@ process MergeLibraryCollectMultipleMetrics { else null } + when: + !params.skipPicardMetrics + input: - set val(name), file(bam) from mlib_rm_orphan_bam_metrics + set val(name), file(bam) from ch_mlib_rm_orphan_bam_metrics file fasta from ch_fasta output: - file "*_metrics" into mlib_collectmetrics_mqc - file "*.pdf" into mlib_collectmetrics_pdf + file "*_metrics" into ch_mlib_collectmetrics_mqc + file "*.pdf" into ch_mlib_collectmetrics_pdf script: - def prefix="${name}.mLb.clN" + prefix = "${name}.mLb.clN" def avail_mem = 3 - if (!task.memory){ + if (!task.memory) { log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." } else { avail_mem = task.memory.toGiga() } """ picard -Xmx${avail_mem}g CollectMultipleMetrics \\ - INPUT=${bam[0]} \\ - OUTPUT=${prefix}.CollectMultipleMetrics \\ - REFERENCE_SEQUENCE=$fasta \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + INPUT=${bam[0]} \\ + OUTPUT=${prefix}.CollectMultipleMetrics \\ + REFERENCE_SEQUENCE=$fasta \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp """ } @@ -841,66 +914,154 @@ process MergeLibraryBigWig { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', saveAs: {filename -> - if (filename.endsWith(".txt")) "scale/$filename" + if (filename.endsWith("scale_factor.txt")) "scale/$filename" else if (filename.endsWith(".bigWig")) "$filename" else null } input: - set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_bigwig.join(mlib_rm_orphan_flagstat_bigwig, by: [0]) - file sizes from genome_sizes_mlib_bigwig.collect() + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_bigwig.join(ch_mlib_rm_orphan_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mlib_bigwig.collect() output: - file "*.bigWig" into mlib_bigwig_igv - file "*.txt" into mlib_bigwig_scale + set val(name), file("*.bigWig") into ch_mlib_bigwig_plotprofile + file "*scale_factor.txt" into ch_mlib_bigwig_scale + file "*igv.txt" into ch_mlib_bigwig_igv script: - def prefix="${name}.mLb.clN" - def pe_fragment = params.singleEnd ? "" : "-pc" - def extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + prefix = "${name}.mLb.clN" + pe_fragment = params.singleEnd ? "" : "-pc" + extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -k1,1 -k2,2n > ${prefix}.bedGraph bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedLibrary/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt """ } +// /* +// * STEP 5.4 generate gene body coverage plot with deepTools +// */ +// process MergeLibraryPlotProfile { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' +// +// when: +// !params.skipPlotProfile +// +// input: +// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile +// file bed from ch_gene_bed +// +// output: +// file '*.{gz,pdf}' into ch_mlib_plotprofile_results +// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// """ +// computeMatrix scale-regions \\ +// --regionsFileName $bed \\ +// --scoreFileName $bigwig \\ +// --outFileName ${prefix}.computeMatrix.mat.gz \\ +// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ +// --regionBodyLength 1000 \\ +// --beforeRegionStartLength 3000 \\ +// --afterRegionStartLength 3000 \\ +// --skipZeros \\ +// --smartLabels \\ +// -p $task.cpus +// +// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ +// --outFileName ${prefix}.plotProfile.pdf \\ +// --outFileNameData ${prefix}.plotProfile.tab +// """ +// } + +// /* +// * STEP 6.1 deepTools plotFingerprint +// */ +// process MergeLibraryPlotFingerprint { +// tag "$name" +// label 'process_high' +// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' +// +// when: +// !params.skipPlotFingerprint +// +// input: +// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint +// +// output: +// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results +// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc +// +// script: +// prefix = "${name}.mLb.clN" +// extend = (params.singleEnd && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' +// """ +// plotFingerprint \\ +// --bamfiles ${bam[0]} \\ +// --plotFile ${prefix}.plotFingerprint.pdf \\ +// $extend \\ +// --labels $prefix \\ +// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ +// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ +// --skipZeros \\ +// --numberOfProcessors ${task.cpus} \\ +// --numberOfSamples ${params.fingerprint_bins} +// """ +// } + +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// +/* -- -- */ +/* -- MERGE LIBRARY PEAK ANALYSIS -- */ +/* -- -- */ +/////////////////////////////////////////////////////////////////////////////// +/////////////////////////////////////////////////////////////////////////////// + /* * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score */ process MergeLibraryMACSCallPeak { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}", mode: 'copy', saveAs: {filename -> if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null else filename } + when: + params.macs_gsize + input: - set val(name), file(bam), file(flagstat) from mlib_rm_orphan_bam_macs.join(mlib_rm_orphan_flagstat_macs, by: [0]) + set val(name), file(bam), file(flagstat) from ch_mlib_rm_orphan_bam_macs.join(ch_mlib_rm_orphan_flagstat_macs, by: [0]) file mlib_peak_count_header from ch_mlib_peak_count_header file mlib_frip_score_header from ch_mlib_frip_score_header output: - set val(name), file("*.{bed,xls,gappedPeak}") into mlib_macs_output - set val(name), file("*$peakext") into mlib_macs_peaks_homer, - mlib_macs_peaks_qc, - mlib_macs_peaks_consensus, - mlib_macs_peaks_ataqv, - mlib_macs_peaks_igv - file "*_mqc.tsv" into mlib_macs_peaks_mqc - - when: - params.macs_gsize + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mlib_macs_output + set val(name), file("*$peaktype") into ch_mlib_macs_homer, + ch_mlib_macs_qc, + ch_mlib_macs_consensus, + ch_mlib_macs_ataqv + file "*igv.txt" into ch_mlib_macs_igv + file "*_mqc.tsv" into ch_mlib_macs_mqc script: - def prefix="${name}.mLb.clN" - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - def broad = params.narrowPeak ? '' : "--broad" - def format = params.singleEnd ? "BAM" : "BAMPE" + prefix = "${name}.mLb.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + broad = params.narrowPeak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.singleEnd ? "BAM" : "BAMPE" + pileup = params.saveMACSPileup ? "-B --SPMR" : "" """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -908,13 +1069,16 @@ process MergeLibraryMACSCallPeak { -f $format \\ -g ${params.macs_gsize} \\ -n ${prefix} \\ + $pileup \\ --keep-dup all \\ --nomodel - cat ${prefix}_peaks${peakext} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + cat ${prefix}_peaks.${peaktype} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks${peakext} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${peaktype} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mlib_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${peaktype}" -exec echo -e "bwa/mergedLibrary/macs/${peaktype}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt """ } @@ -924,27 +1088,29 @@ process MergeLibraryMACSCallPeak { process MergeLibraryAnnotatePeaks { tag "$name" label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}", mode: 'copy' + + when: + params.macs_gsize input: - set val(name), file(peak) from mlib_macs_peaks_homer + set val(name), file(peak) from ch_mlib_macs_homer file fasta from ch_fasta file gtf from ch_gtf output: - file "*.txt" into mlib_macs_annotate - - when: - params.macs_gsize + file "*.txt" into ch_mlib_macs_annotate script: - def prefix="${name}.mLb.clN" + prefix = "${name}.mLb.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" """ - annotatePeaks.pl $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - > ${prefix}_peaks.annotatePeaks.txt + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + > ${prefix}_peaks.annotatePeaks.txt """ } @@ -952,74 +1118,82 @@ process MergeLibraryAnnotatePeaks { * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergeLibraryPeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/qc", mode: 'copy' - - input: - file peaks from mlib_macs_peaks_qc.collect{ it[1] } - file annos from mlib_macs_annotate.collect() - file mlib_peak_annotation_header from ch_mlib_peak_annotation_header - - output: - file "*.{txt,pdf}" into mlib_macs_qc - file "*.tsv" into mlib_macs_qc_mqc - - when: - params.macs_gsize - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - def suffix='mLb.clN' - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - """ - plot_macs_qc.r -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks${peakext}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ + label "process_medium" + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/qc", mode: 'copy' + + when: + params.macs_gsize + + input: + file peaks from ch_mlib_macs_qc.collect{ it[1] } + file annos from ch_mlib_macs_annotate.collect() + file mlib_peak_annotation_header from ch_mlib_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mlib_peak_qc + file "*.tsv" into ch_mlib_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mLb.clN' + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${peaktype}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mlib_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ } /* * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ process MergeLibraryCreateConsensusPeakSet { - publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' + label 'process_long' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus", mode: 'copy', + saveAs: {filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && (multipleGroups || replicatesExist) input: - file peaks from mlib_macs_peaks_consensus.collect{ it[1] } + file peaks from ch_mlib_macs_consensus.collect{ it[1] } output: - file "*.bed" into mlib_macs_consensus_bed, - mlib_macs_consensus_bed_igv - file "*.boolean.txt" into mlib_macs_consensus_bool - file "*.saf" into mlib_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into mlib_macs_consensus_intersect - - when: - params.macs_gsize && (multiple_samples || replicates_exist) + file "*.bed" into ch_mlib_macs_consensus_bed + file "*.saf" into ch_mlib_macs_consensus_saf + file "*.boolean.txt" into ch_mlib_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mlib_macs_consensus_intersect + file "*igv.txt" into ch_mlib_macs_consensus_igv script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - def suffix='mLb.clN' - def prefix="consensus_peaks.${suffix}" - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - def mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') - def collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - def expandparam = params.narrowPeak ? "--is_narrow_peak" : "" + suffix = 'mLb.clN' + prefix = "consensus_peaks.${suffix}" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - macs2_merged_expand.py ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks${peakext}","")} \\ - ${prefix}.boolean.txt \\ - --min_samples 1 \\ - $expandparam + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${peaktype}","")} \\ + ${prefix}.boolean.txt \\ + --min_replicates $params.min_reps_consensus \\ + $expandparam awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed @@ -1028,6 +1202,8 @@ process MergeLibraryCreateConsensusPeakSet { sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedLibrary/macs/${peaktype}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt """ } @@ -1036,28 +1212,30 @@ process MergeLibraryCreateConsensusPeakSet { */ process MergeLibraryAnnotateConsensusPeakSet { label "process_medium" - publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus", mode: 'copy' + + when: + params.macs_gsize && (multipleGroups || replicatesExist) input: - file bed from mlib_macs_consensus_bed - file bool from mlib_macs_consensus_bool + file bed from ch_mlib_macs_consensus_bed + file bool from ch_mlib_macs_consensus_bool file fasta from ch_fasta file gtf from ch_gtf output: - file "*.annotatePeaks.txt" into mlib_macs_consensus_annotate - - when: - params.macs_gsize && (multiple_samples || replicates_exist) + file "*.annotatePeaks.txt" into ch_mlib_macs_consensus_annotate script: - def prefix="consensus_peaks.mLb.clN" + prefix = "consensus_peaks.mLb.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" """ - annotatePeaks.pl $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - > ${prefix}.annotatePeaks.txt + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + > ${prefix}.annotatePeaks.txt cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt @@ -1069,45 +1247,54 @@ process MergeLibraryAnnotateConsensusPeakSet { */ process MergeLibraryDeseqConsensusPeakSet { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/macs/consensus/deseq2", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus/deseq2", mode: 'copy', + saveAs: {filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + params.macs_gsize && !params.skipDiffAnalysis && multipleGroups && replicatesExist input: - file bams from mlib_name_bam_mlib_counts.collect{ it[1] } - file saf from mlib_macs_consensus_saf.collect() + file bams from ch_mlib_name_bam_mlib_counts.collect{ it[1] } + file saf from ch_mlib_macs_consensus_saf.collect() file mlib_deseq2_pca_header from ch_mlib_deseq2_pca_header file mlib_deseq2_clustering_header from ch_mlib_deseq2_clustering_header output: - file "*featureCounts.txt" into mlib_macs_consensus_counts - file "*featureCounts.txt.summary" into mlib_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into mlib_macs_consensus_deseq_results - file "sizeFactors" into mlib_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into mlib_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into mlib_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into mlib_macs_consensus_deseq_mqc - - when: - params.macs_gsize && multiple_samples && replicates_exist + file "*featureCounts.txt" into ch_mlib_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mlib_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mlib_macs_consensus_deseq_results + file "sizeFactors" into ch_mlib_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mlib_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mlib_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mlib_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mlib_macs_consensus_deseq_mqc script: - def prefix="consensus_peaks.mLb.clN" - def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - def pe_params = params.singleEnd ? '' : "-p --donotsort" + prefix = "consensus_peaks.mLb.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + pe_params = params.singleEnd ? '' : "-p --donotsort" """ - featureCounts -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb cat $mlib_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mlib_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedLibrary/macs/${peaktype}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt """ } @@ -1115,55 +1302,57 @@ process MergeLibraryDeseqConsensusPeakSet { * STEP 4.6.1 Run ataqv on BAM file and corresponding peaks */ process MergeLibraryAtaqv { - tag "$name" - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' - - input: - set val(name), file(bam), file(peak) from mlib_bam_ataqv.join(mlib_macs_peaks_ataqv, by: [0]) - file tss_bed from ch_tss_bed - file autosomes from genome_autosomes.collect() - - output: - file "*.json" into mlib_ataqv - - script: - def peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" - def mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" - """ - ataqv --threads $task.cpus \\ - $peak_param \\ - --tss-file $tss_bed \\ - --metrics-file ${name}.ataqv.json \\ - --name $name \\ - --ignore-read-groups \\ - --autosomal-reference-file $autosomes \\ - $mito_param \\ - NA \\ - ${bam[0]} - """ + tag "$name" + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' + + input: + set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) + file autosomes from ch_genome_autosomes.collect() + file tss_bed from ch_tss_bed + + output: + file "*.json" into ch_mlib_ataqv + + script: + peak_param = params.macs_gsize ? "--peak-file ${peak}" : "" + mito_param = params.mito_name ? "--mitochondrial-reference-name ${params.mito_name}" : "" + """ + ataqv \\ + --threads $task.cpus \\ + $peak_param \\ + --tss-file $tss_bed \\ + --metrics-file ${name}.ataqv.json \\ + --name $name \\ + --ignore-read-groups \\ + --autosomal-reference-file $autosomes \\ + $mito_param \\ + NA \\ + ${bam[0]} + """ } /* * STEP 4.6.2 run ataqv mkarv on all JSON files to render web app */ process MergeLibraryAtaqvMkarv { - label 'process_medium' - publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' - - input: - file json from mlib_ataqv.collect() - - output: - file "html" into mlib_ataqv_mkarv - - script: - """ - mkarv --concurrency $task.cpus \\ - --force \\ - ./html/ \\ - ${json.join(' ')} - """ + label 'process_medium' + publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' + + input: + file json from ch_mlib_ataqv.collect() + + output: + file "html" into ch_mlib_ataqv_mkarv + + script: + """ + mkarv \\ + --concurrency $task.cpus \\ + --force \\ + ./html/ \\ + ${json.join(' ')} + """ } /////////////////////////////////////////////////////////////////////////////// @@ -1177,12 +1366,12 @@ process MergeLibraryAtaqvMkarv { /* * STEP 5.1 Merge library BAM files across all replicates */ -mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } - .groupTuple(by: [0]) - .map { it -> [ it[0], it[1].flatten() ] } - .set { mlib_rm_orphan_bam_mrep } +ch_mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } + .groupTuple(by: [0]) + .map { it -> [ it[0], it[1].flatten() ] } + .set { ch_mlib_rm_orphan_bam_mrep } -process MergeReplicate { +process MergeReplicateBAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', @@ -1195,25 +1384,25 @@ process MergeReplicate { } input: - set val(name), file(bams) from mlib_rm_orphan_bam_mrep + set val(name), file(bams) from ch_mlib_rm_orphan_bam_mrep output: - set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into mrep_bam_bigwig, - mrep_bam_macs - set val(name), file("*.flagstat") into mrep_flagstat_bigwig, - mrep_flagstat_macs, - mrep_flagstat_mqc - file "*.{idxstats,stats}" into mrep_stats_mqc - file "*.txt" into mrep_metrics_mqc + set val(name), file("*${prefix}.sorted.{bam,bam.bai}") into ch_mrep_bam_bigwig, + ch_mrep_bam_macs + set val(name), file("*.flagstat") into ch_mrep_bam_flagstat_bigwig, + ch_mrep_bam_flagstat_macs, + ch_mrep_bam_flagstat_mqc + file "*.{idxstats,stats}" into ch_mrep_bam_stats_mqc + file "*.txt" into ch_mrep_bam_metrics_mqc when: - !params.skipMergeReplicates && replicates_exist + !params.skipMergeReplicates && replicatesExist script: - def prefix="${name}.mRp.clN" - def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + prefix = "${name}.mRp.clN" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() def avail_mem = 3 - if (!task.memory){ + if (!task.memory) { log.info "[Picard MarkDuplicates] Available memory not known - defaulting to 3GB. Specify process memory requirements to change this." } else { avail_mem = task.memory.toGiga() @@ -1221,21 +1410,21 @@ process MergeReplicate { if (bam_files.size() > 1) { """ picard -Xmx${avail_mem}g MergeSamFiles \\ - ${'INPUT='+bam_files.join(' INPUT=')} \\ - OUTPUT=${name}.sorted.bam \\ - SORT_ORDER=coordinate \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + ${'INPUT='+bam_files.join(' INPUT=')} \\ + OUTPUT=${name}.sorted.bam \\ + SORT_ORDER=coordinate \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp samtools index ${name}.sorted.bam picard -Xmx${avail_mem}g MarkDuplicates \\ - INPUT=${name}.sorted.bam \\ - OUTPUT=${prefix}.sorted.bam \\ - ASSUME_SORTED=true \\ - REMOVE_DUPLICATES=true \\ - METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ - VALIDATION_STRINGENCY=LENIENT \\ - TMP_DIR=tmp + INPUT=${name}.sorted.bam \\ + OUTPUT=${prefix}.sorted.bam \\ + ASSUME_SORTED=true \\ + REMOVE_DUPLICATES=true \\ + METRICS_FILE=${prefix}.MarkDuplicates.metrics.txt \\ + VALIDATION_STRINGENCY=LENIENT \\ + TMP_DIR=tmp samtools index ${prefix}.sorted.bam samtools flagstat ${prefix}.sorted.bam > ${prefix}.sorted.bam.flagstat @@ -1263,75 +1452,80 @@ process MergeReplicate { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 5.2.1 Read depth normalised bigWig + * STEP 4.4.2 Read depth normalised bigWig */ process MergeReplicateBigWig { tag "$name" - label 'process_long' + label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', saveAs: {filename -> - if (filename.endsWith(".txt")) "scale/$filename" + if (filename.endsWith("scale_factor.txt")) "scale/$filename" else if (filename.endsWith(".bigWig")) "$filename" else null } + when: + !params.skipMergeReplicates && replicatesExist + input: - set val(name), file(bam), file(flagstat) from mrep_bam_bigwig.join(mrep_flagstat_bigwig, by: [0]) - file sizes from genome_sizes_mrep_bigwig.collect() + set val(name), file(bam), file(flagstat) from ch_mrep_bam_bigwig.join(ch_mrep_bam_flagstat_bigwig, by: [0]) + file sizes from ch_genome_sizes_mrep_bigwig.collect() output: - file "*.bigWig" into mrep_bigwig_igv - file "*.txt" into mrep_bigwig_scale - - when: - !params.skipMergeReplicates && replicates_exist + set val(name), file("*.bigWig") into ch_mrep_bigwig + file "*scale_factor.txt" into ch_mrep_bigwig_scale + file "*igv.txt" into ch_mrep_bigwig_igv script: - def prefix="${name}.mRp.clN" - def pe_fragment = params.singleEnd ? "" : "-pc" - def extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' + prefix = "${name}.mRp.clN" + pe_fragment = params.singleEnd ? "" : "-pc" + extend = (params.singleEnd && params.fragment_size > 0) ? "-fs ${params.fragment_size}" : '' """ SCALE_FACTOR=\$(grep 'mapped (' $flagstat | awk '{print 1000000/\$1}') echo \$SCALE_FACTOR > ${prefix}.scale_factor.txt genomeCoverageBed -ibam ${bam[0]} -bg -scale \$SCALE_FACTOR $pe_fragment $extend | sort -k1,1 -k2,2n > ${prefix}.bedGraph bedGraphToBigWig ${prefix}.bedGraph $sizes ${prefix}.bigWig + + find * -type f -name "*.bigWig" -exec echo -e "bwa/mergedReplicate/bigwig/"{}"\\t0,0,178" \\; > ${prefix}.bigWig.igv.txt """ } /* - * STEP 5.2.2 Call peaks with MACS2 and calculate FRiP score + * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score */ process MergeReplicateMACSCallPeak { tag "$name" label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy', + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}", mode: 'copy', saveAs: {filename -> if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null else filename } + when: + !params.skipMergeReplicates && replicatesExist && params.macs_gsize + input: - set val(name), file(bam), file(flagstat) from mrep_bam_macs.join(mrep_flagstat_macs, by: [0]) + set val(name), file(bam), file(flagstat) from ch_mrep_bam_macs.join(ch_mrep_bam_flagstat_macs, by: [0]) file mrep_peak_count_header from ch_mrep_peak_count_header file mrep_frip_score_header from ch_mrep_frip_score_header output: - file "*.{bed,xls,gappedPeak}" into mrep_macs_output - set val(name), file("*$peakext") into mrep_macs_peaks_homer, - mrep_macs_peaks_qc, - mrep_macs_peaks_consensus, - mrep_macs_peaks_igv - file "*_mqc.tsv" into mrep_macs_peak_mqc - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize + set val(name), file("*.{bed,xls,gappedPeak,bdg}") into ch_mrep_macs_output + set val(name), file("*$peaktype") into ch_mrep_macs_homer, + ch_mrep_macs_qc, + ch_mrep_macs_consensus + file "*igv.txt" into ch_mrep_macs_igv + file "*_mqc.tsv" into ch_mrep_macs_mqc script: - def prefix="${name}.mRp.clN" - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - def broad = params.narrowPeak ? '' : "--broad" - def format = params.singleEnd ? "BAM" : "BAMPE" + prefix = "${name}.mRp.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + broad = params.narrowPeak ? '' : "--broad --broad-cutoff ${params.broad_cutoff}" + format = params.singleEnd ? "BAM" : "BAMPE" + pileup = params.saveMACSPileup ? "-B --SPMR" : "" """ macs2 callpeak \\ -t ${bam[0]} \\ @@ -1339,118 +1533,130 @@ process MergeReplicateMACSCallPeak { -f $format \\ -g ${params.macs_gsize} \\ -n ${prefix} \\ + $pileup \\ --keep-dup all \\ --nomodel - cat ${prefix}_peaks${peakext} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv + cat ${prefix}_peaks.${peaktype} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv - READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks${peakext} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') + READS_IN_PEAKS=\$(intersectBed -a ${bam[0]} -b ${prefix}_peaks.${peaktype} -bed -c -f 0.20 | awk -F '\t' '{sum += \$NF} END {print sum}') grep 'mapped (' $flagstat | awk -v a="\$READS_IN_PEAKS" -v OFS='\t' '{print "${name}", a/\$1}' | cat $mrep_frip_score_header - > ${prefix}_peaks.FRiP_mqc.tsv + + find * -type f -name "*.${peaktype}" -exec echo -e "bwa/mergedReplicate/macs/${peaktype}/"{}"\\t0,0,178" \\; > ${prefix}_peaks.igv.txt """ } /* - * STEP 5.2.3 Annotate peaks with HOMER + * STEP 4.5.2 Annotate peaks with HOMER */ process MergeReplicateAnnotatePeaks { tag "$name" label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}", mode: 'copy' + + when: + !params.skipMergeReplicates && replicatesExist && params.macs_gsize input: - set val(name), file(peak) from mrep_macs_peaks_homer + set val(name), file(peak) from ch_mrep_macs_homer file fasta from ch_fasta file gtf from ch_gtf output: - file "*.txt" into mrep_macs_annotate - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize + file "*.txt" into ch_mrep_macs_annotate script: - def prefix="${name}.mRp.clN" + prefix = "${name}.mRp.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" """ - annotatePeaks.pl $peak \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - > ${prefix}_peaks.annotatePeaks.txt + annotatePeaks.pl \\ + $peak \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + > ${prefix}_peaks.annotatePeaks.txt """ } /* - * STEP 5.2.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergeReplicatePeakQC { - label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/qc", mode: 'copy' - - input: - file peaks from mrep_macs_peaks_qc.collect{ it[1] } - file annos from mrep_macs_annotate.collect() - file mrep_peak_annotation_header from ch_mrep_peak_annotation_header - - output: - file "*.{txt,pdf}" into mrep_macs_qc - file "*.tsv" into mrep_macs_qc_mqc - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize - - script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ - def suffix='mRp.clN' - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - """ - plot_macs_qc.r -i ${peaks.join(',')} \\ - -s ${peaks.join(',').replaceAll(".${suffix}_peaks${peakext}","")} \\ - -o ./ \\ - -p macs_peak.${suffix} - - plot_homer_annotatepeaks.r -i ${annos.join(',')} \\ - -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ - -o ./ \\ - -p macs_annotatePeaks.${suffix} - - cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv - """ + label "process_medium" + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/qc", mode: 'copy' + + when: + !params.skipMergeReplicates && replicatesExist && params.macs_gsize + + input: + file peaks from ch_mrep_macs_qc.collect{ it[1] } + file annos from ch_mrep_macs_annotate.collect() + file mrep_peak_annotation_header from ch_mrep_peak_annotation_header + + output: + file "*.{txt,pdf}" into ch_mrep_peak_qc + file "*.tsv" into ch_mrep_peak_qc_mqc + + script: // This script is bundled with the pipeline, in nf-core/atacseq/bin/ + suffix = 'mRp.clN' + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + """ + plot_macs_qc.r \\ + -i ${peaks.join(',')} \\ + -s ${peaks.join(',').replaceAll(".${suffix}_peaks.${peaktype}","")} \\ + -o ./ \\ + -p macs_peak.${suffix} + + plot_homer_annotatepeaks.r \\ + -i ${annos.join(',')} \\ + -s ${annos.join(',').replaceAll(".${suffix}_peaks.annotatePeaks.txt","")} \\ + -o ./ \\ + -p macs_annotatePeaks.${suffix} + + cat $mrep_peak_annotation_header macs_annotatePeaks.${suffix}.summary.txt > macs_annotatePeaks.${suffix}.summary_mqc.tsv + """ } /* - * STEP 5.2.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ process MergeReplicateCreateConsensusPeakSet { - publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' + label 'process_long' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus", mode: 'copy', + saveAs: {filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skipMergeReplicates && replicatesExist && params.macs_gsize && multipleGroups input: - file peaks from mrep_macs_peaks_consensus.collect{ it[1] } + file peaks from ch_mrep_macs_consensus.collect{ it[1] } output: - file "*.bed" into mrep_macs_consensus_bed, - mrep_macs_consensus_bed_igv - file "*.boolean.txt" into mrep_macs_consensus_bool - file "*.saf" into mrep_macs_consensus_saf - file "*.intersect.{txt,plot.pdf}" into mrep_macs_consensus_intersect - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + file "*.bed" into ch_mrep_macs_consensus_bed + file "*.saf" into ch_mrep_macs_consensus_saf + file "*.boolean.txt" into ch_mrep_macs_consensus_bool + file "*.intersect.{txt,plot.pdf}" into ch_mrep_macs_consensus_intersect + file "*igv.txt" into ch_mrep_macs_consensus_igv script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - def suffix='mRp.clN' - def prefix="consensus_peaks.${suffix}" - def peakext = params.narrowPeak ? ".narrowPeak" : ".broadPeak" - def mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') - def collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') - def expandparam = params.narrowPeak ? "--is_narrow_peak" : "" + suffix = 'mRp.clN' + prefix = "consensus_peaks.${suffix}" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + mergecols = params.narrowPeak ? (2..10).join(',') : (2..9).join(',') + collapsecols = params.narrowPeak ? (["collapse"]*9).join(',') : (["collapse"]*8).join(',') + expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt - macs2_merged_expand.py ${prefix}.txt \\ - ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks${peakext}","")} \\ - ${prefix}.boolean.txt \\ - --min_samples 1 \\ - $expandparam + macs2_merged_expand.py \\ + ${prefix}.txt \\ + ${peaks.collect{it.toString()}.sort().join(',').replaceAll("_peaks.${peaktype}","")} \\ + ${prefix}.boolean.txt \\ + $expandparam awk -v FS='\t' -v OFS='\t' 'FNR > 1 { print \$1, \$2, \$3, \$4, "0", "+" }' ${prefix}.boolean.txt > ${prefix}.bed @@ -1459,36 +1665,40 @@ process MergeReplicateCreateConsensusPeakSet { sed -i 's/.${suffix}//g' ${prefix}.boolean.intersect.txt plot_peak_intersect.r -i ${prefix}.boolean.intersect.txt -o ${prefix}.boolean.intersect.plot.pdf + + find * -type f -name "${prefix}.bed" -exec echo -e "bwa/mergedReplicate/macs/${peaktype}/consensus/"{}"\\t0,0,0" \\; > ${prefix}.bed.igv.txt """ } /* - * STEP 5.2.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file + * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MergeReplicateAnnotateConsensusPeakSet { label "process_medium" - publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus", mode: 'copy' + + when: + !params.skipMergeReplicates && replicatesExist && params.macs_gsize && multipleGroups input: - file bed from mrep_macs_consensus_bed - file bool from mrep_macs_consensus_bool + file bed from ch_mrep_macs_consensus_bed + file bool from ch_mrep_macs_consensus_bool file fasta from ch_fasta file gtf from ch_gtf output: - file "*.annotatePeaks.txt" into mrep_macs_consensus_annotate - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + file "*.annotatePeaks.txt" into ch_mrep_macs_consensus_annotate script: - def prefix="consensus_peaks.mRp.clN" + prefix = "consensus_peaks.mRp.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" """ - annotatePeaks.pl $bed \\ - $fasta \\ - -gid \\ - -gtf $gtf \\ - > ${prefix}.annotatePeaks.txt + annotatePeaks.pl \\ + $bed \\ + $fasta \\ + -gid \\ + -gtf $gtf \\ + > ${prefix}.annotatePeaks.txt cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt paste $bool tmp.txt > ${prefix}.boolean.annotatePeaks.txt @@ -1496,49 +1706,58 @@ process MergeReplicateAnnotateConsensusPeakSet { } /* - * STEP 5.2.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ process MergeReplicateDeseqConsensusPeakSet { label 'process_medium' - publishDir "${params.outdir}/bwa/mergedReplicate/macs/consensus/deseq2", mode: 'copy' + publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus/deseq2", mode: 'copy', + saveAs: {filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } + + when: + !params.skipMergeReplicates && !params.skipDiffAnalysis && replicatesExist && params.macs_gsize && multipleGroups input: - file bams from mlib_name_bam_mrep_counts.collect{ it[1] } - file saf from mrep_macs_consensus_saf.collect() + file bams from ch_mlib_name_bam_mrep_counts.collect{ it[1] } + file saf from ch_mrep_macs_consensus_saf.collect() file mrep_deseq2_pca_header from ch_mrep_deseq2_pca_header file mrep_deseq2_clustering_header from ch_mrep_deseq2_clustering_header output: - file "*featureCounts.txt" into mrep_macs_consensus_counts - file "*featureCounts.txt.summary" into mrep_macs_consensus_counts_mqc - file "*.{RData,results.txt,pdf,log}" into mrep_macs_consensus_deseq_results - file "sizeFactors" into mrep_macs_consensus_deseq_factors - file "*vs*/*.{pdf,txt}" into mrep_macs_consensus_deseq_comp_results - file "*vs*/*.bed" into mrep_macs_consensus_deseq_comp_bed_igv - file "*.tsv" into mrep_macs_consensus_deseq_mqc - - when: - !params.skipMergeReplicates && replicates_exist && params.macs_gsize && multiple_samples + file "*featureCounts.txt" into ch_mrep_macs_consensus_counts + file "*featureCounts.txt.summary" into ch_mrep_macs_consensus_counts_mqc + file "*.{RData,results.txt,pdf,log}" into ch_mrep_macs_consensus_deseq_results + file "sizeFactors" into ch_mrep_macs_consensus_deseq_factors + file "*vs*/*.{pdf,txt}" into ch_mrep_macs_consensus_deseq_comp_results + file "*vs*/*.bed" into ch_mrep_macs_consensus_deseq_comp_bed + file "*igv.txt" into ch_mrep_macs_consensus_deseq_comp_igv + file "*.tsv" into ch_mrep_macs_consensus_deseq_mqc script: - def prefix="consensus_peaks.mRp.clN" - def bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() - def bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" - def pe_params = params.singleEnd ? '' : "-p --donotsort" + prefix = "consensus_peaks.mRp.clN" + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" + bam_files = bams.findAll { it.toString().endsWith('.bam') }.sort() + bam_ext = params.singleEnd ? ".mLb.clN.sorted.bam" : ".mLb.clN.bam" + pe_params = params.singleEnd ? '' : "-p --donotsort" """ - featureCounts -F SAF \\ - -O \\ - --fracOverlap 0.2 \\ - -T $task.cpus \\ - $pe_params \\ - -a $saf \\ - -o ${prefix}.featureCounts.txt \\ - ${bam_files.join(' ')} + featureCounts \\ + -F SAF \\ + -O \\ + --fracOverlap 0.2 \\ + -T $task.cpus \\ + $pe_params \\ + -a $saf \\ + -o ${prefix}.featureCounts.txt \\ + ${bam_files.join(' ')} - featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mRp + featurecounts_deseq2.r -i ${prefix}.featureCounts.txt -b '$bam_ext' -o ./ -p $prefix -s .mLb cat $mrep_deseq2_pca_header ${prefix}.pca.vals.txt > ${prefix}.pca.vals_mqc.tsv cat $mrep_deseq2_clustering_header ${prefix}.sample.dists.txt > ${prefix}.sample.dists_mqc.tsv + + find * -type f -name "*.FDR0.05.results.bed" -exec echo -e "bwa/mergedReplicate/macs/${peaktype}/consensus/deseq2/"{}"\\t255,0,0" \\; > ${prefix}.igv.txt """ } @@ -1554,30 +1773,32 @@ process MergeReplicateDeseqConsensusPeakSet { * STEP 6 - Create IGV session file */ process IGV { - publishDir "${params.outdir}/igv", mode: 'copy' + publishDir "${params.outdir}/igv/${peaktype}", mode: 'copy' + + when: + !params.skipIGV input: file fasta from ch_fasta - file ('bwa/mergedLibrary/bigwig/*') from mlib_bigwig_igv.collect() - file ('bwa/mergedLibrary/macs/*') from mlib_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/*') from mlib_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedLibrary/macs/consensus/deseq2/*') from mlib_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file bigwigs from ch_mlib_bigwig_igv.collect().ifEmpty([]) + file peaks from ch_mlib_macs_igv.collect().ifEmpty([]) + file consensus_peaks from ch_mlib_macs_consensus_igv.collect().ifEmpty([]) + file differential_peaks from ch_mlib_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/bigwig/*') from mrep_bigwig_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/*') from mrep_macs_peaks_igv.collect{it[1]}.ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/*') from mrep_macs_consensus_bed_igv.collect().ifEmpty([]) - file ('bwa/mergedReplicate/macs/consensus/deseq2/*') from mrep_macs_consensus_deseq_comp_bed_igv.collect().ifEmpty([]) + file rbigwigs from ch_mrep_bigwig_igv.collect().ifEmpty([]) + file rpeaks from ch_mrep_macs_igv.collect().ifEmpty([]) + file rconsensus_peaks from ch_mrep_macs_consensus_igv.collect().ifEmpty([]) + file rdifferential_peaks from ch_mrep_macs_consensus_deseq_comp_igv.collect().ifEmpty([]) output: - file "*.{txt,xml}" into igv_session + file "*.{txt,xml}" into ch_igv_session script: // scripts are bundled with the pipeline, in nf-core/atacseq/bin/ - def outdir_abspath = new File(params.outdir).getCanonicalPath().toString() + peaktype = params.narrowPeak ? "narrowPeak" : "broadPeak" """ - igv_get_files.sh ./ mLb $outdir_abspath > igv_files.txt - igv_get_files.sh ./ mRp $outdir_abspath >> igv_files.txt - igv_files_to_session.py igv_session.xml igv_files.txt ${outdir_abspath}/reference_genome/${fasta.getName()} + cat *.txt > igv_files.txt + igv_files_to_session.py igv_session.xml igv_files.txt ../../reference_genome/${fasta.getName()} --path_prefix '../../' """ } @@ -1613,6 +1834,7 @@ process get_software_versions { samtools --version > v_samtools.txt bedtools --version > v_bedtools.txt echo \$(bamtools --version 2>&1) > v_bamtools.txt + #echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true picard MarkDuplicates --version &> v_picard.txt || true echo \$(R --version 2>&1) > v_R.txt python -c "import pysam; print(pysam.__version__)" > v_pysam.txt @@ -1620,6 +1842,7 @@ process get_software_versions { touch v_homer.txt echo \$(ataqv --version 2>&1) > v_ataqv.txt echo \$(featureCounts -v 2>&1) > v_featurecounts.txt + #preseq &> v_preseq.txt multiqc --version > v_multiqc.txt scrape_software_versions.py &> software_versions_mqc.yaml """ @@ -1647,49 +1870,58 @@ ${summary.collect { k,v -> "
$k
${v ?: ' hnames.each { hname -> - if(hostname.contains(hname) && !workflow.profile.contains(prof)){ + if (hostname.contains(hname) && !workflow.profile.contains(prof)) { log.error "====================================================\n" + " ${c_red}WARNING!${c_reset} You are running with `-profile $workflow.profile`\n" + " but your machine hostname is ${c_white}'$hostname'${c_reset}\n" + From 8c8e26eb5a8f348b35be3f9b02b8254e7267d6dc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 18:25:27 +0100 Subject: [PATCH 044/149] Woops...accidental delete --- bin/igv_files_to_session.py | 121 ++++++++++++++++++++++++++++++++++++ 1 file changed, 121 insertions(+) create mode 100755 bin/igv_files_to_session.py diff --git a/bin/igv_files_to_session.py b/bin/igv_files_to_session.py new file mode 100755 index 00000000..6dc6aabe --- /dev/null +++ b/bin/igv_files_to_session.py @@ -0,0 +1,121 @@ +#!/usr/bin/env python + +####################################################################### +####################################################################### +## Created on July 4th 2018 to create IGV session file from file list +####################################################################### +####################################################################### + +import os +import argparse + +############################################ +############################################ +## PARSE ARGUMENTS +############################################ +############################################ + +Description = 'Create IGV session file from a list of files and associated colours - ".bed", ".bw", ".bigwig", ".tdf", ".gtf" files currently supported.' +Epilog = """Example usage: python igv_files_to_session.py """ + +argParser = argparse.ArgumentParser(description=Description, epilog=Epilog) + +## REQUIRED PARAMETERS +argParser.add_argument('XML_OUT', help="XML output file.") +argParser.add_argument('LIST_FILE', help="Tab-delimited file containing two columns i.e. file_name\tcolour. Header isnt required.") +argParser.add_argument('GENOME', help="Full path to genome fasta file or shorthand for genome available in IGV e.g. hg19.") + +## OPTIONAL PARAMETERS +argParser.add_argument('-pp', '--path_prefix', type=str, dest="PATH_PREFIX", default='', help="Path prefix to be added at beginning of all files in input list file.") +args = argParser.parse_args() + +############################################ +############################################ +## HELPER FUNCTIONS +############################################ +############################################ + +def makedir(path): + + if not len(path) == 0: + try: + os.makedirs(path) + except OSError as exception: + if exception.errno != errno.EEXIST: + raise + +############################################ +############################################ +## MAIN FUNCTION +############################################ +############################################ + +def igv_files_to_session(XMLOut,ListFile,Genome,PathPrefix=''): + + makedir(os.path.dirname(XMLOut)) + + fileList = [] + fin = open(ListFile,'r') + while True: + line = fin.readline() + if line: + ifile,colour = line.strip().split('\t') + if len(colour.strip()) == 0: + colour = '0,0,178' + fileList.append((PathPrefix.strip()+ifile,colour)) + else: + break + fout.close() + + ## ADD RESOURCES SECTION + XMLStr = '\n' + XMLStr += '\n' % (Genome) + XMLStr += '\t\n' + for ifile,colour in fileList: + XMLStr += '\t\t\n' % (ifile) + XMLStr += '\t\n' + + ## ADD PANEL SECTION + XMLStr += '\t\n' + for ifile,colour in fileList: + extension = os.path.splitext(ifile)[1].lower() + if extension in ['.bed','.broadpeak','.narrowpeak']: + XMLStr += '\t\t Date: Thu, 3 Oct 2019 18:37:10 +0100 Subject: [PATCH 045/149] Add preseq and deeptools --- assets/multiqc/multiqc_config.yaml | 13 +++++++++++-- 1 file changed, 11 insertions(+), 2 deletions(-) diff --git a/assets/multiqc/multiqc_config.yaml b/assets/multiqc/multiqc_config.yaml index d2f44b25..e76c08b7 100755 --- a/assets/multiqc/multiqc_config.yaml +++ b/assets/multiqc/multiqc_config.yaml @@ -3,8 +3,6 @@ report_comment: > analysis pipeline. For information about how to interpret these results, please see the documentation. -skip_generalstats: true - export_plots: true fn_clean_exts: @@ -17,6 +15,7 @@ fn_clean_exts: - 'clN' - 'mLb' - 'mRp' + - '_peaks' module_order: - fastqc: @@ -48,6 +47,11 @@ module_order: info: 'This section of the report shows SAMTools results after merging libraries and before filtering.' path_filters: - '*mLb.mkD.sorted.bam*' + - preseq: + name: 'Preseq (merged library; unfiltered)' + info: 'This section of the report shows Preseq results after merging libraries and before filtering.' + path_filters: + - '*mLb*' - samtools: name: 'SAMTools (merged library; filtered)' info: 'This section of the report shows SAMTools results after merging libraries and after filtering.' @@ -58,6 +62,11 @@ module_order: info: 'This section of the report shows picard results after merging libraries and after filtering.' path_filters: - '*mLb*' + - deeptools: + name: 'deepTools' + info: 'This section of the report shows QC plots generated by deepTools.' + path_filters: + - '*.plot*' - samtools: name: 'SAMTools (merged replicate; filtered)' info: 'This section of the report shows SAMTools results after merging replicates and before filtering.' From 8eaaad26bc47da90546429e0692093d8a68ea00d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 18:51:58 +0100 Subject: [PATCH 046/149] Update for PR --- CHANGELOG.md | 22 ++++++++++++++++++++-- 1 file changed, 20 insertions(+), 2 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index b8c0177a..87c68903 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -10,16 +10,34 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` * Merged in TEMPLATE branch for automated syncing +* Renamed all channels to start with `ch_` prefix +* Capitalised process names * Add quick start information to main README +* Add parameters: + * `--seq_center` + * `--fingerprint_bins` + * `--broad_cutoff` + * `--min_reps_consensus` + * `--saveMACSPileup` + * `--skipDiffAnalysis` + * `--skip*` for skipping QC steps +* [#35](https://github.com/nf-core/atacseq/issues/35) - Add deepTools plotFingerprint * [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config -* [#46](https://github.com/nf-core/atacseq/issues/46) - Bump resources for MAC2 and annotatePeaks.pl * [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly ### `Fixed` +* Smaller logo for completion email +* IGV uses relative instead of absolute paths * [#41](https://github.com/nf-core/atacseq/issues/41) - Docs: Add example plot images * [#44](https://github.com/nf-core/atacseq/issues/44) - Output directory missing: macs2/consensus/deseq2 -* Smaller logo for completion email + +### `Dependencies` + +* Added preseq v2.0.3 +* Added deeptools v3.2.1 +* Added r-xfun v0.3 +* Added gawk v4.2.1 ## [1.0.0] - 2019-04-09 From 73963398fea09c5de61874432e85b696b0bc5e5e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 3 Oct 2019 19:14:26 +0100 Subject: [PATCH 047/149] Fix markdownlint --- CHANGELOG.md | 24 ++++++++++++------------ 1 file changed, 12 insertions(+), 12 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 87c68903..7cf59f03 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -9,28 +9,28 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. ### `Added` +* [#35](https://github.com/nf-core/atacseq/issues/35) - Add deepTools plotFingerprint +* [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config +* [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly * Merged in TEMPLATE branch for automated syncing * Renamed all channels to start with `ch_` prefix * Capitalised process names * Add quick start information to main README * Add parameters: - * `--seq_center` - * `--fingerprint_bins` - * `--broad_cutoff` - * `--min_reps_consensus` - * `--saveMACSPileup` - * `--skipDiffAnalysis` - * `--skip*` for skipping QC steps -* [#35](https://github.com/nf-core/atacseq/issues/35) - Add deepTools plotFingerprint -* [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config -* [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly + * `--seq_center` + * `--fingerprint_bins` + * `--broad_cutoff` + * `--min_reps_consensus` + * `--saveMACSPileup` + * `--skipDiffAnalysis` + * `--skip*` for skipping QC steps ### `Fixed` -* Smaller logo for completion email -* IGV uses relative instead of absolute paths * [#41](https://github.com/nf-core/atacseq/issues/41) - Docs: Add example plot images * [#44](https://github.com/nf-core/atacseq/issues/44) - Output directory missing: macs2/consensus/deseq2 +* Smaller logo for completion email +* IGV uses relative instead of absolute paths ### `Dependencies` From 3dbd837c8aba2916645bcbfefd453cbd06797b78 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 4 Oct 2019 17:47:08 +0100 Subject: [PATCH 048/149] Update parameters --- docs/usage.md | 95 +++++++++++++++++++++++++++++++++++++++++---------- 1 file changed, 77 insertions(+), 18 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 0fa49919..212f5ef5 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -2,6 +2,9 @@ ## Table of contents + + +* [Table of contents](#table-of-contents) * [Introduction](#introduction) * [Running the pipeline](#running-the-pipeline) * [Updating the pipeline](#updating-the-pipeline) @@ -11,8 +14,9 @@ * [`--design`](#--design) * [Generic arguments](#generic-arguments) * [`--singleEnd`](#--singleend) - * [`--narrowPeak`](#--narrowpeak) + * [`--seq_center`](#--seq_center) * [`--fragment_size`](#--fragment_size) + * [`--fingerprint_bins`](#--fingerprint_bins) * [Reference genomes](#reference-genomes) * [`--genome` (using iGenomes)](#--genome-using-igenomes) * [`--fasta`](#--fasta) @@ -20,9 +24,9 @@ * [`--bwa_index`](#--bwa_index) * [`--gene_bed`](#--gene_bed) * [`--tss_bed`](#--tss_bed) - * [`--mito_name`](#--mito_name) * [`--macs_gsize`](#--macs_gsize) * [`--blacklist`](#--blacklist) + * [`--mito_name`](#--mito_name) * [`--saveGenomeIndex`](#--savegenomeindex) * [`--igenomesIgnore`](#--igenomesignore) * [Adapter trimming](#adapter-trimming) @@ -34,6 +38,13 @@ * [`--keepMultiMap`](#--keepmultimap) * [`--skipMergeReplicates`](#--skipmergereplicates) * [`--saveAlignedIntermediates`](#--savealignedintermediates) +* [Peaks](#peaks) + * [`--narrowPeak`](#--narrowpeak) + * [`--broad_cutoff`](#--broad_cutoff) + * [`--min_reps_consensus`](#--min_reps_consensus) + * [`--saveMACSPileup`](#--savemacspileup) + * [`--skipDiffAnalysis`](#--skipdiffanalysis) +* [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) * [Custom resource requests](#custom-resource-requests) @@ -54,6 +65,8 @@ * [`--plaintext_email`](#--plaintext_email) * [`--monochrome_logs`](#--monochrome_logs) * [`--multiqc_config`](#--multiqc_config) + + ## Introduction Nextflow handles job submissions on SLURM or other environments, and supervises running the jobs. Thus the Nextflow process must run until the pipeline is finished. We recommend that you put the process running in the background through `screen` / `tmux` or similar tool. Alternatively you can run nextflow within a cluster job submitted your job scheduler. @@ -96,15 +109,16 @@ First, go to the [nf-core/atacseq releases page](https://github.com/nf-core/atac This version number will be logged in reports when you run the pipeline, so that you'll know what you used when you look back in the future. + ## Main arguments ### `-profile` -Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! +Use this parameter to choose a configuration profile. Profiles can give configuration pre-sets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! If `-profile` is not specified at all the pipeline will be run locally and expects all software to be installed and available on the `PATH`. * `awsbatch` - * A generic configuration profile to be used with AWS Batch + * A generic configuration profile to be used with AWS Batch. * `conda` * A generic configuration profile to be used with [conda](https://conda.io/docs/) * Pulls most software from [Bioconda](https://bioconda.github.io/) @@ -113,7 +127,7 @@ If `-profile` is not specified at all the pipeline will be run locally and expec * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) * `singularity` * A generic configuration profile to be used with [Singularity](http://singularity.lbl.gov/) - * Pulls software from dockerhub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) + * Pulls software from DockerHub: [`nfcore/atacseq`](http://hub.docker.com/r/nfcore/atacseq/) * `test` * A profile with a complete configuration for automated testing * Includes links to test data so needs no other parameters @@ -179,11 +193,18 @@ By default, the pipeline expects paired-end data. If you have single-end data, s It is not possible to run a mixture of single-end and paired-end files in one run. -### `--narrowPeak` -MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode. +### `--seq_center` +Sequencing center information that will be added to read groups in BAM files. ### `--fragment_size` -Number of base pairs to extend single-end reads when creating bigWig files. Default: `0` +Number of base pairs to extend single-end reads when creating bigWig files. + +Default: `0` + +### `--fingerprint_bins` +Number of genomic bins to use when generating the deepTools fingerprint plot. Larger numbers will give a smoother profile, but take longer to run. + +Default: `500000` ## Reference genomes @@ -201,7 +222,7 @@ You can find the keys to specify the genomes in the [iGenomes config file](../co * _Drosophila_ * `--genome BDGP6` * _S. cerevisiae_ - * `--genome R64-1-1` + * `--genome 'R64-1-1'` > There are numerous others - check the config file for more. @@ -242,38 +263,38 @@ Full path to an existing BWA index for your reference genome including the base ``` ### `--gene_bed` -The full path to BED file for genome-wide gene intervals. This will be created from the GTF file if it isnt specified. +The full path to BED file for genome-wide gene intervals. This will be created from the GTF file if not specified. ```bash --gene_bed '[path to gene BED file]' ``` ### `--tss_bed` -The full path to BED file for genome-wide transcription start sites. This will be created from the gene BED file if it isnt specified. +The full path to BED file for genome-wide transcription start sites. This will be created from the gene BED file if not specified. ```bash --tss_bed '[path to tss BED file]' ``` ### `--macs_gsize` -[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. These have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter isnt specified then the MACS2 peak-calling and differential analysis will be skipped. +[Effective genome size](https://github.com/taoliu/MACS#-g--gsize) parameter required by MACS2. These have been provided when `--genome` is set as *GRCh37*, *GRCh38*, *GRCm38*, *WBcel235*, *BDGP6*, *R64-1-1*, *EF2*, *hg38*, *hg19* and *mm10*. For other genomes, if this parameter is not specified then the MACS2 peak-calling and differential analysis will be skipped. ```bash --macs_gsize 2.7e9 ``` -### `--mito_name` -Name of mitochondrial chomosome in reference assembly. Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config). +### `--blacklist` +If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter. ```bash ---mito_name chrM +--blacklist '[path to blacklisted regions]' ``` -### `--blacklist` -If provided, alignments that overlap with the regions in this file will be filtered out (see [ENCODE blacklists](https://sites.google.com/site/anshulkundaje/projects/blacklists)). The file should be in BED format. Blacklisted regions for *GRCh37*, *GRCh38*, *GRCm38*, *hg19*, *hg38*, *mm10* are bundled with the pipeline in the [`blacklists`](../assets/blacklists/) directory, and as such will be automatically used if any of those genomes are specified with the `--genome` parameter. +### `--mito_name` +Name of mitochondrial chomosome in reference assembly. Reads aligning to this contig are filtered out if a valid identifier is provided otherwise this step is skipped. Where possible these have been provided in the [`igenomes.config`](../conf/igenomes.config). ```bash ---blacklist '[path to blacklisted regions]' +--mito_name chrM ``` ### `--saveGenomeIndex` @@ -318,6 +339,44 @@ An additional series of steps are performed by the pipeline by merging the repli ### `--saveAlignedIntermediates` By default, intermediate BAM files will not be saved. The final BAM files created after the appropriate filtering step are always saved to limit storage usage. Set to true to also save other intermediate BAM files. +## Peaks + +### `--narrowPeak` +MACS2 is run by default with the [`--broad`](https://github.com/taoliu/MACS#--broad) flag. Specify this flag to call peaks in narrowPeak mode. + +### `--broad_cutoff` +Specifies broad cut-off value for MACS2. Only used when `--narrowPeak` isnt specified. Default: 0.1 + +### `--min_reps_consensus` +Number of biological replicates required from a given condition for a peak to contribute to a consensus peak . If you are confident you have good reproducibility amongst your replicates then you can increase the value of this parameter to create a "reproducible" set of consensus of peaks. For example, a value of 2 will mean peaks that have been called in at least 2 replicates will contribute to the consensus set of peaks, and as such peaks that are unique to a given replicate will be discarded. + +```bash +-- min_reps_consensus 1 +``` + +### `--saveMACSPileup` +Instruct MACS2 to create bedGraph files using the `-B --SPMR` parameters. + +### `--skipDiffAnalysis` +Skip read counting and differential analysis step. + +## Skipping QC steps + +The pipeline contains a large number of quality control steps. Sometimes, it may not be desirable to run all of them if time and compute resources are limited. +The following options make this easy: + +| Step | Description | +|-------------------------|------------------------------------| +| `--skipFastQC` | Skip FastQC | +| `--skipPicardMetrics` | Skip Picard CollectMultipleMetrics | +| `--skipPreseq` | Skip Preseq | +| `--skipPlotProfile` | Skip deepTools plotProfile | +| `--skipPlotFingerprint` | Skip deepTools plotFingerprint | +| `--skipIGV` | Skip IGV | +| `--skipMultiQC` | Skip MultiQC | + +`--skipMultiQCStats` allows you to exclude the [general statistics table](https://multiqc.info/docs/#general-statistics-table) from the MultiQC report. + ## Job resources ### Automatic resubmission Each step in the pipeline has a default set of requirements for number of CPUs, memory and time. For most of the steps in the pipeline, if the job exits with an error code of `143` (exceeded requested resources) it will automatically resubmit with higher requests (2 x original, then 3 x original). If it still fails after three times then the pipeline is stopped. From 7f3df6f243956f5dc2d2dd5153e9ebd1ab20afb4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 4 Oct 2019 17:56:33 +0100 Subject: [PATCH 049/149] Add in deepTools and preseq --- bin/scrape_software_versions.py | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index 456c8fae..03e5b169 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -12,7 +12,7 @@ 'Samtools': ['v_samtools.txt', r"samtools (\S+)"], 'BEDTools': ['v_bedtools.txt', r"bedtools v(\S+)"], 'BamTools': ['v_bamtools.txt', r"bamtools (\S+)"], - #'deepTools': ['v_deeptools.txt', r"plotFingerprint (\S+)"], + 'deepTools': ['v_deeptools.txt', r"plotFingerprint (\S+)"], 'Picard': ['v_picard.txt', r"([\d\.]+)-SNAPSHOT"], 'R': ['v_R.txt', r"R version (\S+)"], 'Pysam': ['v_pysam.txt', r"(\S+)"], @@ -20,7 +20,7 @@ 'HOMER': ['v_homer.txt', r"(\S+)"], 'ataqv': ['v_ataqv.txt', r"(\S+)"], 'featureCounts': ['v_featurecounts.txt', r"featureCounts v(\S+)"], - #'Preseq': ['v_preseq.txt', r"Version: (\S+)"], + 'Preseq': ['v_preseq.txt', r"Version: (\S+)"], 'MultiQC': ['v_multiqc.txt', r"multiqc, version (\S+)"], } @@ -33,7 +33,7 @@ results['Samtools'] = 'N/A' results['BEDTools'] = 'N/A' results['BamTools'] = 'N/A' -#results['deepTools'] = 'N/A' +results['deepTools'] = 'N/A' results['Picard'] = 'N/A' results['R'] = 'N/A' results['Pysam'] = 'N/A' @@ -41,7 +41,7 @@ results['HOMER'] = False results['ataqv'] = 'N/A' results['featureCounts'] = 'N/A' -#results['Preseq'] = 'N/A' +results['Preseq'] = 'N/A' results['MultiQC'] = 'N/A' # Search each file using its regex From e3681d6147afe94f572f2ad3616393399d92d944 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 4 Oct 2019 17:56:42 +0100 Subject: [PATCH 050/149] Add in deepTools and preseq --- main.nf | 246 ++++++++++++++++++++++++++++---------------------------- 1 file changed, 123 insertions(+), 123 deletions(-) diff --git a/main.nf b/main.nf index f7bde6c8..bcf8edb9 100644 --- a/main.nf +++ b/main.nf @@ -840,32 +840,32 @@ if (params.singleEnd) { /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// -// /* -// * STEP 5.1 preseq analysis after merging libraries and before filtering -// */ -// process MergeLibraryBAMPreseq { -// tag "$name" -// label 'process_low' -// publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' -// -// when: -// !params.skipPreseq -// -// input: -// set val(name), file(bam) from ch_mlib_bam_preseq -// -// output: -// file "*.ccurve.txt" into ch_mlib_preseq_mqc -// -// script: -// prefix = "${name}.mLb.mkD" -// """ -// preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} -// """ -// } +/* + * STEP 5.1 preseq analysis after merging libraries and before filtering + */ +process MergeLibraryBAMPreseq { + tag "$name" + label 'process_low' + publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' + + when: + !params.skipPreseq + + input: + set val(name), file(bam) from ch_mlib_bam_preseq + + output: + file "*.ccurve.txt" into ch_mlib_preseq_mqc + + script: + prefix = "${name}.mLb.mkD" + """ + preseq lc_extrap -v -output ${prefix}.ccurve.txt -bam ${bam[0]} + """ +} /* - * STEP 4.4.1 Picard CollectMultipleMetrics after merging libraries + * STEP 5.2 Picard CollectMultipleMetrics after merging libraries */ process MergeLibraryBAMMetrics { tag "$name" @@ -907,7 +907,7 @@ process MergeLibraryBAMMetrics { } /* - * STEP 4.4.2 Read depth normalised bigWig + * STEP 5.3 Read depth normalised bigWig */ process MergeLibraryBigWig { tag "$name" @@ -943,80 +943,80 @@ process MergeLibraryBigWig { """ } -// /* -// * STEP 5.4 generate gene body coverage plot with deepTools -// */ -// process MergeLibraryPlotProfile { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' -// -// when: -// !params.skipPlotProfile -// -// input: -// set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile -// file bed from ch_gene_bed -// -// output: -// file '*.{gz,pdf}' into ch_mlib_plotprofile_results -// file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// """ -// computeMatrix scale-regions \\ -// --regionsFileName $bed \\ -// --scoreFileName $bigwig \\ -// --outFileName ${prefix}.computeMatrix.mat.gz \\ -// --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ -// --regionBodyLength 1000 \\ -// --beforeRegionStartLength 3000 \\ -// --afterRegionStartLength 3000 \\ -// --skipZeros \\ -// --smartLabels \\ -// -p $task.cpus -// -// plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ -// --outFileName ${prefix}.plotProfile.pdf \\ -// --outFileNameData ${prefix}.plotProfile.tab -// """ -// } - -// /* -// * STEP 6.1 deepTools plotFingerprint -// */ -// process MergeLibraryPlotFingerprint { -// tag "$name" -// label 'process_high' -// publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' -// -// when: -// !params.skipPlotFingerprint -// -// input: -// set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint -// -// output: -// file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results -// file '*.raw.txt' into ch_mlib_plotfingerprint_mqc -// -// script: -// prefix = "${name}.mLb.clN" -// extend = (params.singleEnd && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' -// """ -// plotFingerprint \\ -// --bamfiles ${bam[0]} \\ -// --plotFile ${prefix}.plotFingerprint.pdf \\ -// $extend \\ -// --labels $prefix \\ -// --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ -// --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ -// --skipZeros \\ -// --numberOfProcessors ${task.cpus} \\ -// --numberOfSamples ${params.fingerprint_bins} -// """ -// } +/* + * STEP 5.4 generate gene body coverage plot with deepTools + */ +process MergeLibraryPlotProfile { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotProfile", mode: 'copy' + + when: + !params.skipPlotProfile + + input: + set val(name), file(bigwig) from ch_mlib_bigwig_plotprofile + file bed from ch_gene_bed + + output: + file '*.{gz,pdf}' into ch_mlib_plotprofile_results + file '*.plotProfile.tab' into ch_mlib_plotprofile_mqc + + script: + prefix = "${name}.mLb.clN" + """ + computeMatrix scale-regions \\ + --regionsFileName $bed \\ + --scoreFileName $bigwig \\ + --outFileName ${prefix}.computeMatrix.mat.gz \\ + --outFileNameMatrix ${prefix}.computeMatrix.vals.mat.gz \\ + --regionBodyLength 1000 \\ + --beforeRegionStartLength 3000 \\ + --afterRegionStartLength 3000 \\ + --skipZeros \\ + --smartLabels \\ + -p $task.cpus + + plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ + --outFileName ${prefix}.plotProfile.pdf \\ + --outFileNameData ${prefix}.plotProfile.tab + """ +} + +/* + * STEP 5.5 deepTools plotFingerprint + */ +process MergeLibraryPlotFingerprint { + tag "$name" + label 'process_high' + publishDir "${params.outdir}/bwa/mergedLibrary/deepTools/plotFingerprint", mode: 'copy' + + when: + !params.skipPlotFingerprint + + input: + set val(name), file(bam) from ch_mlib_rm_orphan_bam_plotfingerprint + + output: + file '*.{txt,pdf}' into ch_mlib_plotfingerprint_results + file '*.raw.txt' into ch_mlib_plotfingerprint_mqc + + script: + prefix = "${name}.mLb.clN" + extend = (params.singleEnd && params.fragment_size > 0) ? "--extendReads ${params.fragment_size}" : '' + """ + plotFingerprint \\ + --bamfiles ${bam[0]} \\ + --plotFile ${prefix}.plotFingerprint.pdf \\ + $extend \\ + --labels $prefix \\ + --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ + --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ + --skipZeros \\ + --numberOfProcessors ${task.cpus} \\ + --numberOfSamples ${params.fingerprint_bins} + """ +} /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// @@ -1027,7 +1027,7 @@ process MergeLibraryBigWig { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score + * STEP 6.1 Call peaks with MACS2 and calculate FRiP score */ process MergeLibraryMACSCallPeak { tag "$name" @@ -1083,7 +1083,7 @@ process MergeLibraryMACSCallPeak { } /* - * STEP 4.5.2 Annotate peaks with HOMER + * STEP 6.2 Annotate peaks with HOMER */ process MergeLibraryAnnotatePeaks { tag "$name" @@ -1115,7 +1115,7 @@ process MergeLibraryAnnotatePeaks { } /* - * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + * STEP 6.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergeLibraryPeakQC { label "process_medium" @@ -1154,7 +1154,7 @@ process MergeLibraryPeakQC { } /* - * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ process MergeLibraryCreateConsensusPeakSet { label 'process_long' @@ -1208,7 +1208,7 @@ process MergeLibraryCreateConsensusPeakSet { } /* - * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file + * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MergeLibraryAnnotateConsensusPeakSet { label "process_medium" @@ -1243,7 +1243,7 @@ process MergeLibraryAnnotateConsensusPeakSet { } /* - * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ process MergeLibraryDeseqConsensusPeakSet { label 'process_medium' @@ -1299,7 +1299,7 @@ process MergeLibraryDeseqConsensusPeakSet { } /* - * STEP 4.6.1 Run ataqv on BAM file and corresponding peaks + * STEP 6.7 Run ataqv on BAM file and corresponding peaks */ process MergeLibraryAtaqv { tag "$name" @@ -1333,7 +1333,7 @@ process MergeLibraryAtaqv { } /* - * STEP 4.6.2 run ataqv mkarv on all JSON files to render web app + * STEP 6.8 run ataqv mkarv on all JSON files to render web app */ process MergeLibraryAtaqvMkarv { label 'process_medium' @@ -1364,7 +1364,7 @@ process MergeLibraryAtaqvMkarv { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 5.1 Merge library BAM files across all replicates + * STEP 7 Merge library BAM files across all replicates */ ch_mlib_rm_orphan_bam_mrep.map { it -> [ it[0].split('_')[0..-2].join('_'), it[1] ] } .groupTuple(by: [0]) @@ -1452,7 +1452,7 @@ process MergeReplicateBAM { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 4.4.2 Read depth normalised bigWig + * STEP 8.1 Read depth normalised bigWig */ process MergeReplicateBigWig { tag "$name" @@ -1492,7 +1492,7 @@ process MergeReplicateBigWig { } /* - * STEP 4.5.1 Call peaks with MACS2 and calculate FRiP score + * STEP 8.2 Call peaks with MACS2 and calculate FRiP score */ process MergeReplicateMACSCallPeak { tag "$name" @@ -1547,7 +1547,7 @@ process MergeReplicateMACSCallPeak { } /* - * STEP 4.5.2 Annotate peaks with HOMER + * STEP 8.3 Annotate peaks with HOMER */ process MergeReplicateAnnotatePeaks { tag "$name" @@ -1579,7 +1579,7 @@ process MergeReplicateAnnotatePeaks { } /* - * STEP 4.5.3 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation + * STEP 8.4 Aggregated QC plots for peaks, FRiP and peak-to-gene annotation */ process MergeReplicatePeakQC { label "process_medium" @@ -1618,7 +1618,7 @@ process MergeReplicatePeakQC { } /* - * STEP 4.5.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection + * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ process MergeReplicateCreateConsensusPeakSet { label 'process_long' @@ -1671,7 +1671,7 @@ process MergeReplicateCreateConsensusPeakSet { } /* - * STEP 4.5.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file + * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ process MergeReplicateAnnotateConsensusPeakSet { label "process_medium" @@ -1706,7 +1706,7 @@ process MergeReplicateAnnotateConsensusPeakSet { } /* - * STEP 4.5.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 + * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ process MergeReplicateDeseqConsensusPeakSet { label 'process_medium' @@ -1770,7 +1770,7 @@ process MergeReplicateDeseqConsensusPeakSet { /////////////////////////////////////////////////////////////////////////////// /* - * STEP 6 - Create IGV session file + * STEP 9 - Create IGV session file */ process IGV { publishDir "${params.outdir}/igv/${peaktype}", mode: 'copy' @@ -1834,7 +1834,7 @@ process get_software_versions { samtools --version > v_samtools.txt bedtools --version > v_bedtools.txt echo \$(bamtools --version 2>&1) > v_bamtools.txt - #echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true + echo \$(plotFingerprint --version 2>&1) > v_deeptools.txt || true picard MarkDuplicates --version &> v_picard.txt || true echo \$(R --version 2>&1) > v_R.txt python -c "import pysam; print(pysam.__version__)" > v_pysam.txt @@ -1842,7 +1842,7 @@ process get_software_versions { touch v_homer.txt echo \$(ataqv --version 2>&1) > v_ataqv.txt echo \$(featureCounts -v 2>&1) > v_featurecounts.txt - #preseq &> v_preseq.txt + preseq &> v_preseq.txt multiqc --version > v_multiqc.txt scrape_software_versions.py &> software_versions_mqc.yaml """ @@ -1867,7 +1867,7 @@ ${summary.collect { k,v -> "
$k
${v ?: ' Date: Fri, 4 Oct 2019 18:06:29 +0100 Subject: [PATCH 051/149] Add skipAtaqv parameter --- docs/usage.md | 1 + nextflow.config | 1 + 2 files changed, 2 insertions(+) diff --git a/docs/usage.md b/docs/usage.md index 212f5ef5..2936af4c 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -372,6 +372,7 @@ The following options make this easy: | `--skipPreseq` | Skip Preseq | | `--skipPlotProfile` | Skip deepTools plotProfile | | `--skipPlotFingerprint` | Skip deepTools plotFingerprint | +| `--skipAtaqv` | Skip Ataqv | | `--skipIGV` | Skip IGV | | `--skipMultiQC` | Skip MultiQC | diff --git a/nextflow.config b/nextflow.config index df68009a..bc1d1f62 100755 --- a/nextflow.config +++ b/nextflow.config @@ -47,6 +47,7 @@ params { skipPreseq = false skipPlotProfile = false skipPlotFingerprint = false + skipAtaqv = false skipIGV = false skipMultiQC = false skipMultiQCStats = false From d12027eb221e4e5b61bbfaaafd814dbbb1496914 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 4 Oct 2019 18:06:37 +0100 Subject: [PATCH 052/149] Add skipAtaqv parameter --- main.nf | 24 ++++++++++++++++-------- 1 file changed, 16 insertions(+), 8 deletions(-) diff --git a/main.nf b/main.nf index bcf8edb9..ef2392f4 100644 --- a/main.nf +++ b/main.nf @@ -76,6 +76,7 @@ def helpMessage() { --skipPreseq Skip Preseq --skipPlotProfile Skip deepTools plotProfile --skipPlotFingerprint Skip deepTools plotFingerprint + --skipAtaqv Skip Ataqv --skipIGV Skip IGV --skipMultiQC Skip MultiQC --skipMultiQCStats Exclude general statistics table from MultiQC report @@ -255,6 +256,7 @@ if (params.skipPicardMetrics) summary['Skip Picard Metrics'] = 'Yes' if (params.skipPreseq) summary['Skip Preseq'] = 'Yes' if (params.skipPlotProfile) summary['Skip plotProfile'] = 'Yes' if (params.skipPlotFingerprint) summary['Skip plotFingerprint'] = 'Yes' +if (params.skipAtaqv) summary['Skip Ataqv'] = 'Yes' if (params.skipIGV) summary['Skip IGV'] = 'Yes' if (params.skipMultiQC) summary['Skip MultiQC'] = 'Yes' if (params.skipMultiQCStats) summary['Skip MultiQC Stats'] = 'Yes' @@ -843,7 +845,7 @@ if (params.singleEnd) { /* * STEP 5.1 preseq analysis after merging libraries and before filtering */ -process MergeLibraryBAMPreseq { +process MergeLibraryPreseq { tag "$name" label 'process_low' publishDir "${params.outdir}/bwa/mergedLibrary/preseq", mode: 'copy' @@ -867,7 +869,7 @@ process MergeLibraryBAMPreseq { /* * STEP 5.2 Picard CollectMultipleMetrics after merging libraries */ -process MergeLibraryBAMMetrics { +process MergeLibraryMetrics { tag "$name" label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', @@ -1156,7 +1158,7 @@ process MergeLibraryPeakQC { /* * STEP 6.4 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process MergeLibraryCreateConsensusPeakSet { +process MergeLibraryConsensusPeakSet { label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus", mode: 'copy', saveAs: {filename -> @@ -1210,7 +1212,7 @@ process MergeLibraryCreateConsensusPeakSet { /* * STEP 6.5 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process MergeLibraryAnnotateConsensusPeakSet { +process MergeLibraryConsensusPeakSetAnnotate { label "process_medium" publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus", mode: 'copy' @@ -1245,7 +1247,7 @@ process MergeLibraryAnnotateConsensusPeakSet { /* * STEP 6.6 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process MergeLibraryDeseqConsensusPeakSet { +process MergeLibraryConsensusPeakSetDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus/deseq2", mode: 'copy', saveAs: {filename -> @@ -1306,6 +1308,9 @@ process MergeLibraryAtaqv { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' + when: + !skipAtaqv + input: set val(name), file(bam), file(peak) from ch_mlib_bam_ataqv.join(ch_mlib_macs_ataqv, by: [0]) file autosomes from ch_genome_autosomes.collect() @@ -1339,6 +1344,9 @@ process MergeLibraryAtaqvMkarv { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/ataqv", mode: 'copy' + when: + !skipAtaqv + input: file json from ch_mlib_ataqv.collect() @@ -1620,7 +1628,7 @@ process MergeReplicatePeakQC { /* * STEP 8.5 Consensus peaks across samples, create boolean filtering file, .saf file for featureCounts and UpSetR plot for intersection */ -process MergeReplicateCreateConsensusPeakSet { +process MergeReplicateConsensusPeakSet { label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus", mode: 'copy', saveAs: {filename -> @@ -1673,7 +1681,7 @@ process MergeReplicateCreateConsensusPeakSet { /* * STEP 8.6 Annotate consensus peaks with HOMER, and add annotation to boolean output file */ -process MergeReplicateAnnotateConsensusPeakSet { +process MergeReplicateConsensusPeakSetAnnotate { label "process_medium" publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus", mode: 'copy' @@ -1708,7 +1716,7 @@ process MergeReplicateAnnotateConsensusPeakSet { /* * STEP 8.7 Count reads in consensus peaks with featureCounts and perform differential analysis with DESeq2 */ -process MergeReplicateDeseqConsensusPeakSet { +process MergeReplicateConsensusPeakSetDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus/deseq2", mode: 'copy', saveAs: {filename -> From 1f531b0fa0168c3753023e711fb5b5b8b862eba0 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Mon, 7 Oct 2019 10:40:47 +0100 Subject: [PATCH 053/149] Fix gene bed issue --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index ef2392f4..6f36ac7a 100644 --- a/main.nf +++ b/main.nf @@ -171,7 +171,7 @@ ch_mrep_deseq2_clustering_header = file("$baseDir/assets/multiqc/mrep_deseq2_clu // Validate inputs if (params.design) { ch_design = file(params.design, checkIfExists: true) } else { exit 1, "Samples design file not specified!" } if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } -if (params.gene_bed) { ch_gene_bed = file(params.gene.bed, checkIfExists: true) } +if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } From 18bb26da4ef48117b05141139ff1acf77055cdff Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:33:59 +0100 Subject: [PATCH 054/149] v1.7 update CONTRIBUTING.md --- .github/CONTRIBUTING.md | 6 +++--- 1 file changed, 3 insertions(+), 3 deletions(-) diff --git a/.github/CONTRIBUTING.md b/.github/CONTRIBUTING.md index 6376a3c9..da76fa43 100644 --- a/.github/CONTRIBUTING.md +++ b/.github/CONTRIBUTING.md @@ -6,7 +6,7 @@ We try to manage the required tasks for nf-core/atacseq using GitHub issues, you However, don't be put off by this template - other more general issues and suggestions are welcome! Contributions to the code are even more welcome ;) -> If you need help using or modifying nf-core/atacseq then the best place to ask is on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). +> If you need help using or modifying nf-core/atacseq then the best place to ask is on the pipeline channel on [Slack](https://nf-co.re/join/slack/). @@ -32,7 +32,7 @@ Typically, pull-requests are only fully reviewed when these tests are passing, t There are typically two types of tests that run: ### Lint Tests -The nf-core has a [set of guidelines](http://nf-co.re/guidelines) which all pipelines must adhere to. +The nf-core has a [set of guidelines](https://nf-co.re/developers/guidelines) which all pipelines must adhere to. To enforce these and ensure that all pipelines stay in sync, we have developed a helper tool which runs checks on the pipeline code. This is in the [nf-core/tools repository](https://github.com/nf-core/tools) and once installed can be run locally with the `nf-core lint ` command. If any failures or warnings are encountered, please follow the listed URL for more documentation. @@ -44,4 +44,4 @@ If there are any failures then the automated tests fail. These tests are run both with the latest available version of Nextflow and also the minimum required version that is stated in the pipeline code. ## Getting help -For further information/help, please consult the [nf-core/atacseq documentation](https://github.com/nf-core/atacseq#documentation) and don't hesitate to get in touch on the pipeline channel on [Slack](https://nf-core-invite.herokuapp.com/). +For further information/help, please consult the [nf-core/atacseq documentation](https://github.com/nf-core/atacseq#documentation) and don't hesitate to get in touch on the [nf-core/atacseq pipeline channel](https://nfcore.slack.com/channels/atacseq) on [Slack](https://nf-co.re/join/slack/). From 956310b83a5aa2b33bfd43f196b0f7493c5a1ecd Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:34:13 +0100 Subject: [PATCH 055/149] v1.7 update .travis.yml --- .travis.yml | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/.travis.yml b/.travis.yml index b00e5355..9409abbe 100755 --- a/.travis.yml +++ b/.travis.yml @@ -9,7 +9,7 @@ matrix: before_install: # PRs to master are only ok if coming from dev branch - - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && [ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ])' + - '[ $TRAVIS_PULL_REQUEST = "false" ] || [ $TRAVIS_BRANCH != "master" ] || ([ $TRAVIS_PULL_REQUEST_SLUG = $TRAVIS_REPO_SLUG ] && ([ $TRAVIS_PULL_REQUEST_BRANCH = "dev" ] || [ $TRAVIS_PULL_REQUEST_BRANCH = "patch" ]))' # Pull the docker image first so the test doesn't wait for this - docker pull nfcore/atacseq:dev # Fake the tag locally so that the pipeline runs properly From 246bb819fe0fdf868deaf34af2ff362c86c03e9c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:34:27 +0100 Subject: [PATCH 056/149] v1.7 update CODE_OF_CONDUCT.md --- CODE_OF_CONDUCT.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/CODE_OF_CONDUCT.md b/CODE_OF_CONDUCT.md index 09226d0d..1cda7600 100755 --- a/CODE_OF_CONDUCT.md +++ b/CODE_OF_CONDUCT.md @@ -34,7 +34,7 @@ This Code of Conduct applies both within project spaces and in public spaces whe ## Enforcement -Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-core-invite.herokuapp.com/). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. +Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team on [Slack](https://nf-co.re/join/slack/). The project team will review and investigate all complaints, and will respond in a way that it deems appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. Project maintainers who do not follow or enforce the Code of Conduct in good faith may face temporary or permanent repercussions as determined by other members of the project's leadership. From 467452705f1ef5aa439b1d75d66f794221571be9 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:34:40 +0100 Subject: [PATCH 057/149] v1.7 update Dockerfile --- Dockerfile | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/Dockerfile b/Dockerfile index c8a96fc1..31b34ca3 100755 --- a/Dockerfile +++ b/Dockerfile @@ -1,4 +1,4 @@ -FROM nfcore/base +FROM nfcore/base:1.7 LABEL authors="Harshil Patel" \ description="Docker image containing all requirements for nf-core/atacseq pipeline" From 8f6d0a25acb243d21525e9cf52e64b3a2d5d2af4 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:34:53 +0100 Subject: [PATCH 058/149] v1.7 update README --- README.md | 12 +++++++++--- 1 file changed, 9 insertions(+), 3 deletions(-) diff --git a/README.md b/README.md index 391ffa41..443cb2b5 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,4 @@ -# ![nfcore/atacseq](docs/images/nfcore-atacseq_logo.png) +# ![nfcore/atacseq](docs/images/nf-core-atacseq_logo.png) [![Build Status](https://travis-ci.org/nf-core/atacseq.svg?branch=master)](https://travis-ci.org/nf-core/atacseq) [![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A50.32.0-brightgreen.svg)](https://www.nextflow.io/) @@ -96,11 +96,17 @@ The pipeline was developed by [Harshil Patel](mailto:harshil.patel@crick.ac.uk). The [nf-core/rnaseq](https://github.com/nf-core/rnaseq) and [nf-core/chipseq](https://github.com/nf-core/chipseq) pipelines developed by Phil Ewels were initially used as a template for this pipeline. Many thanks to Phil for all of his help and advice, and the team at SciLifeLab. -Many thanks to others who have helped out along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@sven1103](https://github.com/sven1103), [@MaxUlysse](https://github.com/MaxUlysse), [@micans](https://github.com/micans), [@pditommaso](https://github.com/pditommaso). +Many thanks to others who have helped out along the way too, including (but not limited to): [@apeltzer](https://github.com/apeltzer), [@sven1103](https://github.com/sven1103), [@MaxUlysse](https://github.com/MaxUlysse), [@micans](https://github.com/micans), [@jinmingda](https://github.com/jinmingda), [@ktrns](https://github.com/ktrns), [@crickbabs](https://github.com/crickbabs), [@pditommaso](https://github.com/pditommaso). + +## Contributions and Support + +If you would like to contribute to this pipeline, please see the [contributing guidelines](.github/CONTRIBUTING.md). + +For further information or help, don't hesitate to get in touch on [Slack](https://nfcore.slack.com/channels/atacseq) (you can join with [this invite](https://nf-co.re/join/slack)). ## Citation If you use nf-core/atacseq for your analysis, please cite it using the following doi: [10.5281/zenodo.2634132](https://doi.org/10.5281/zenodo.2634132) You can cite the `nf-core` pre-print as follows: -Ewels PA, Peltzer A, Fillinger S, Alneberg JA, Patel H, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. **nf-core: Community curated bioinformatics pipelines**. *bioRxiv*. 2019. p. 610741. [doi: 10.1101/610741](https://www.biorxiv.org/content/10.1101/610741v1). +> Ewels PA, Peltzer A, Fillinger S, Alneberg JA, Patel H, Wilm A, Garcia MU, Di Tommaso P, Nahnsen S. **nf-core: Community curated bioinformatics pipelines**. *bioRxiv*. 2019. p. 610741. [doi: 10.1101/610741](https://www.biorxiv.org/content/10.1101/610741v1). From 849fb6bd75c66955fee0819a546e6e3f6e5d1984 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:35:29 +0100 Subject: [PATCH 059/149] v1.7 update email files --- assets/email_template.html | 2 +- assets/email_template.txt | 12 +++++++++--- assets/sendmail_template.txt | 8 ++++---- 3 files changed, 14 insertions(+), 8 deletions(-) diff --git a/assets/email_template.html b/assets/email_template.html index ecab0e13..61a98cb6 100755 --- a/assets/email_template.html +++ b/assets/email_template.html @@ -11,7 +11,7 @@
- +

nf-core/atacseq v${version}

Run Name: $runName

diff --git a/assets/email_template.txt b/assets/email_template.txt index 91d3a7a6..5f89db73 100755 --- a/assets/email_template.txt +++ b/assets/email_template.txt @@ -1,6 +1,12 @@ -======================================== - nf-core/atacseq v${version} -======================================== +---------------------------------------------------- + ,--./,-. + ___ __ __ __ ___ /,-._.--~\\ + |\\ | |__ __ / ` / \\ |__) |__ } { + | \\| | \\__, \\__/ | \\ |___ \\`-._,-`-, + `._,._,' + nf-core/atacseq v${version} +---------------------------------------------------- + Run Name: $runName <% if (success){ diff --git a/assets/sendmail_template.txt b/assets/sendmail_template.txt index 3777e08a..fef351fd 100755 --- a/assets/sendmail_template.txt +++ b/assets/sendmail_template.txt @@ -9,12 +9,12 @@ Content-Type: text/html; charset=utf-8 $email_html --nfcoremimeboundary -Content-Type: image/png;name="nfcore-atacseq_logo.png" +Content-Type: image/png;name="nf-core-atacseq_logo.png" Content-Transfer-Encoding: base64 -Content-ID: -Content-Disposition: inline; filename="nfcore-atacseq_logo.png" +Content-ID: +Content-Disposition: inline; filename="nf-core-atacseq_logo.png" -<% out << new File("$baseDir/assets/nfcore-atacseq_logo.png"). +<% out << new File("$baseDir/assets/nf-core-atacseq_logo.png"). bytes. encodeBase64(). toString(). From 4b6e2fa1c1475fcaef2e095b84dc02855615cb9e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:36:00 +0100 Subject: [PATCH 060/149] v1.7 update configs --- conf/base.config | 17 +++++++++-------- 1 file changed, 9 insertions(+), 8 deletions(-) diff --git a/conf/base.config b/conf/base.config index 48ac17fa..7e49cc37 100755 --- a/conf/base.config +++ b/conf/base.config @@ -12,8 +12,8 @@ process { cpus = { check_max( 1 * task.attempt, 'cpus' ) } - memory = { check_max( 8.GB * task.attempt, 'memory' ) } - time = { check_max( 2.h * task.attempt, 'time' ) } + memory = { check_max( 7.GB * task.attempt, 'memory' ) } + time = { check_max( 4.h * task.attempt, 'time' ) } errorStrategy = { task.exitStatus in [143,137,104,134,139] ? 'retry' : 'finish' } maxRetries = 1 @@ -22,29 +22,30 @@ process { // Process-specific resource requirements withLabel:process_low { cpus = { check_max( 2 * task.attempt, 'cpus' ) } - memory = { check_max( 16.GB * task.attempt, 'memory' ) } - time = { check_max( 4.h * task.attempt, 'time' ) } + memory = { check_max( 14.GB * task.attempt, 'memory' ) } + time = { check_max( 6.h * task.attempt, 'time' ) } } withLabel:process_medium { cpus = { check_max( 6 * task.attempt, 'cpus' ) } - memory = { check_max( 48.GB * task.attempt, 'memory' ) } + memory = { check_max( 42.GB * task.attempt, 'memory' ) } time = { check_max( 8.h * task.attempt, 'time' ) } } withLabel:process_high { cpus = { check_max( 12 * task.attempt, 'cpus' ) } - memory = { check_max( 96.GB * task.attempt, 'memory' ) } + memory = { check_max( 84.GB * task.attempt, 'memory' ) } time = { check_max( 16.h * task.attempt, 'time' ) } } withLabel:process_long { - time = { check_max( 12.h * task.attempt, 'time' ) } + time = { check_max( 20.h * task.attempt, 'time' ) } } withName:get_software_versions { cache = false } + } -// Defaults config params, may be overwritten by later configs params { + // Defaults only, expecting to be overwritten max_memory = 128.GB max_cpus = 16 max_time = 240.h From 2a85a6e5e92bac9be2991d34ca890d689377d63e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:36:09 +0100 Subject: [PATCH 061/149] v1.7 update igenomes --- conf/igenomes.config | 219 ++++++++++++++++++++++++++++++++++++++++--- 1 file changed, 204 insertions(+), 15 deletions(-) diff --git a/conf/igenomes.config b/conf/igenomes.config index 3c36daa2..37217cf4 100755 --- a/conf/igenomes.config +++ b/conf/igenomes.config @@ -13,33 +13,45 @@ params { 'GRCh37' { fasta = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/GRCh37-blacklist.bed" - macs_gsize = "2.7e9" mito_name = "MT" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/GRCh37-blacklist.bed" } 'GRCh38' { fasta = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/NCBI/GRCh38/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" - macs_gsize = "2.7e9" mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" } 'GRCm38' { fasta = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Mus_musculus/Ensembl/GRCm38/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/GRCm38-blacklist.bed" - macs_gsize = "1.87e9" mito_name = "MT" + macs_gsize = "1.87e9" + blacklist = "${baseDir}/assets/blacklists/GRCm38-blacklist.bed" } 'TAIR10' { fasta = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Arabidopsis_thaliana/Ensembl/TAIR10/Annotation/Genes/genes.bed" mito_name = "Mt" @@ -47,12 +59,18 @@ params { 'EB2' { fasta = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Bacillus_subtilis_168/Ensembl/EB2/Annotation/Genes/genes.bed" } 'UMD3.1' { fasta = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Bos_taurus/Ensembl/UMD3.1/Annotation/Genes/genes.bed" mito_name = "MT" @@ -60,14 +78,20 @@ params { 'WBcel235' { fasta = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/Ensembl/WBcel235/Annotation/Genes/genes.bed" - macs_gsize = "9e7" mito_name = "MtDNA" + macs_gsize = "9e7" } 'CanFam3.1' { fasta = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Canis_familiaris/Ensembl/CanFam3.1/Annotation/Genes/genes.bed" mito_name = "MT" @@ -75,6 +99,9 @@ params { 'GRCz10' { fasta = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Danio_rerio/Ensembl/GRCz10/Annotation/Genes/genes.bed" mito_name = "MT" @@ -82,14 +109,20 @@ params { 'BDGP6' { fasta = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Drosophila_melanogaster/Ensembl/BDGP6/Annotation/Genes/genes.bed" - macs_gsize = "1.2e8" mito_name = "M" + macs_gsize = "1.2e8" } 'EquCab2' { fasta = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Equus_caballus/Ensembl/EquCab2/Annotation/Genes/genes.bed" mito_name = "MT" @@ -97,12 +130,18 @@ params { 'EB1' { fasta = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Escherichia_coli_K_12_DH10B/Ensembl/EB1/Annotation/Genes/genes.bed" } 'Galgal4' { fasta = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Gallus_gallus/Ensembl/Galgal4/Annotation/Genes/genes.bed" mito_name = "MT" @@ -110,12 +149,18 @@ params { 'Gm01' { fasta = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Glycine_max/Ensembl/Gm01/Annotation/Genes/genes.bed" } 'Mmul_1' { fasta = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Macaca_mulatta/Ensembl/Mmul_1/Annotation/Genes/genes.bed" mito_name = "MT" @@ -123,6 +168,9 @@ params { 'IRGSP-1.0' { fasta = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Oryza_sativa_japonica/Ensembl/IRGSP-1.0/Annotation/Genes/genes.bed" mito_name = "Mt" @@ -130,6 +178,9 @@ params { 'CHIMP2.1.4' { fasta = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Pan_troglodytes/Ensembl/CHIMP2.1.4/Annotation/Genes/genes.bed" mito_name = "MT" @@ -137,6 +188,9 @@ params { 'Rnor_6.0' { fasta = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Rattus_norvegicus/Ensembl/Rnor_6.0/Annotation/Genes/genes.bed" mito_name = "MT" @@ -144,28 +198,40 @@ params { 'R64-1-1' { fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Saccharomyces_cerevisiae/Ensembl/R64-1-1/Annotation/Genes/genes.bed" - macs_gsize = "1.2e7" mito_name = "MT" + macs_gsize = "1.2e7" } 'EF2' { fasta = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Schizosaccharomyces_pombe/Ensembl/EF2/Annotation/Genes/genes.bed" - macs_gsize = "1.21e7" mito_name = "MT" + macs_gsize = "1.21e7" } 'Sbi1' { fasta = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Sorghum_bicolor/Ensembl/Sbi1/Annotation/Genes/genes.bed" } 'Sscrofa10.2' { fasta = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Sus_scrofa/Ensembl/Sscrofa10.2/Annotation/Genes/genes.bed" mito_name = "MT" @@ -173,6 +239,9 @@ params { 'AGPv3' { fasta = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Zea_mays/Ensembl/AGPv3/Annotation/Genes/genes.bed" mito_name = "Mt" @@ -180,28 +249,148 @@ params { 'hg38' { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg38/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" - macs_gsize = "2.7e9" mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg38-blacklist.bed" } 'hg19' { fasta = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Homo_sapiens/UCSC/hg19/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/hg19-blacklist.bed" - macs_gsize = "2.7e9" mito_name = "chrM" + macs_gsize = "2.7e9" + blacklist = "${baseDir}/assets/blacklists/hg19-blacklist.bed" } 'mm10' { fasta = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/WholeGenomeFasta/genome.fa" bwa = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Sequence/BismarkIndex/" gtf = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.gtf" bed12 = "${params.igenomes_base}/Mus_musculus/UCSC/mm10/Annotation/Genes/genes.bed" - blacklist = "${baseDir}/assets/blacklists/mm10-blacklist.bed" + mito_name = "chrM" macs_gsize = "1.87e9" + blacklist = "${baseDir}/assets/blacklists/mm10-blacklist.bed" + } + 'bosTau8' { + fasta = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Bos_taurus/UCSC/bosTau8/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'ce10' { + fasta = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Caenorhabditis_elegans/UCSC/ce10/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "9e7" + } + 'canFam3' { + fasta = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Canis_familiaris/UCSC/canFam3/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'danRer10' { + fasta = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Danio_rerio/UCSC/danRer10/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'dm6' { + fasta = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Drosophila_melanogaster/UCSC/dm6/Annotation/Genes/genes.bed" + mito_name = "chrM" + macs_gsize = "1.2e8" + } + 'equCab2' { + fasta = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Equus_caballus/UCSC/equCab2/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'galGal4' { + fasta = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Gallus_gallus/UCSC/galGal4/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'panTro4' { + fasta = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Pan_troglodytes/UCSC/panTro4/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'rn6' { + fasta = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Rattus_norvegicus/UCSC/rn6/Annotation/Genes/genes.bed" + mito_name = "chrM" + } + 'sacCer3' { + fasta = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Saccharomyces_cerevisiae/UCSC/sacCer3/Sequence/BismarkIndex/" + mito_name = "chrM" + macs_gsize = "1.2e7" + } + 'susScr3' { + fasta = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/WholeGenomeFasta/genome.fa" + bwa = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BWAIndex/genome.fa" + bowtie2 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/Bowtie2Index/" + star = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/STARIndex/" + bismark = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Sequence/BismarkIndex/" + gtf = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.gtf" + bed12 = "${params.igenomes_base}/Sus_scrofa/UCSC/susScr3/Annotation/Genes/genes.bed" mito_name = "chrM" } } From 5857ab6c887e5ec5f2ab40f9fd4737cd1683aa99 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:36:28 +0100 Subject: [PATCH 062/149] v1.7 update software versions --- bin/scrape_software_versions.py | 15 +++++++++------ 1 file changed, 9 insertions(+), 6 deletions(-) diff --git a/bin/scrape_software_versions.py b/bin/scrape_software_versions.py index 03e5b169..0b76c7a6 100755 --- a/bin/scrape_software_versions.py +++ b/bin/scrape_software_versions.py @@ -46,12 +46,15 @@ # Search each file using its regex for k, v in regexes.items(): - with open(v[0]) as x: - versions = x.read() - match = re.search(v[1], versions) - if match: - results[k] = "v{}".format(match.group(1)) - + try: + with open(v[0]) as x: + versions = x.read() + match = re.search(v[1], versions) + if match: + results[k] = "v{}".format(match.group(1)) + except IOError: + results[k] = False + # Remove software set to false in results for k in results: if not results[k]: From 4de0746f05da967a1fdcb38d5d42f20c999247e7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:36:39 +0100 Subject: [PATCH 063/149] v1.7 update docs --- docs/usage.md | 8 ++++++-- 1 file changed, 6 insertions(+), 2 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index 2936af4c..fe069a78 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -54,6 +54,7 @@ * [Other command line parameters](#other-command-line-parameters) * [`--outdir`](#--outdir) * [`--email`](#--email) + * [`--email_on_fail`](#--email_on_fail) * [`-name`](#-name) * [`-resume`](#-resume) * [`-c`](#-c) @@ -113,7 +114,7 @@ This version number will be logged in reports when you run the pipeline, so that ## Main arguments ### `-profile` -Use this parameter to choose a configuration profile. Profiles can give configuration pre-sets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! +Use this parameter to choose a configuration profile. Profiles can give configuration presets for different compute environments. Note that multiple profiles can be loaded, for example: `-profile docker` - the order of arguments is important! If `-profile` is not specified at all the pipeline will be run locally and expects all software to be installed and available on the `PATH`. @@ -387,7 +388,7 @@ Wherever process-specific requirements are set in the pipeline, the default valu If you are likely to be running `nf-core` pipelines regularly it may be a good idea to request that your custom config file is uploaded to the `nf-core/configs` git repository. Before you do this please can you test that the config file works with your pipeline of choice using the `-c` parameter (see definition below). You can then create a pull request to the `nf-core/configs` repository with the addition of your config file, associated documentation file (see examples in [`nf-core/configs/docs`](https://github.com/nf-core/configs/tree/master/docs)), and amending [`nfcore_custom.config`](https://github.com/nf-core/configs/blob/master/nfcore_custom.config) to include your custom profile. -If you have any questions or issues please send us a message on [Slack](https://nf-core-invite.herokuapp.com/). +If you have any questions or issues please send us a message on [Slack](https://nf-co.re/join/slack/). ## AWS Batch specific parameters Running the pipeline on AWS Batch requires a couple of specific parameters to be set according to your AWS Batch configuration. Please use the `-awsbatch` profile and then specify all of the following parameters. @@ -406,6 +407,9 @@ The output directory where the results will be saved. ### `--email` Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits. If set in your user config file (`~/.nextflow/config`) then you don't need to specify this on the command line for every run. +### `--email_on_fail` +This works exactly as with `--email`, except emails are only sent if the workflow is not successful. + ### `-name` Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. From 4be2a575829b0c121eb4b353d31491b545a1df2a Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:36:51 +0100 Subject: [PATCH 064/149] v1.7 update main script --- main.nf | 39 ++++++++++++++++++++++++--------------- 1 file changed, 24 insertions(+), 15 deletions(-) diff --git a/main.nf b/main.nf index 6f36ac7a..80a73252 100644 --- a/main.nf +++ b/main.nf @@ -84,6 +84,7 @@ def helpMessage() { Other --outdir The output directory where the results will be saved --email Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits + --email_on_fail Same as --email, except only send mail if the workflow is not successful --maxMultiqcEmailFileSize Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) -name Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic @@ -275,9 +276,10 @@ summary['Config Profile'] = workflow.profile if (params.config_profile_description) summary['Config Description'] = params.config_profile_description if (params.config_profile_contact) summary['Config Contact'] = params.config_profile_contact if (params.config_profile_url) summary['Config URL'] = params.config_profile_url -if (params.email) { - summary['E-mail Address'] = params.email - summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize +if (params.email || params.email_on_fail) { + summary['E-mail Address'] = params.email + summary['E-mail on failure'] = params.email_on_fail + summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize } log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join("\n") log.info "\033[2m----------------------------------------------------\033[0m" @@ -1993,11 +1995,12 @@ workflow.onComplete { email_fields['summary']['Nextflow Build'] = workflow.nextflow.build email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp + // TODO nf-core: If not using MultiQC, strip out this code (including params.maxMultiqcEmailFileSize) // On success try attach the multiqc report def mqc_report = null try { if (workflow.success) { - mqc_report = ch_multiqc_report.getVal() + mqc_report = multiqc_report.getVal() if (mqc_report.getClass() == ArrayList) { log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" mqc_report = mqc_report[0] @@ -2007,6 +2010,12 @@ workflow.onComplete { log.warn "[nf-core/atacseq] Could not attach MultiQC report to summary email" } + // Check if we are only sending emails on failure + email_address = params.email + if (!params.email && params.email_on_fail && !workflow.success) { + email_address = params.email_on_fail + } + // Render the TXT template def engine = new groovy.text.GStringTemplateEngine() def tf = new File("$baseDir/assets/email_template.txt") @@ -2019,29 +2028,29 @@ workflow.onComplete { def email_html = html_template.toString() // Render the sendmail template - def smail_fields = [ email: params.email, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.maxMultiqcEmailFileSize.toBytes() ] + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.maxMultiqcEmailFileSize.toBytes() ] def sf = new File("$baseDir/assets/sendmail_template.txt") def sendmail_template = engine.createTemplate(sf).make(smail_fields) def sendmail_html = sendmail_template.toString() // Send the HTML e-mail - if (params.email) { + if (email_address) { try { - if ( params.plaintext_email ) { throw GroovyException('Send plaintext e-mail, not HTML') } + if ( params.plaintext_email ){ throw GroovyException('Send plaintext e-mail, not HTML') } // Try to send HTML e-mail using sendmail [ 'sendmail', '-t' ].execute() << sendmail_html - log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (sendmail)" + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" } catch (all) { // Catch failures and try with plaintext - [ 'mail', '-s', subject, params.email ].execute() << email_txt - log.info "[nf-core/atacseq] Sent summary e-mail to $params.email (mail)" + [ 'mail', '-s', subject, email_address ].execute() << email_txt + log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (mail)" } } // Write summary e-mail HTML to a file def output_d = new File( "${params.outdir}/pipeline_info/" ) - if ( !output_d.exists() ) { - output_d.mkdirs() + if (!output_d.exists()) { + output_d.mkdirs() } def output_hf = new File( output_d, "pipeline_report.html" ) output_hf.withWriter { w -> w << email_html } @@ -2076,7 +2085,7 @@ workflow.onComplete { /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// -def nfcoreHeader() { +def nfcoreHeader(){ // Log colors ANSI codes c_reset = params.monochrome_logs ? '' : "\033[0m"; c_dim = params.monochrome_logs ? '' : "\033[2m"; @@ -2088,14 +2097,14 @@ def nfcoreHeader() { c_cyan = params.monochrome_logs ? '' : "\033[0;36m"; c_white = params.monochrome_logs ? '' : "\033[0;37m"; - return """ ${c_dim}----------------------------------------------------${c_reset} + return """ -${c_dim}--------------------------------------------------${c_reset}- ${c_green},--.${c_black}/${c_green},-.${c_reset} ${c_blue} ___ __ __ __ ___ ${c_green}/,-._.--~\'${c_reset} ${c_blue} |\\ | |__ __ / ` / \\ |__) |__ ${c_yellow}} {${c_reset} ${c_blue} | \\| | \\__, \\__/ | \\ |___ ${c_green}\\`-._,-`-,${c_reset} ${c_green}`._,._,\'${c_reset} ${c_purple} nf-core/atacseq v${workflow.manifest.version}${c_reset} - ${c_dim}----------------------------------------------------${c_reset} + -${c_dim}--------------------------------------------------${c_reset}- """.stripIndent() } From 3284db16022822843218ffbdb08d98602173322d Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:37:03 +0100 Subject: [PATCH 065/149] v1.7 update nextflow.config --- nextflow.config | 44 ++++++++++++++++++++++++-------------------- 1 file changed, 24 insertions(+), 20 deletions(-) diff --git a/nextflow.config b/nextflow.config index bc1d1f62..0de69181 100755 --- a/nextflow.config +++ b/nextflow.config @@ -52,15 +52,6 @@ params { skipMultiQC = false skipMultiQCStats = false - // Options: Other - outdir = './results' - igenomes_base = "./iGenomes" - email = false - plaintext_email = false - monochrome_logs = false - name = false - maxMultiqcEmailFileSize = 25.MB - // Options: AWSBatch awsqueue = false awsregion = 'eu-west-1' @@ -71,18 +62,26 @@ params { bamtools_filter_se_config = "$baseDir/assets/bamtools_filter_se.json" // Options: Custom config + custom_config_version = 'master' + custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" config_profile_description = false config_profile_contact = false config_profile_url = false - // Options: Default + // Options: Other help = false - hostnames = false + outdir = './results' + igenomes_base = "./iGenomes" + igenomesIgnore = false + maxMultiqcEmailFileSize = 25.MB tracedir = "${params.outdir}/pipeline_info" + email = false + email_on_fail = false + plaintext_email = false + monochrome_logs = false + name = false + hostnames = false clusterOptions = false - igenomesIgnore = false - custom_config_version = 'master' - custom_config_base = "https://raw.githubusercontent.com/nf-core/configs/${params.custom_config_version}" } @@ -109,8 +108,13 @@ profiles { test { includeConfig 'conf/test.config' } } +// Avoid this error: +// WARNING: Your kernel does not support swap limit capabilities or the cgroup is not mounted. Memory limited without swap. +// Testing this in nf-core after discussion here https://github.com/nf-core/tools/pull/351, once this is established and works well, nextflow might implement this behavior as new default. +docker.runOptions = '-u \$(id -u):\$(id -g)' + // Load igenomes.config if required -if(!params.igenomesIgnore){ +if (!params.igenomesIgnore) { includeConfig 'conf/igenomes.config' } @@ -150,9 +154,9 @@ manifest { // Function to ensure that resource requirements don't go beyond // a maximum limit def check_max(obj, type) { - if(type == 'memory'){ + if (type == 'memory') { try { - if(obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) + if (obj.compareTo(params.max_memory as nextflow.util.MemoryUnit) == 1) return params.max_memory as nextflow.util.MemoryUnit else return obj @@ -160,9 +164,9 @@ def check_max(obj, type) { println " ### ERROR ### Max memory '${params.max_memory}' is not valid! Using default value: $obj" return obj } - } else if(type == 'time'){ + } else if (type == 'time') { try { - if(obj.compareTo(params.max_time as nextflow.util.Duration) == 1) + if (obj.compareTo(params.max_time as nextflow.util.Duration) == 1) return params.max_time as nextflow.util.Duration else return obj @@ -170,7 +174,7 @@ def check_max(obj, type) { println " ### ERROR ### Max time '${params.max_time}' is not valid! Using default value: $obj" return obj } - } else if(type == 'cpus'){ + } else if (type == 'cpus') { try { return Math.min( obj, params.max_cpus as int ) } catch (all) { From ddbd9048c9850b262518615d18f60a2178f4c08e Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 16:37:43 +0100 Subject: [PATCH 066/149] Update logo names --- ...re-atacseq_logo.png => nf-core-atacseq_logo.png} | Bin ...re-atacseq_logo.png => nf-core-atacseq_logo.png} | Bin 2 files changed, 0 insertions(+), 0 deletions(-) rename assets/{nfcore-atacseq_logo.png => nf-core-atacseq_logo.png} (100%) rename docs/images/{nfcore-atacseq_logo.png => nf-core-atacseq_logo.png} (100%) diff --git a/assets/nfcore-atacseq_logo.png b/assets/nf-core-atacseq_logo.png similarity index 100% rename from assets/nfcore-atacseq_logo.png rename to assets/nf-core-atacseq_logo.png diff --git a/docs/images/nfcore-atacseq_logo.png b/docs/images/nf-core-atacseq_logo.png similarity index 100% rename from docs/images/nfcore-atacseq_logo.png rename to docs/images/nf-core-atacseq_logo.png From 32d3112e2a013d5d33b173c54541518a921b5383 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Thu, 10 Oct 2019 17:02:18 +0100 Subject: [PATCH 067/149] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 7cf59f03..df6c88a3 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,6 +24,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * `--saveMACSPileup` * `--skipDiffAnalysis` * `--skip*` for skipping QC steps +* Update template to tools v1.7 ### `Fixed` From 2b9e42f749ab84e24e09cee29afb5ca78edb452c Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 10:20:11 +0100 Subject: [PATCH 068/149] Bump nextflow version --- .travis.yml | 2 +- README.md | 2 +- nextflow.config | 2 +- 3 files changed, 3 insertions(+), 3 deletions(-) diff --git a/.travis.yml b/.travis.yml index 9409abbe..f60b6d5b 100755 --- a/.travis.yml +++ b/.travis.yml @@ -30,7 +30,7 @@ install: - sudo apt-get install npm && npm install -g markdownlint-cli env: - - NXF_VER='0.32.0' # Specify a minimum NF version that should be tested and work + - NXF_VER='19.04.0' # Specify a minimum NF version that should be tested and work - NXF_VER='' # Plus: get the latest NF version and check that it works script: diff --git a/README.md b/README.md index 443cb2b5..912aa42c 100644 --- a/README.md +++ b/README.md @@ -1,7 +1,7 @@ # ![nfcore/atacseq](docs/images/nf-core-atacseq_logo.png) [![Build Status](https://travis-ci.org/nf-core/atacseq.svg?branch=master)](https://travis-ci.org/nf-core/atacseq) -[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A50.32.0-brightgreen.svg)](https://www.nextflow.io/) +[![Nextflow](https://img.shields.io/badge/nextflow-%E2%89%A519.04.0-brightgreen.svg)](https://www.nextflow.io/) [![install with bioconda](https://img.shields.io/badge/install%20with-bioconda-brightgreen.svg)](http://bioconda.github.io/) [![Docker](https://img.shields.io/docker/automated/nfcore/atacseq.svg)](https://hub.docker.com/r/nfcore/atacseq) diff --git a/nextflow.config b/nextflow.config index 0de69181..24baa462 100755 --- a/nextflow.config +++ b/nextflow.config @@ -147,7 +147,7 @@ manifest { homePage = 'https://github.com/nf-core/atacseq' description = 'ATACSeq peak-calling and differential analysis pipeline.' mainScript = 'main.nf' - nextflowVersion = '>=0.32.0' + nextflowVersion = '>=19.04.0' version = '1.0.1dev' } From 27ba903bbda651737fb1f3da2cb749b3af8152bd Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 10:26:13 +0100 Subject: [PATCH 069/149] Update CHANGELOG --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index df6c88a3..8fdad49b 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -24,7 +24,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * `--saveMACSPileup` * `--skipDiffAnalysis` * `--skip*` for skipping QC steps -* Update template to tools v1.7 +* Update template to tools `1.7` +* Bump Nextflow version to `19.04.0` ### `Fixed` From a79a050becc34721f6fc73eec26acc5903610b53 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 13:04:21 +0100 Subject: [PATCH 070/149] Update blacklist staging --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 80a73252..ee3c8611 100644 --- a/main.nf +++ b/main.nf @@ -174,7 +174,7 @@ if (params.design) { ch_design = file(params.design, checkIfExists: true) } e if (params.gtf) { ch_gtf = file(params.gtf, checkIfExists: true) } else { exit 1, "GTF annotation file not specified!" } if (params.gene_bed) { ch_gene_bed = file(params.gene_bed, checkIfExists: true) } if (params.tss_bed) { ch_tss_bed = file(params.tss_bed, checkIfExists: true) } -if (params.blacklist) { ch_blacklist = file(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } +if (params.blacklist) { ch_blacklist = Channel.fromPath(params.blacklist, checkIfExists: true) } else { ch_blacklist = Channel.empty() } if (params.fasta) { lastPath = params.fasta.lastIndexOf(File.separator) From 1809200230b419221d8de499c57c8bb74fb72ef6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 14:05:29 +0100 Subject: [PATCH 071/149] Update params --- main.nf | 24 ++++++++++++++---------- 1 file changed, 14 insertions(+), 10 deletions(-) diff --git a/main.nf b/main.nf index ee3c8611..9b7508a1 100644 --- a/main.nf +++ b/main.nf @@ -493,14 +493,14 @@ process FastQC { if (params.singleEnd) { """ [ ! -f ${name}.fastq.gz ] && ln -s $reads ${name}.fastq.gz - fastqc -q ${name}.fastq.gz + fastqc -q -t $task.cpus ${name}.fastq.gz """ } else { """ [ ! -f ${name}_1.fastq.gz ] && ln -s ${reads[0]} ${name}_1.fastq.gz [ ! -f ${name}_2.fastq.gz ] && ln -s ${reads[1]} ${name}_2.fastq.gz - fastqc -q ${name}_1.fastq.gz - fastqc -q ${name}_2.fastq.gz + fastqc -q -t $task.cpus ${name}_1.fastq.gz + fastqc -q -t $task.cpus ${name}_2.fastq.gz """ } } @@ -979,7 +979,7 @@ process MergeLibraryPlotProfile { --afterRegionStartLength 3000 \\ --skipZeros \\ --smartLabels \\ - -p $task.cpus + --numberOfProcessors $task.cpus plotProfile --matrixFile ${prefix}.computeMatrix.mat.gz \\ --outFileName ${prefix}.plotProfile.pdf \\ @@ -1017,8 +1017,8 @@ process MergeLibraryPlotFingerprint { --outRawCounts ${prefix}.plotFingerprint.raw.txt \\ --outQualityMetrics ${prefix}.plotFingerprint.qcmetrics.txt \\ --skipZeros \\ - --numberOfProcessors ${task.cpus} \\ - --numberOfSamples ${params.fingerprint_bins} + --numberOfProcessors $task.cpus \\ + --numberOfSamples $params.fingerprint_bins """ } @@ -1071,8 +1071,8 @@ process MergeLibraryMACSCallPeak { -t ${bam[0]} \\ $broad \\ -f $format \\ - -g ${params.macs_gsize} \\ - -n ${prefix} \\ + -g $params.macs_gsize \\ + -n $prefix \\ $pileup \\ --keep-dup all \\ --nomodel @@ -1114,6 +1114,7 @@ process MergeLibraryAnnotatePeaks { $fasta \\ -gid \\ -gtf $gtf \\ + -cpu $task.cpus \\ > ${prefix}_peaks.annotatePeaks.txt """ } @@ -1239,6 +1240,7 @@ process MergeLibraryConsensusPeakSetAnnotate { $fasta \\ -gid \\ -gtf $gtf \\ + -cpu $task.cpus \\ > ${prefix}.annotatePeaks.txt cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt @@ -1541,8 +1543,8 @@ process MergeReplicateMACSCallPeak { -t ${bam[0]} \\ $broad \\ -f $format \\ - -g ${params.macs_gsize} \\ - -n ${prefix} \\ + -g $params.macs_gsize \\ + -n $prefix \\ $pileup \\ --keep-dup all \\ --nomodel @@ -1584,6 +1586,7 @@ process MergeReplicateAnnotatePeaks { $fasta \\ -gid \\ -gtf $gtf \\ + -cpu $task.cpus \\ > ${prefix}_peaks.annotatePeaks.txt """ } @@ -1708,6 +1711,7 @@ process MergeReplicateConsensusPeakSetAnnotate { $fasta \\ -gid \\ -gtf $gtf \\ + -cpu $task.cpus \\ > ${prefix}.annotatePeaks.txt cut -f2- ${prefix}.annotatePeaks.txt | awk 'NR==1; NR > 1 {print \$0 | "sort -k1,1 -k2,2n"}' | cut -f6- > tmp.txt From 2ffa07a9559e3422d9946579d8796dfed0d847a6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 14:16:44 +0100 Subject: [PATCH 072/149] Missed 2M in header log info --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 9b7508a1..2328266b 100644 --- a/main.nf +++ b/main.nf @@ -282,7 +282,7 @@ if (params.email || params.email_on_fail) { summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize } log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join("\n") -log.info "\033[2m----------------------------------------------------\033[0m" +log.info "-\033[2m--------------------------------------------------\033[0m-" // Check the hostnames against configured profiles checkHostname() From 615ad2c9131b1ea55b3166ae4729bb649c48babc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 14:24:42 +0100 Subject: [PATCH 073/149] Fix MultiQC HOMER peak to feature plot --- assets/multiqc/mlib_peak_annotation_header.txt | 7 ++----- assets/multiqc/mrep_peak_annotation_header.txt | 7 ++----- 2 files changed, 4 insertions(+), 10 deletions(-) diff --git a/assets/multiqc/mlib_peak_annotation_header.txt b/assets/multiqc/mlib_peak_annotation_header.txt index 9bb837dc..3abac2f0 100644 --- a/assets/multiqc/mlib_peak_annotation_header.txt +++ b/assets/multiqc/mlib_peak_annotation_header.txt @@ -5,8 +5,5 @@ #plot_type: 'bargraph' #anchor: 'nfcore_atacseq-mlib_peak_annotation' #pconfig: -# title: 'Peak to feature %' -# ylab: 'Feature %' -# ymax: 100 -# ymin: 0 -# cpswitch_c_active: false +# title: 'Peak to feature proportion' +# ylab: 'Peak count' diff --git a/assets/multiqc/mrep_peak_annotation_header.txt b/assets/multiqc/mrep_peak_annotation_header.txt index 8e50d025..4e605e68 100644 --- a/assets/multiqc/mrep_peak_annotation_header.txt +++ b/assets/multiqc/mrep_peak_annotation_header.txt @@ -5,8 +5,5 @@ #plot_type: 'bargraph' #anchor: 'nfcore_atacseq-mrep_peak_annotation' #pconfig: -# title: 'Peak to feature %' -# ylab: 'Feature %' -# ymax: 100 -# ymin: 0 -# cpswitch_c_active: false +# title: 'Peak to feature proportion' +# ylab: 'Peak count' From 13e17595850a45438ced78d0296e14e65eebbbea Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 15:21:54 +0100 Subject: [PATCH 074/149] Update CHANGELOG --- CHANGELOG.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index 8fdad49b..40ee0efa 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -11,7 +11,6 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#35](https://github.com/nf-core/atacseq/issues/35) - Add deepTools plotFingerprint * [#46](https://github.com/nf-core/atacseq/issues/46) - Missing gene_bed path in igenomes config -* [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly * Merged in TEMPLATE branch for automated syncing * Renamed all channels to start with `ch_` prefix * Capitalised process names @@ -31,6 +30,8 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#41](https://github.com/nf-core/atacseq/issues/41) - Docs: Add example plot images * [#44](https://github.com/nf-core/atacseq/issues/44) - Output directory missing: macs2/consensus/deseq2 +* [#45](https://github.com/nf-core/atacseq/issues/45) - Wrong x-axis scale for the HOMER: Peak annotation Counts tab plot? +* [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly * Smaller logo for completion email * IGV uses relative instead of absolute paths From 4eb80ff3e8e64c40635a57e22e2012aa72221634 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 15:52:06 +0100 Subject: [PATCH 075/149] Fix levels in R scripts --- bin/plot_homer_annotatepeaks.r | 6 +++--- bin/plot_macs_qc.r | 2 +- 2 files changed, 4 insertions(+), 4 deletions(-) diff --git a/bin/plot_homer_annotatepeaks.r b/bin/plot_homer_annotatepeaks.r index ee537fce..9865357c 100755 --- a/bin/plot_homer_annotatepeaks.r +++ b/bin/plot_homer_annotatepeaks.r @@ -87,9 +87,9 @@ for (idx in 1:length(HomerFiles)) { plot.dist.dat <- rbind(plot.dist.dat,dist.melt) } -levels(plot.dat$name) <- sort(unique(as.character(plot.dat$name))) -levels(plot.dist.dat$variable) <- sort(unique(as.character(plot.dist.dat$variable))) -levels(plot.feature.dat$variable) <- sort(unique(as.character(plot.feature.dat$variable))) +plot.dat$name <- factor(plot.dat$name, levels=sort(unique(as.character(plot.dat$name)))) +plot.dist.dat$variable <- factor(plot.dist.dat$variable, levels=sort(unique(as.character(plot.dist.dat$variable)))) +plot.feature.dat$variable <- factor(plot.feature.dat$variable, levels=sort(unique(as.character(plot.feature.dat$variable)))) summary.dat <- dcast(plot.feature.dat, variable ~ feature, value.var="value") colnames(summary.dat)[1] <- "sample" diff --git a/bin/plot_macs_qc.r b/bin/plot_macs_qc.r index e43fcb8c..2360505f 100755 --- a/bin/plot_macs_qc.r +++ b/bin/plot_macs_qc.r @@ -81,7 +81,7 @@ for (idx in 1:length(PeakFiles)) { peaks.dat$name <- rep(sampleid,nrow(peaks.dat)) plot.dat <- rbind(plot.dat,peaks.dat) } -levels(plot.dat$name) <- sort(unique(as.character(plot.dat$name))) +plot.dat$name <- factor(plot.dat$name, levels=sort(unique(as.character(plot.dat$name)))) SummaryFile <- file.path(opt$outdir,paste(opt$outprefix,".summary.txt",sep="")) write.table(summary.dat,file=SummaryFile,quote=FALSE,sep="\t",row.names=FALSE,col.names=TRUE) From e9fdaf4fa0398b8e3fa2abf6b82d9c8ba2083bcc Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 15:55:54 +0100 Subject: [PATCH 076/149] Update CHANGELOG --- CHANGELOG.md | 1 + 1 file changed, 1 insertion(+) diff --git a/CHANGELOG.md b/CHANGELOG.md index 40ee0efa..ffacdd9a 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -32,6 +32,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * [#44](https://github.com/nf-core/atacseq/issues/44) - Output directory missing: macs2/consensus/deseq2 * [#45](https://github.com/nf-core/atacseq/issues/45) - Wrong x-axis scale for the HOMER: Peak annotation Counts tab plot? * [#46](https://github.com/nf-core/atacseq/issues/46) - Stage blacklist file in channel properly +* [#50](https://github.com/nf-core/atacseq/issues/50) - HOMER number of peaks does not correspond to found MACS2 peaks * Smaller logo for completion email * IGV uses relative instead of absolute paths From a2ebb478cc1f0facfde603de5fa52e02e1aadab7 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Fri, 11 Oct 2019 16:30:44 +0100 Subject: [PATCH 077/149] Fix MultiQC channel --- main.nf | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/main.nf b/main.nf index 2328266b..b0b31453 100644 --- a/main.nf +++ b/main.nf @@ -2004,7 +2004,7 @@ workflow.onComplete { def mqc_report = null try { if (workflow.success) { - mqc_report = multiqc_report.getVal() + mqc_report = ch_multiqc_report.getVal() if (mqc_report.getClass() == ArrayList) { log.warn "[nf-core/atacseq] Found multiple reports from process 'multiqc', will use only one" mqc_report = mqc_report[0] From abe587b75c87c9a64112f66954f027c385a69dcf Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 13:16:53 +0100 Subject: [PATCH 078/149] Change maxMultiqcEmailFileSize to max_multiqc_email_size --- docs/usage.md | 4 ++++ main.nf | 8 ++++---- nextflow.config | 2 +- 3 files changed, 9 insertions(+), 5 deletions(-) diff --git a/docs/usage.md b/docs/usage.md index fe069a78..eca4034a 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -55,6 +55,7 @@ * [`--outdir`](#--outdir) * [`--email`](#--email) * [`--email_on_fail`](#--email_on_fail) + * [`--max_multiqc_email_size`](#--max_multiqc_email_size) * [`-name`](#-name) * [`-resume`](#-resume) * [`-c`](#-c) @@ -410,6 +411,9 @@ Set this parameter to your e-mail address to get a summary e-mail with details o ### `--email_on_fail` This works exactly as with `--email`, except emails are only sent if the workflow is not successful. +### `--max_multiqc_email_size` +Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB). + ### `-name` Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic. diff --git a/main.nf b/main.nf index b0b31453..7aa8dd23 100644 --- a/main.nf +++ b/main.nf @@ -85,7 +85,7 @@ def helpMessage() { --outdir The output directory where the results will be saved --email Set this parameter to your e-mail address to get a summary e-mail with details of the run sent to you when the workflow exits --email_on_fail Same as --email, except only send mail if the workflow is not successful - --maxMultiqcEmailFileSize Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) + --max_multiqc_email_size Theshold size for MultiQC report to be attached in notification email. If file generated by pipeline exceeds the threshold, it will not be attached (Default: 25MB) -name Name for the pipeline run. If not specified, Nextflow will automatically generate a random mnemonic AWSBatch @@ -279,7 +279,7 @@ if (params.config_profile_url) summary['Config URL'] = params.co if (params.email || params.email_on_fail) { summary['E-mail Address'] = params.email summary['E-mail on failure'] = params.email_on_fail - summary['MultiQC Max Size'] = params.maxMultiqcEmailFileSize + summary['MultiQC Max Size'] = params.max_multiqc_email_size } log.info summary.collect { k,v -> "${k.padRight(21)}: $v" }.join("\n") log.info "-\033[2m--------------------------------------------------\033[0m-" @@ -1999,7 +1999,7 @@ workflow.onComplete { email_fields['summary']['Nextflow Build'] = workflow.nextflow.build email_fields['summary']['Nextflow Compile Timestamp'] = workflow.nextflow.timestamp - // TODO nf-core: If not using MultiQC, strip out this code (including params.maxMultiqcEmailFileSize) + // TODO nf-core: If not using MultiQC, strip out this code (including params.max_multiqc_email_size) // On success try attach the multiqc report def mqc_report = null try { @@ -2032,7 +2032,7 @@ workflow.onComplete { def email_html = html_template.toString() // Render the sendmail template - def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.maxMultiqcEmailFileSize.toBytes() ] + def smail_fields = [ email: email_address, subject: subject, email_txt: email_txt, email_html: email_html, baseDir: "$baseDir", mqcFile: mqc_report, mqcMaxSize: params.max_multiqc_email_size.toBytes() ] def sf = new File("$baseDir/assets/sendmail_template.txt") def sendmail_template = engine.createTemplate(sf).make(smail_fields) def sendmail_html = sendmail_template.toString() diff --git a/nextflow.config b/nextflow.config index 24baa462..7279d4bc 100755 --- a/nextflow.config +++ b/nextflow.config @@ -73,7 +73,7 @@ params { outdir = './results' igenomes_base = "./iGenomes" igenomesIgnore = false - maxMultiqcEmailFileSize = 25.MB + max_multiqc_email_size = 25.MB tracedir = "${params.outdir}/pipeline_info" email = false email_on_fail = false From 418dc89042b69a5901181a1a28cd4209cfcb1647 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 13:28:35 +0100 Subject: [PATCH 079/149] Change saveGenomeIndex to saveReference --- CHANGELOG.md | 1 + docs/output.md | 4 ++-- docs/usage.md | 8 ++++---- main.nf | 8 ++++---- nextflow.config | 2 +- 5 files changed, 12 insertions(+), 11 deletions(-) diff --git a/CHANGELOG.md b/CHANGELOG.md index ffacdd9a..e35548e9 100644 --- a/CHANGELOG.md +++ b/CHANGELOG.md @@ -23,6 +23,7 @@ and this project adheres to [Semantic Versioning](http://semver.org/spec/v2.0.0. * `--saveMACSPileup` * `--skipDiffAnalysis` * `--skip*` for skipping QC steps +* Change parameter `saveGenomeIndex` to `saveReference` * Update template to tools `1.7` * Bump Nextflow version to `19.04.0` diff --git a/docs/output.md b/docs/output.md index ef0f710e..f78591de 100755 --- a/docs/output.md +++ b/docs/output.md @@ -53,7 +53,7 @@ The initial QC and alignments are performed at the library-level e.g. if the sam [BWA](https://sourceforge.net/projects/bio-bwa/files/), [SAMtools](https://sourceforge.net/projects/samtools/files/samtools/) *Description*: - Adapter-trimmed reads are mapped to the reference assembly using BWA. A genome index is required to run BWA so if this isnt provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--saveGenomeIndex` parameter to save the indices for future pipeline runs, reducing processing times. + Adapter-trimmed reads are mapped to the reference assembly using BWA. A genome index is required to run BWA so if this isnt provided explicitly using the `--bwa_index` parameter then it will be created automatically from the genome fasta input. The index creation process can take a while for larger genomes so it is possible to use the `--saveReference` parameter to save the indices for future pipeline runs, reducing processing times. File names in the resulting directory (i.e. `bwa/library/`) will have the '`.Lb.`' (**L**i**b**rary) suffix. @@ -286,7 +286,7 @@ You can skip this portion of the analysis by specifying the `--skipMergeReplicat * `reference_genome/` A number of genome-specific files are generated by the pipeline in order to aid in the filtering of the data, and because they are required by standard tools such as BEDTools. These can be found in this directory along with the genome fasta file which is required by IGV. * `reference_genome/BWAIndex/` - If they dont exist already and if the `--saveGenomeIndex` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. + If they dont exist already and if the `--saveReference` parameter is provided then the alignment indices generated by the pipeline will be saved in this directory. This can be quite a time-consuming process so it permits their reuse for future runs of the pipeline or for other purposes. 2. **Pipeline information** diff --git a/docs/usage.md b/docs/usage.md index eca4034a..20b6e5c2 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -27,7 +27,7 @@ * [`--macs_gsize`](#--macs_gsize) * [`--blacklist`](#--blacklist) * [`--mito_name`](#--mito_name) - * [`--saveGenomeIndex`](#--savegenomeindex) + * [`--saveReference`](#--saveReference) * [`--igenomesIgnore`](#--igenomesignore) * [Adapter trimming](#adapter-trimming) * [`--skipTrimming`](#--skiptrimming) @@ -47,7 +47,7 @@ * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) - * [Custom resource requests](#custom-resource-requests) + * [CussaveReferencerequests](#custom-resource-requests) * [AWS Batch specific parameters](#aws-batch-specific-parameters) * [`--awsqueue`](#--awsqueue) * [`--awsregion`](#--awsregion) @@ -244,7 +244,7 @@ params { ``` ### `--fasta` -Full path to fasta file containing reference genome (*mandatory* if `--genome` is not specified). If you don't have a BWA index available this will be generated for you automatically. Combine with `--saveGenomeIndex` to save BWA index for future runs. +Full path to fasta file containing reference genome (*mandatory* if `--genome` is not specified). If you don't have a BWA index available this will be generated for you automatically. Combine with `--saveReference` to save BWA index for future runs. ```bash --fasta '[path to FASTA reference]' @@ -299,7 +299,7 @@ Name of mitochondrial chomosome in reference assembly. Reads aligning to this co --mito_name chrM ``` -### `--saveGenomeIndex` +### `--saveReference` If the BWA index is generated by the pipeline use this parameter to save it to your results folder. These can then be used for future pipeline runs, reducing processing times. ### `--igenomesIgnore` diff --git a/main.nf b/main.nf index 7aa8dd23..07e48114 100644 --- a/main.nf +++ b/main.nf @@ -47,7 +47,7 @@ def helpMessage() { --macs_gsize Effective genome size parameter required by MACS2. If using iGenomes config, values have only been provided when --genome is set as GRCh37, GRCm38, hg19, mm10, BDGP6 and WBcel235 --blacklist Path to blacklist regions (.BED format), used for filtering alignments --mito_name Name of Mitochondrial chomosome in genome fasta (e.g. chrM). Reads aligning to this contig are filtered out - --saveGenomeIndex If generated by the pipeline save the BWA index in the results directory + --saveReference If generated by the pipeline save the BWA index in the results directory Trimming --clip_r1 [int] Instructs Trim Galore to remove bp from the 5' end of read 1 (or single-end reads) (Default: 0) @@ -246,7 +246,7 @@ summary['Fingerprint Bins'] = params.fingerprint_bins if (params.keepMito) summary['Keep Mitochondrial'] = 'Yes' if (params.keepDups) summary['Keep Duplicates'] = 'Yes' if (params.keepMultiMap) summary['Keep Multi-mapped'] = 'Yes' -summary['Save Genome Index'] = params.saveGenomeIndex ? 'Yes' : 'No' +summary['Save Genome Index'] = params.saveReference ? 'Yes' : 'No' if (params.saveTrimmed) summary['Save Trimmed'] = 'Yes' if (params.saveAlignedIntermediates) summary['Save Intermeds'] = 'Yes' if (params.saveMACSPileup) summary['Save MACS2 Pileup'] = 'Yes' @@ -371,8 +371,8 @@ if (!params.bwa_index) { process BWAIndex { tag "$fasta" label 'process_high' - publishDir path: { params.saveGenomeIndex ? "${params.outdir}/reference_genome" : params.outdir }, - saveAs: { params.saveGenomeIndex ? it : null }, mode: 'copy' + publishDir path: { params.saveReference ? "${params.outdir}/reference_genome" : params.outdir }, + saveAs: { params.saveReference ? it : null }, mode: 'copy' input: file fasta from ch_fasta diff --git a/nextflow.config b/nextflow.config index 7279d4bc..f06eaf40 100755 --- a/nextflow.config +++ b/nextflow.config @@ -17,7 +17,7 @@ params { // Options: References genome = false tss_bed = false - saveGenomeIndex = false + saveReference = false // Options: Trimming clip_r1 = 0 From 55aadb35d0bf14045950e5c126f0d1b0ddde4ad1 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 14:29:24 +0100 Subject: [PATCH 080/149] Unify with chipseq --- main.nf | 64 +++++++++++++++++++++++++-------------------------------- 1 file changed, 28 insertions(+), 36 deletions(-) diff --git a/main.nf b/main.nf index 07e48114..0d76c623 100644 --- a/main.nf +++ b/main.nf @@ -9,14 +9,6 @@ ---------------------------------------------------------------------------------------- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// -/* -- -- */ -/* -- PARAMETERS -- */ -/* -- -- */ -/////////////////////////////////////////////////////////////////////////////// -/////////////////////////////////////////////////////////////////////////////// - def helpMessage() { log.info nfcoreHeader() log.info""" @@ -572,7 +564,7 @@ if (params.skipTrimming) { /* * STEP 3.1 - Align read 1 with bwa */ -process BWAmem { +process BWAMem { tag "$name" label 'process_high' @@ -585,9 +577,9 @@ process BWAmem { script: prefix = "${name}.Lb" - rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\'" if (params.seq_center) { - rg="\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" + rg = "\'@RG\\tID:${name}\\tSM:${name.split('_')[0..-2].join('_')}\\tPL:ILLUMINA\\tLB:${name}\\tPU:1\\tCN:${params.seq_center}\'" } """ bwa mem \\ @@ -869,7 +861,7 @@ process MergeLibraryPreseq { } /* - * STEP 5.2 Picard CollectMultipleMetrics after merging libraries + * STEP 5.2 Picard CollectMultipleMetrics after merging libraries and filtering */ process MergeLibraryMetrics { tag "$name" @@ -1068,14 +1060,14 @@ process MergeLibraryMACSCallPeak { pileup = params.saveMACSPileup ? "-B --SPMR" : "" """ macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel cat ${prefix}_peaks.${peaktype} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mlib_peak_count_header - > ${prefix}_peaks.count_mqc.tsv @@ -1191,7 +1183,7 @@ process MergeLibraryConsensusPeakSet { expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt macs2_merged_expand.py \\ ${prefix}.txt \\ @@ -1255,8 +1247,8 @@ process MergeLibraryConsensusPeakSetDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus/deseq2", mode: 'copy', saveAs: {filename -> - if (filename.endsWith(".igv.txt")) null - else filename + if (filename.endsWith(".igv.txt")) null + else filename } when: @@ -1540,14 +1532,14 @@ process MergeReplicateMACSCallPeak { pileup = params.saveMACSPileup ? "-B --SPMR" : "" """ macs2 callpeak \\ - -t ${bam[0]} \\ - $broad \\ - -f $format \\ - -g $params.macs_gsize \\ - -n $prefix \\ - $pileup \\ - --keep-dup all \\ - --nomodel + -t ${bam[0]} \\ + $broad \\ + -f $format \\ + -g $params.macs_gsize \\ + -n $prefix \\ + $pileup \\ + --keep-dup all \\ + --nomodel cat ${prefix}_peaks.${peaktype} | wc -l | awk -v OFS='\t' '{ print "${name}", \$1 }' | cat $mrep_peak_count_header - > ${prefix}_peaks.count_mqc.tsv @@ -1663,7 +1655,7 @@ process MergeReplicateConsensusPeakSet { expandparam = params.narrowPeak ? "--is_narrow_peak" : "" """ sort -k1,1 -k2,2n ${peaks.collect{it.toString()}.sort().join(' ')} \\ - | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt + | mergeBed -c $mergecols -o $collapsecols > ${prefix}.txt macs2_merged_expand.py \\ ${prefix}.txt \\ @@ -1835,7 +1827,7 @@ process get_software_versions { } output: - file 'software_versions_mqc.yaml' into software_versions_mqc + file 'software_versions_mqc.yaml' into ch_software_versions_mqc file "software_versions.csv" script: @@ -1892,7 +1884,7 @@ process MultiQC { input: file multiqc_config from ch_multiqc_config - file ('software_versions/*') from software_versions_mqc.collect() + file ('software_versions/*') from ch_software_versions_mqc.collect() file ('workflow_summary/*') from create_workflow_summary(summary) file ('fastqc/*') from ch_fastqc_reports_mqc.collect().ifEmpty([]) @@ -2040,7 +2032,7 @@ workflow.onComplete { // Send the HTML e-mail if (email_address) { try { - if ( params.plaintext_email ){ throw GroovyException('Send plaintext e-mail, not HTML') } + if (params.plaintext_email) { throw GroovyException('Send plaintext e-mail, not HTML') } // Try to send HTML e-mail using sendmail [ 'sendmail', '-t' ].execute() << sendmail_html log.info "[nf-core/atacseq] Sent summary e-mail to $email_address (sendmail)" @@ -2089,7 +2081,7 @@ workflow.onComplete { /////////////////////////////////////////////////////////////////////////////// /////////////////////////////////////////////////////////////////////////////// -def nfcoreHeader(){ +def nfcoreHeader() { // Log colors ANSI codes c_reset = params.monochrome_logs ? '' : "\033[0m"; c_dim = params.monochrome_logs ? '' : "\033[2m"; From a442143fd880edce01d46bd2d28226192d3b63eb Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 14:52:13 +0100 Subject: [PATCH 081/149] Adjust spacing --- main.nf | 158 +++++++++++++++++++++++++++++--------------------------- 1 file changed, 81 insertions(+), 77 deletions(-) diff --git a/main.nf b/main.nf index 0d76c623..939bc9f6 100644 --- a/main.nf +++ b/main.nf @@ -469,7 +469,9 @@ process FastQC { tag "$name" label 'process_medium' publishDir "${params.outdir}/fastqc", mode: 'copy', - saveAs: {filename -> filename.endsWith(".zip") ? "zips/$filename" : "$filename"} + saveAs: { filename -> + filename.endsWith(".zip") ? "zips/$filename" : "$filename" + } when: !params.skipFastQC @@ -517,12 +519,12 @@ if (params.skipTrimming) { tag "$name" label 'process_long' publishDir "${params.outdir}/trim_galore", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".html")) "fastqc/$filename" - else if (filename.endsWith(".zip")) "fastqc/zips/$filename" - else if (filename.endsWith("trimming_report.txt")) "logs/$filename" - else params.saveTrimmed ? filename : null - } + saveAs: { filename -> + if (filename.endsWith(".html")) "fastqc/$filename" + else if (filename.endsWith(".zip")) "fastqc/zips/$filename" + else if (filename.endsWith("trimming_report.txt")) "logs/$filename" + else params.saveTrimmed ? filename : null + } input: set val(name), file(reads) from ch_raw_reads_trimgalore @@ -601,11 +603,12 @@ process SortBAM { if (params.saveAlignedIntermediates) { publishDir path: "${params.outdir}/bwa/library", mode: 'copy', saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else filename } - } + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else filename + } + } input: set val(name), file(bam) from ch_bwa_bam @@ -646,12 +649,12 @@ process MergeLibraryBAM { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary", mode: 'copy', saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else params.saveAlignedIntermediates ? filename : null - } + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else params.saveAlignedIntermediates ? filename : null + } input: set val(name), file(bams) from ch_sort_bam_merge @@ -723,14 +726,15 @@ process MergeLibraryBAMFilter { label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', saveAs: { filename -> - if (params.singleEnd || params.saveAlignedIntermediates) { - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null } - } + if (params.singleEnd || params.saveAlignedIntermediates) { + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } + } input: set val(name), file(bam) from ch_mlib_bam_filter @@ -790,13 +794,13 @@ if (params.singleEnd) { label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', saveAs: { filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".sorted.bam")) filename - else if (filename.endsWith(".sorted.bam.bai")) filename - else null - } + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".sorted.bam")) filename + else if (filename.endsWith(".sorted.bam.bai")) filename + else null + } input: set val(name), file(bam) from ch_mlib_filter_bam @@ -868,10 +872,10 @@ process MergeLibraryMetrics { label 'process_medium' publishDir path: "${params.outdir}/bwa/mergedLibrary", mode: 'copy', saveAs: { filename -> - if (filename.endsWith("_metrics")) "picard_metrics/$filename" - else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" - else null - } + if (filename.endsWith("_metrics")) "picard_metrics/$filename" + else if (filename.endsWith(".pdf")) "picard_metrics/pdf/$filename" + else null + } when: !params.skipPicardMetrics @@ -909,10 +913,10 @@ process MergeLibraryBigWig { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/bigwig", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null } input: @@ -1029,10 +1033,10 @@ process MergeLibraryMACSCallPeak { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename } when: @@ -1156,9 +1160,9 @@ process MergeLibraryPeakQC { process MergeLibraryConsensusPeakSet { label 'process_long' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".igv.txt")) null - else filename + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename } when: @@ -1246,10 +1250,10 @@ process MergeLibraryConsensusPeakSetAnnotate { process MergeLibraryConsensusPeakSetDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedLibrary/macs/${peaktype}/consensus/deseq2", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } when: params.macs_gsize && !params.skipDiffAnalysis && multipleGroups && replicatesExist @@ -1379,12 +1383,12 @@ process MergeReplicateBAM { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".flagstat")) "samtools_stats/$filename" - else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" - else if (filename.endsWith(".stats")) "samtools_stats/$filename" - else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" - else filename + saveAs: { filename -> + if (filename.endsWith(".flagstat")) "samtools_stats/$filename" + else if (filename.endsWith(".idxstats")) "samtools_stats/$filename" + else if (filename.endsWith(".stats")) "samtools_stats/$filename" + else if (filename.endsWith(".metrics.txt")) "picard_metrics/$filename" + else filename } input: @@ -1462,10 +1466,10 @@ process MergeReplicateBigWig { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/bigwig", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith("scale_factor.txt")) "scale/$filename" - else if (filename.endsWith(".bigWig")) "$filename" - else null + saveAs: { filename -> + if (filename.endsWith("scale_factor.txt")) "scale/$filename" + else if (filename.endsWith(".bigWig")) "$filename" + else null } when: @@ -1502,10 +1506,10 @@ process MergeReplicateMACSCallPeak { tag "$name" label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".tsv")) "qc/$filename" - else if (filename.endsWith(".igv.txt")) null - else filename + saveAs: { filename -> + if (filename.endsWith(".tsv")) "qc/$filename" + else if (filename.endsWith(".igv.txt")) null + else filename } when: @@ -1628,9 +1632,9 @@ process MergeReplicatePeakQC { process MergeReplicateConsensusPeakSet { label 'process_long' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".igv.txt")) null - else filename + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename } when: @@ -1717,10 +1721,10 @@ process MergeReplicateConsensusPeakSetAnnotate { process MergeReplicateConsensusPeakSetDESeq { label 'process_medium' publishDir "${params.outdir}/bwa/mergedReplicate/macs/${peaktype}/consensus/deseq2", mode: 'copy', - saveAs: {filename -> - if (filename.endsWith(".igv.txt")) null - else filename - } + saveAs: { filename -> + if (filename.endsWith(".igv.txt")) null + else filename + } when: !params.skipMergeReplicates && !params.skipDiffAnalysis && replicatesExist && params.macs_gsize && multipleGroups @@ -1821,10 +1825,10 @@ process IGV { */ process get_software_versions { publishDir "${params.outdir}/pipeline_info", mode: 'copy', - saveAs: {filename -> - if (filename.indexOf(".csv") > 0) filename - else null - } + saveAs: { filename -> + if (filename.indexOf(".csv") > 0) filename + else null + } output: file 'software_versions_mqc.yaml' into ch_software_versions_mqc From 91868c5ca7f98bd664fb9d84c359176f20d8f550 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 15:05:48 +0100 Subject: [PATCH 082/149] Fix bug --- docs/usage.md | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/docs/usage.md b/docs/usage.md index 20b6e5c2..87b8cc2b 100755 --- a/docs/usage.md +++ b/docs/usage.md @@ -47,7 +47,7 @@ * [Skipping QC steps](#skipping-qc-steps) * [Job resources](#job-resources) * [Automatic resubmission](#automatic-resubmission) - * [CussaveReferencerequests](#custom-resource-requests) + * [Custom resource requests](#custom-resource-requests) * [AWS Batch specific parameters](#aws-batch-specific-parameters) * [`--awsqueue`](#--awsqueue) * [`--awsregion`](#--awsregion) From 93abc90c5887c2194d2375289804fa28f58fd5e6 Mon Sep 17 00:00:00 2001 From: drpatelh Date: Tue, 15 Oct 2019 15:39:09 +0100 Subject: [PATCH 083/149] Update images in output.md --- docs/images/igv_screenshot.png | Bin 308981 -> 0 bytes ..._annotatePeaks_feature_percentage_plot.png | Bin 80757 -> 0 bytes docs/images/mqc_cutadapt_plot.png | Bin 40209 -> 104682 bytes docs/images/mqc_deseq2_pca_plot.png | Bin 31816 -> 61615 bytes .../mqc_deseq2_sample_similarity_plot.png | Bin 50065 -> 76247 bytes .../mqc_featureCounts_assignment_plot.png | Bin 26375 -> 76240 bytes docs/images/mqc_frip_score_plot.png | Bin 28209 -> 74548 bytes docs/images/mqc_macs2_peak_count_plot.png | Bin 28387 -> 72526 bytes docs/images/mqc_picard_deduplication_plot.png | Bin 29974 -> 76669 bytes docs/images/mqc_picard_insert_size_plot.png | Bin 84701 -> 116371 bytes docs/images/mqc_samtools_idxstats_plot.png | Bin 98673 -> 113974 bytes docs/images/r_deseq2_ma_plot.png | Bin 244920 -> 211428 bytes docs/images/r_deseq2_volcano_plot.png | Bin 57295 -> 212008 bytes docs/images/r_upsetr_intersect_plot.png | Bin 39757 -> 0 bytes 14 files changed, 0 insertions(+), 0 deletions(-) delete mode 100755 docs/images/igv_screenshot.png delete mode 100755 docs/images/mqc_annotatePeaks_feature_percentage_plot.png mode change 100644 => 100755 docs/images/mqc_cutadapt_plot.png mode change 100644 => 100755 docs/images/mqc_featureCounts_assignment_plot.png mode change 100644 => 100755 docs/images/mqc_frip_score_plot.png mode change 100644 => 100755 docs/images/mqc_macs2_peak_count_plot.png mode change 100644 => 100755 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