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.editorconfig

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[*.{yml,yaml}]
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indent_size = 2
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[*.json]
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insert_final_newline = unset
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# These files are edited and tested upstream in nf-core/modules
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[/modules/nf-core/**]
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charset = unset

docs/usage.md

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* The `--gtf_group_features_type` parameter will automatically be set to `gene_type` as opposed to `gene_biotype`, respectively.
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* If you are running Salmon, the `--gencode` flag will also be passed to the index building step to overcome parsing issues resulting from the transcript IDs in GENCODE fasta files being separated by vertical pipes (`|`) instead of spaces (see [this issue](https://github.com/COMBINE-lab/salmon/issues/15)).
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## Samplesheet input
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You will need to create a samplesheet with information about the samples you would like to analyse before running the pipeline. Use this parameter to specify its location. It has to be a comma-separated file with 3 columns, and a header row as shown in the examples below.
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```console
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--input '[path to samplesheet file]'
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```
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### Multiple runs of the same sample
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The `sample` identifiers have to be the same when you have re-sequenced the same sample more than once e.g. to increase sequencing depth. The pipeline will concatenate the raw reads before performing any downstream analysis. Below is an example for the same sample sequenced across 3 lanes:
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```console
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sample,fastq_1,fastq_2
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CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
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CONTROL_REP1,AEG588A1_S1_L003_R1_001.fastq.gz,AEG588A1_S1_L003_R2_001.fastq.gz
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CONTROL_REP1,AEG588A1_S1_L004_R1_001.fastq.gz,AEG588A1_S1_L004_R2_001.fastq.gz
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```
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### Full samplesheet
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The pipeline will auto-detect whether a sample is single- or paired-end using the information provided in the samplesheet. The samplesheet can have as many columns as you desire, however, there is a strict requirement for the first 3 columns to match those defined in the table below.
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A final samplesheet file consisting of both single- and paired-end data may look something like the one below. This is for 6 samples, where `TREATMENT_REP3` has been sequenced twice.
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```console
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sample,fastq_1,fastq_2
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CONTROL_REP1,AEG588A1_S1_L002_R1_001.fastq.gz,AEG588A1_S1_L002_R2_001.fastq.gz
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CONTROL_REP2,AEG588A2_S2_L002_R1_001.fastq.gz,AEG588A2_S2_L002_R2_001.fastq.gz
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CONTROL_REP3,AEG588A3_S3_L002_R1_001.fastq.gz,AEG588A3_S3_L002_R2_001.fastq.gz
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TREATMENT_REP1,AEG588A4_S4_L003_R1_001.fastq.gz,
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TREATMENT_REP2,AEG588A5_S5_L003_R1_001.fastq.gz,
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TREATMENT_REP3,AEG588A6_S6_L003_R1_001.fastq.gz,
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TREATMENT_REP3,AEG588A6_S6_L004_R1_001.fastq.gz,
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```
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| Column | Description |
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|----------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
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| `sample` | Custom sample name. This entry will be identical for multiple sequencing libraries/runs from the same sample. Spaces in sample names are automatically converted to underscores (`_`). |
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| `fastq_1` | Full path to FastQ file for Illumina short reads 1. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". |
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| `fastq_2` | Full path to FastQ file for Illumina short reads 2. File has to be gzipped and have the extension ".fastq.gz" or ".fq.gz". |
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An [example samplesheet](../assets/samplesheet.csv) has been provided with the pipeline.
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## Running the pipeline
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The typical command for running the pipeline is as follows:

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