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Merge pull request #574 from nf-core/taps
Taps
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.github/workflows/linting.yml

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pre-commit:
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runs-on: ubuntu-latest
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steps:
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- uses: actions/checkout@11bd71901bbe5b1630ceea73d27597364c9af683 # v4
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- uses: actions/checkout@08c6903cd8c0fde910a37f88322edcfb5dd907a8 # v5
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- name: Set up Python 3.13
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uses: actions/setup-python@a26af69be951a213d495a4c3e4e4022e16d87065 # v5
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- name: Set up Python 3.14
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uses: actions/setup-python@e797f83bcb11b83ae66e0230d6156d7c80228e7c # v6
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with:
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python-version: "3.13"
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python-version: "3.14"
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- name: Install pre-commit
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run: pip install pre-commit
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runs-on: ubuntu-latest
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steps:
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- name: Check out pipeline code
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uses: actions/checkout@11bd71901bbe5b1630ceea73d27597364c9af683 # v4
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uses: actions/checkout@08c6903cd8c0fde910a37f88322edcfb5dd907a8 # v5
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- name: Install Nextflow
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uses: nf-core/setup-nextflow@v2
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- uses: actions/setup-python@a26af69be951a213d495a4c3e4e4022e16d87065 # v5
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- uses: actions/setup-python@e797f83bcb11b83ae66e0230d6156d7c80228e7c # v6
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with:
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python-version: "3.13"
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python-version: "3.14"
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architecture: "x64"
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- name: read .nf-core.yml
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- name: Upload linting log file artifact
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if: ${{ always() }}
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uses: actions/upload-artifact@ea165f8d65b6e75b540449e92b4886f43607fa02 # v4
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uses: actions/upload-artifact@330a01c490aca151604b8cf639adc76d48f6c5d4 # v5
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with:
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name: linting-logs
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path: |

.github/workflows/linting_comment.yml

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run: echo "pr_number=$(cat linting-logs/PR_number.txt)" >> $GITHUB_OUTPUT
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- name: Post PR comment
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uses: marocchino/sticky-pull-request-comment@52423e01640425a022ef5fd42c6fb5f633a02728 # v2
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uses: marocchino/sticky-pull-request-comment@773744901bac0e8cbb5a0dc842800d45e9b2b405 # v2
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with:
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GITHUB_TOKEN: ${{ secrets.GITHUB_TOKEN }}
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number: ${{ steps.pr_number.outputs.pr_number }}

.prettierignore

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testing*
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*.pyc
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bin/
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.nf-test/
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ro-crate-metadata.json
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*.nf.test.snap
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modules/nf-core/
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subworkflows/nf-core/

CITATIONS.md

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> Felix Krueger, Simon R. Andrews, Bismark: a flexible aligner and methylation caller for Bisulfite-Seq applications, Bioinformatics, Volume 27, Issue 11, 1 June 2011, Pages 1571–1572, doi: [10.1093/bioinformatics/btr167](https://doi.org/10.1093/bioinformatics/btr167)
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- [BWA-MEM](https://arxiv.org/abs/1303.3997v2)
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> Li H: Aligning sequence reads, clone sequences and assembly contigs with BWA-MEM. arXiv 2013. doi: 10.48550/arXiv.1303.3997
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- [bwa-meth](https://arxiv.org/abs/1401.1129)
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> Pedersen, Brent S. and Eyring, Kenneth and De, Subhajyoti and Yang, Ivana V. and Schwartz, David A. Fast and accurate alignment of long bisulfite-seq reads, arXiv:1401.1129, doi: [10.48550/arXiv.1401.1129](https://doi.org/10.48550/arXiv.1401.1129)
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> Daley, T., Smith, A. Predicting the molecular complexity of sequencing libraries. Nat Methods 10, 325–327 (2013). doi: [10.1038/nmeth.2375](https://doi.org/10.1038/nmeth.2375)
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- [rastair](https://bitbucket.org/bsblabludwig/rastair/src/master/)
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- [Samtools](https://doi.org/10.1093/gigascience/giab008)
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> Petr Danecek, James K Bonfield, Jennifer Liddle, John Marshall, Valeriu Ohan, Martin O Pollard, Andrew Whitwham, Thomas Keane, Shane A McCarthy, Robert M Davies, Heng Li, Twelve years of SAMtools and BCFtools, GigaScience, Volume 10, Issue 2, February 2021, giab008, doi: [10.1093/gigascience/giab008](https://doi.org/10.1093/gigascience/giab008)

README.md

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**nf-core/methylseq** is a bioinformatics analysis pipeline used for Methylation (Bisulfite) sequencing data. It pre-processes raw data from FastQ inputs, aligns the reads and performs extensive quality-control on the results.
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![nf-core/methylseq metro map](docs/images/4.0.0_metromap.png)
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![nf-core/methylseq metro map](docs/images/taps_metromap.png)
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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 uses Docker / Singularity / Podman / Charliecloud / Apptainer containers making installation trivial and results highly reproducible.
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## Pipeline Summary
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The pipeline allows you to choose between running either [Bismark](https://github.com/FelixKrueger/Bismark) or [bwa-meth](https://github.com/brentp/bwa-meth) / [MethylDackel](https://github.com/dpryan79/methyldackel).
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The pipeline allows you to choose between running either [Bismark](https://github.com/FelixKrueger/Bismark), [bwa-meth](https://github.com/brentp/bwa-meth) / [MethylDackel](https://github.com/dpryan79/methyldackel) or [BWA-Mem](https://github.com/lh3/bwa) plus [rastair](https://bitbucket.org/bsblabludwig/rastair/src/master/) for for TAPS data processing. rastair can also be used with bwa-meth aligned reads by setting the aligner to `--aligner bwameth` and adding the flag `--taps`.
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Choose between workflows by using `--aligner bismark` (default, uses bowtie2 for alignment), `--aligner bismark_hisat` or `--aligner bwameth`. For higher performance, the pipeline can leverage the [Parabricks implementation of bwa-meth (fq2bammeth)](https://docs.nvidia.com/clara/parabricks/latest/documentation/tooldocs/man_fq2bam_meth.html), which implements the baseline tool `bwa-meth` in a performant method using fq2bam (BWA-MEM + GATK) as a backend for processing on GPU. To use this option, include the `gpu` profile along with `--aligner bwameth`.
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Choose between workflows by using `--aligner bismark` (default, uses bowtie2 for alignment), `--aligner bismark_hisat`, `--aligner bwameth` or `--aligner bwamem`. For higher performance, the pipeline can leverage the [Parabricks implementation of bwa-meth (fq2bammeth)](https://docs.nvidia.com/clara/parabricks/latest/documentation/tooldocs/man_fq2bam_meth.html) and the [Parabricks implementation of bwa-mem (fq2bammemh)](https://docs.nvidia.com/clara/parabricks/latest/documentation/tooldocs/man_fq2bam.html), which implement the baseline tools `bwa-meth` and `bwa-mem`. To use this option, include the `gpu` profile along with `--aligner bwameth` or `--aligner bwamem`.
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Note: For faster CPU runs with BWA-Meth, enable the BWA-MEM2 algorithm using `--use_mem2`. The GPU pathway (Parabricks) requires `-profile gpu` and a container runtime (Docker, Singularity, or Podman); Conda/Mamba are not supported for the GPU module.
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| Step | Bismark workflow | bwa-meth workflow |
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| -------------------------------------------- | ------------------------ | --------------------- |
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| Generate Reference Genome Index _(optional)_ | Bismark | bwa-meth |
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| Merge re-sequenced FastQ files | cat | cat |
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| Raw data QC | FastQC | FastQC |
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| Adapter sequence trimming | Trim Galore! | Trim Galore! |
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| Align Reads | Bismark (bowtie2/hisat2) | bwa-meth |
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| Deduplicate Alignments | Bismark | Picard MarkDuplicates |
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| Extract methylation calls | Bismark | MethylDackel |
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| Sample report | Bismark | - |
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| Summary Report | Bismark | - |
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| Alignment QC | Qualimap _(optional)_ | Qualimap _(optional)_ |
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| Sample complexity | Preseq _(optional)_ | Preseq _(optional)_ |
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| Project Report | MultiQC | MultiQC |
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| Step | Bismark workflow | bwa-meth workflow | bwa-mem + TAPS workflow |
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| -------------------------------------------- | ------------------------ | --------------------- | -------------------------- |
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| Generate Reference Genome Index _(optional)_ | Bismark | bwa-meth | bwa index |
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| Merge re-sequenced FastQ files | cat | cat | cat |
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| Raw data QC | FastQC | FastQC | FastQC |
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| Adapter sequence trimming | Trim Galore! | Trim Galore! | Trim Galore! |
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| Align Reads | Bismark (bowtie2/hisat2) | bwa-meth | bwa mem |
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| Deduplicate Alignments | Bismark | Picard MarkDuplicates | Picard MarkDuplicates |
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| Extract methylation calls | Bismark | MethylDackel | TAPS subworkflow (rastair) |
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| Sample report | Bismark | - | - |
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| Summary Report | Bismark | - | - |
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| Alignment QC | Qualimap _(optional)_ | Qualimap _(optional)_ | Qualimap _(optional)_ |
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| Sample complexity | Preseq _(optional)_ | Preseq _(optional)_ | Preseq _(optional)_ |
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| Project Report | MultiQC | MultiQC | MultiQC |
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Optional targeted sequencing analysis is available via `--run_targeted_sequencing` and `--target_regions_file`; see the [usage documentation](https://nf-co.re/methylseq/usage) for details.
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conf/base.config

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withName: PARABRICKS_FQ2BAMMETH {
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memory = { 100.GB * task.attempt }
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}
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withName: PARABRICKS_FQ2BAM {
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memory = { 100.GB * task.attempt }
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}
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}
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process {
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withName: GATK4_REMOVEDUPLICATES {
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ext.args = "--REMOVE_DUPLICATES true --TAG_DUPLICATE_SET_MEMBERS true"
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}
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}
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process {
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withName: PICARD_ADDORREPLACEREADGROUPS {
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ext.args = "--RGID 1 --RGLB lib1 --RGPL illumina --RGPU unit1 --RGSM sample1"
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}
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}
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process {
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withName: PICARD_REMOVEDUPLICATES {
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ext.args = "--ASSUME_SORTED true --REMOVE_DUPLICATES true --VALIDATION_STRINGENCY LENIENT --PROGRAM_RECORD_ID 'null' --TMP_DIR tmp"
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ext.prefix = { "${meta.id}.dedup.sorted" }
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publishDir = [
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[
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path: { "${params.outdir}/${params.aligner}/deduplicated/picard_metrics" },
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pattern: "*.metrics.txt",
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mode: params.publish_dir_mode
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],
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[
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path: { "${params.outdir}/${params.aligner}/deduplicated" },
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pattern: "*.bam",
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mode: params.publish_dir_mode
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]
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]
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}
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}

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