This workflow does not specify a "main" workflow file.
Workflow Type: Galaxy
ChIP-seq paired-end Workflow
Inputs dataset
- The workflow needs a single input which is a list of dataset pairs of fastqsanger.
Inputs values
- adapters sequences: this depends on the library preparation. If you don't know, use FastQC to determine if it is Truseq or Nextera.
- reference_genome: this field will be adapted to the genomes available for bowtie2.
- effective_genome_size: this is used by MACS2 and may be entered manually (indications are provided for heavily used genomes).
Processing
- The workflow will remove illumina adapters and low quality bases and filter out any pair with mate smaller than 15bp.
- The filtered reads are mapped with bowtie2 with default parameters.
- The BAM is filtered to keep only MAPQ30 and concordant pairs.
- The peaks are called with MACS2 which at the same time generates a coverage file.
- The coverage is converted to bigwig.
- A MultiQC is run to have an overview of the QC.
Warning
- The coverage output is not normalized.
- The filtered bam still has PCR duplicates which are removed by MACS2.
Contribution
@lldelisle wrote the workflow.
@nagoue updated the tools, made it work in usegalaxy.org, fixed the best practices and wrote the tests.
Inputs
ID | Name | Description | Type |
---|---|---|---|
PE fastq input | PE fastq input | Should be a paired collection with ChIPseq fastqs |
|
adapter_forward | adapter_forward | Please use: For R1: - For Nextera: CTGTCTCTTATACACATCTCCGAGCCCACGAGAC - For TrueSeq: GATCGGAAGAGCACACGTCTGAACTCCAGTCAC or AGATCGGAAGAGCACACGTCTGAACTCCAGTCAC |
|
adapter_reverse | adapter_reverse | Please use: For R2: - For Nextera: CTGTCTCTTATACACATCTGACGCTGCCGACGA - For TruSeq: GATCGGAAGAGCGTCGTGTAGGGAAAGAGTGT or AGATCGGAAGAGCGTCGTGTAGGGAAAGAGTGT |
|
effective_genome_size | effective_genome_size | Used by MACS2: H. sapiens: 2700000000, M. musculus: 1870000000, D. melanogaster: 120000000, C. elegans: 90000000 |
|
reference_genome | reference_genome | reference_genome |
|
Steps
ID | Name | Description |
---|---|---|
5 | Cutadapt (remove adapter + bad quality bases) | toolshed.g2.bx.psu.edu/repos/lparsons/cutadapt/cutadapt/4.0+galaxy0 |
6 | Bowtie2 map on reference | toolshed.g2.bx.psu.edu/repos/devteam/bowtie2/bowtie2/2.4.5+galaxy1 |
7 | filter MAPQ30 concordent pairs | toolshed.g2.bx.psu.edu/repos/devteam/samtool_filter2/samtool_filter2/1.8+galaxy1 |
8 | Call Peaks with MACS2 | toolshed.g2.bx.psu.edu/repos/iuc/macs2/macs2_callpeak/2.2.7.1+galaxy0 |
9 | summary of MACS2 | summary of MACS2 toolshed.g2.bx.psu.edu/repos/bgruening/text_processing/tp_grep_tool/1.1.1 |
10 | Bigwig from MACS2 | wig_to_bigWig |
11 | MultiQC | toolshed.g2.bx.psu.edu/repos/iuc/multiqc/multiqc/1.11+galaxy0 |
Outputs
ID | Name | Description | Type |
---|---|---|---|
mapping stats | mapping stats | n/a |
|
filtered BAM | filtered BAM | n/a |
|
MACS2 summits | MACS2 summits | n/a |
|
MACS2 peaks | MACS2 peaks | n/a |
|
MACS2 narrowPeak | MACS2 narrowPeak | n/a |
|
MACS2 report | MACS2 report | n/a |
|
coverage from MACS2 | coverage from MACS2 | n/a |
|
MultiQC on input dataset(s): Stats | MultiQC on input dataset(s): Stats | n/a |
|
MultiQC webpage | MultiQC webpage | n/a |
|
Version History
v0.1 (earliest) Created 15th Oct 2022 at 03:01 by WorkflowHub Bot
Updated to v0.1
Frozen
v0.1
3142fa5