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chipseq-se.cwl
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chipseq-se.cwl
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cwlVersion: v1.0
class: Workflow
requirements:
- class: SubworkflowFeatureRequirement
- class: ScatterFeatureRequirement
- class: StepInputExpressionRequirement
- class: MultipleInputFeatureRequirement
- class: InlineJavascriptRequirement
expressionLib:
- var get_root = function(basename) {
return basename.split('.').slice(0,1).join('.');
};
'sd:metadata':
- "../metadata/chipseq-header.cwl"
'sd:upstream':
genome_indices: "genome-indices.cwl"
control_file: "chipseq-se.cwl"
inputs:
indices_folder:
type: Directory
'sd:upstreamSource': "genome_indices/bowtie_indices"
label: "Genome indices"
doc: "Directory with the genome indices generated by Bowtie"
annotation_file:
type: File
'sd:upstreamSource': "genome_indices/annotation"
label: "Genome annotation file"
format: "http://edamontology.org/format_3475"
doc: "Genome annotation file in TSV format"
genome_size:
type: string
'sd:upstreamSource': "genome_indices/genome_size"
label: "Effective genome size"
doc: "The length of the mappable genome (hs, mm, ce, dm or number, for example 2.7e9)"
chrom_length:
type: File
'sd:upstreamSource': "genome_indices/chrom_length"
label: "Chromosome lengths file"
format: "http://edamontology.org/format_2330"
doc: "Chromosome lengths file in TSV format"
control_file:
type: File?
default: null
'sd:upstreamSource': "control_file/bambai_pair"
'sd:localLabel': true
label: "Control ChIP-Seq single-read experiment"
format: "http://edamontology.org/format_2572"
doc: "Indexed BAM file from the ChIP-Seq single-read experiment to be used as a control for MACS2 peak calling"
broad_peak:
type: boolean?
default: False
# 'sd:parent': "https://raw.githubusercontent.com/datirium/workflows/master/tags/antibody-dummy.cwl"
label: "Call broad peaks"
doc: "Make MACS2 call broad peaks by linking nearby highly enriched regions"
fastq_file:
type: File
label: "FASTQ file"
format: "http://edamontology.org/format_1930"
doc: "Single-read sequencing data in FASTQ format (fastq, fq, bzip2, gzip, zip)"
exp_fragment_size:
type: int?
default: 150
'sd:layout':
advanced: true
label: "Expected fragment size"
doc: "Expected fragment size for read extenstion towards 3' end if force_fragment_size was set to True or if calculated by MACS2 fragment size was less that 80 bp"
force_fragment_size:
type: boolean?
default: false
'sd:layout':
advanced: true
label: "Force peak calling with expected fragment size"
doc: "Make MACS2 don't build the shifting model and use expected fragment size for read extenstion towards 3' end"
clip_3p_end:
type: int?
default: 0
'sd:layout':
advanced: true
label: "Clip from 3' end"
doc: "Number of base pairs to clip from 3' end"
clip_5p_end:
type: int?
default: 0
'sd:layout':
advanced: true
label: "Clip from 5' end"
doc: "Number of base pairs to clip from 5' end"
remove_duplicates:
type: boolean?
default: false
'sd:layout':
advanced: true
label: "Remove PCR duplicates"
doc: "Remove PCR duplicates from sorted BAM file"
promoter_dist:
type: int?
default: 1000
'sd:layout':
advanced: true
label: "Max distance from gene TSS (in both direction) overlapping which the peak will be assigned to the promoter region"
doc: "Max distance from gene TSS (in both direction) overlapping which the peak will be assigned to the promoter region"
upstream_dist:
type: int?
default: 20000
'sd:layout':
advanced: true
label: "Max distance from the promoter (only in upstream direction) overlapping which the peak will be assigned to the upstream region"
doc: "Max distance from the promoter (only in upstream direction) overlapping which the peak will be assigned to the upstream region"
threads:
type: int?
default: 2
'sd:layout':
advanced: true
label: "Number of threads"
doc: "Number of threads for those steps that support multithreading"
outputs:
unaligned_fastq:
type:
- "null"
- File[]
format: "http://edamontology.org/format_1930"
label: "Unaligned FASTQ file(s)"
doc: "Unaligned FASTQ file(s)"
outputSource: bowtie_aligner/unaligned_fastq
multimapped_fastq:
type:
- "null"
- File[]
format: "http://edamontology.org/format_1930"
label: "Multimapped FASTQ file(s)"
doc: "Multimapped FASTQ file(s)"
outputSource: bowtie_aligner/multimapped_fastq
bigwig:
type: File
format: "http://edamontology.org/format_3006"
label: "Genome coverage"
doc: "Genome coverage in bigWig format"
outputSource: bam_to_bigwig/bigwig_file
'sd:visualPlugins':
- igvbrowser:
tab: 'IGV Genome Browser'
id: 'igvbrowser'
optional: true
type: 'wig'
name: "Genome Coverage"
height: 120
fastx_statistics:
type: File
label: "FASTQ quality statistics"
format: "http://edamontology.org/format_2330"
doc: "FASTQ quality statistics in TSV format"
outputSource: fastx_quality_stats/statistics_file
'sd:visualPlugins':
- line:
tab: 'QC Plots'
Title: 'Base Frequency Plot'
xAxisTitle: 'Nucleotide position'
yAxisTitle: 'Frequency'
colors: ["#b3de69", "#888888", "#fb8072", "#fdc381", "#99c0db"]
data: [$13, $14, $15, $16, $17]
- boxplot:
tab: 'QC Plots'
Title: 'Base Quality Plot'
xAxisTitle: 'Nucleotide position'
yAxisTitle: 'Quality score'
colors: ["#b3de69", "#888888", "#fb8072", "#fdc381", "#99c0db"]
data: [$11, $7, $8, $9, $12]
bowtie_log:
type: File
label: "Read alignment log"
format: "http://edamontology.org/format_2330"
doc: "Read alignment log file from Bowtie"
outputSource: bowtie_aligner/log_file
iaintersect_result:
type: File
label: "Gene annotated peaks"
format: "http://edamontology.org/format_3475"
doc: "MACS2 peak file annotated with nearby genes"
outputSource: island_intersect/result_file
'sd:visualPlugins':
- syncfusiongrid:
tab: 'Peak Calling'
Title: 'Peak Coordinates'
atdp_result:
type: File
label: "Average Tag Density Plot"
format: "http://edamontology.org/format_3475"
doc: "Average Tag Density Plot file in TSV format"
outputSource: average_tag_density/result_file
'sd:visualPlugins':
- scatter:
tab: 'QC Plots'
Title: 'Average Tag Density Plot'
xAxisTitle: 'Distance From TSS (bp)'
yAxisTitle: 'Average Tag Density (per bp)'
colors: ["#b3de69"]
height: 500
data: [$1, $2]
bambai_pair:
type: File
format: "http://edamontology.org/format_2572"
label: "Aligned reads"
doc: "Coordinate sorted BAM alignment and index BAI files"
outputSource: samtools_sort_index_after_rmdup/bam_bai_pair
'sd:visualPlugins':
- igvbrowser:
tab: 'IGV Genome Browser'
id: 'igvbrowser'
type: 'alignment'
format: 'bam'
name: "Nucleotide Sequence Alignments"
displayMode: "SQUISHED"
macs2_called_peaks:
type: File
label: "Called peaks"
format: "http://edamontology.org/format_3468"
doc: "Called peaks file with 1-based coordinates in XLS format"
outputSource: macs2_callpeak/peak_xls_file
macs2_narrow_peaks:
type: File?
label: "Narrow peaks"
format: "http://edamontology.org/format_3613"
doc: "Called peaks file in ENCODE narrow peak format"
outputSource: macs2_callpeak/narrow_peak_file
'sd:visualPlugins':
- igvbrowser:
tab: 'IGV Genome Browser'
id: 'igvbrowser'
type: 'annotation'
name: "Narrow peaks"
height: 120
macs2_broad_peaks:
type: File?
label: "Broad peaks"
format: "http://edamontology.org/format_3614"
doc: "Called peaks file in ENCODE broad peak format"
outputSource: macs2_callpeak/broad_peak_file
'sd:visualPlugins':
- igvbrowser:
tab: 'IGV Genome Browser'
id: 'igvbrowser'
type: 'annotation'
name: "Broad peaks"
height: 120
workflow_statistics_yaml:
type: File?
label: "YAML formatted combined log"
format: "http://edamontology.org/format_3750"
doc: "YAML formatted combined log"
outputSource: get_statistics/collected_statistics_yaml
workflow_statistics_markdown:
type: File?
label: "Markdown formatted combined log"
format: "http://edamontology.org/format_3835"
doc: "Markdown formatted combined log"
outputSource: get_statistics/collected_statistics_md
'sd:visualPlugins':
- markdownView:
tab: 'Overview'
workflow_statistics_tsv:
type: File
label: "Workflow execution statistics"
format: "http://edamontology.org/format_3475"
doc: "Overall workflow execution statistics from bowtie_aligner and samtools_rmdup steps"
outputSource: get_statistics/collected_statistics_tsv
'sd:visualPlugins':
- tableView:
vertical: true
tab: 'Overview'
'sd:preview':
'sd:visualPlugins':
- pie:
colors: ['#b3de69', '#99c0db', '#fb8072', '#fdc381']
data: [$2, $3, $4, $5]
bam_statistics_report:
type: File
label: "BAM statistics report (original)"
format: "http://edamontology.org/format_2330"
doc: "BAM statistics report (right after alignment and sorting)"
outputSource: get_bam_statistics/log_file
bam_statistics_report_after_filtering:
type: File
label: "BAM statistics report (after filtering)"
format: "http://edamontology.org/format_2330"
doc: "BAM statistics report (after all filters applied)"
outputSource: get_bam_statistics_after_filtering/log_file
preseq_estimates:
type: File?
label: "Expected Distinct Reads Count Plot"
format: "http://edamontology.org/format_3475"
doc: "Expected distinct reads count file from Preseq in TSV format"
outputSource: preseq/estimates_file
'sd:visualPlugins':
- scatter:
tab: 'QC Plots'
Title: 'Expected Distinct Reads Count Plot'
xAxisTitle: 'Total reads count'
yAxisTitle: 'Expected distinct reads count'
colors: ["#4b78a3"]
height: 500
data: [$1, $2]
estimated_fragment_size:
type: int
label: "Estimated fragment size"
doc: "Estimated fragment size for downstream analyses"
outputSource: macs2_callpeak/macs2_fragments_calculated
mapped_reads_number:
type: int
label: "Mapped reads number"
doc: "Mapped reads number for downstream analyses"
outputSource: get_statistics/mapped_reads
steps:
extract_fastq:
run: ../tools/extract-fastq.cwl
in:
compressed_file: fastq_file
out: [fastq_file]
fastx_quality_stats:
run: ../tools/fastx-quality-stats.cwl
in:
input_file: extract_fastq/fastq_file
out: [statistics_file]
bowtie_aligner:
run: ../tools/bowtie-alignreads.cwl
in:
upstream_filelist: extract_fastq/fastq_file
indices_folder: indices_folder
clip_3p_end: clip_3p_end
clip_5p_end: clip_5p_end
v:
default: 3
m:
default: 1
best:
default: true
strata:
default: true
sam:
default: true
unaligned_prefix:
default: "unaligned_reads"
multimapped_prefix:
default: "multimapped_reads"
threads: threads
q:
default: true
X:
default: 500
out:
- sam_file
- log_file
- unaligned_fastq
- multimapped_fastq
samtools_sort_index:
run: ../tools/samtools-sort-index.cwl
in:
sort_input: bowtie_aligner/sam_file
threads: threads
out: [bam_bai_pair]
preseq:
run: ../tools/preseq-lc-extrap.cwl
in:
bam_file: samtools_sort_index/bam_bai_pair
extrapolation:
default: 1000000000
out: [estimates_file]
samtools_rmdup:
run: ../tools/samtools-rmdup.cwl
in:
trigger: remove_duplicates
bam_file: samtools_sort_index/bam_bai_pair
single_end:
default: true
out:
- rmdup_output
- rmdup_log
samtools_sort_index_after_rmdup:
run: ../tools/samtools-sort-index.cwl
in:
trigger: remove_duplicates
sort_input: samtools_rmdup/rmdup_output
threads: threads
out: [bam_bai_pair]
macs2_callpeak:
run: ../tools/macs2-callpeak-biowardrobe-only.cwl
in:
treatment_file: samtools_sort_index_after_rmdup/bam_bai_pair
control_file: control_file
nolambda:
source: control_file
valueFrom: $(!self)
genome_size: genome_size
mfold:
default: "4 40"
verbose:
default: 3
nomodel: force_fragment_size
extsize: exp_fragment_size
bw: exp_fragment_size
broad: broad_peak
call_summits:
source: broad_peak
valueFrom: $(!self)
keep_dup:
default: auto
q_value:
default: 0.05
format_mode:
default: BAM
buffer_size:
default: 10000
out:
- peak_xls_file
- narrow_peak_file
- broad_peak_file
- macs2_fragments_calculated
bam_to_bigwig:
run: ../tools/bam-bedgraph-bigwig.cwl
in:
bam_file: samtools_sort_index_after_rmdup/bam_bai_pair
chrom_length_file: chrom_length
mapped_reads_number: get_statistics/mapped_reads
fragment_size: macs2_callpeak/macs2_fragments_calculated
out: [bigwig_file]
get_bam_statistics:
run: ../tools/samtools-stats.cwl
in:
bambai_pair: samtools_sort_index/bam_bai_pair
output_filename:
source: samtools_sort_index/bam_bai_pair
valueFrom: $(get_root(self.basename)+"_bam_statistics_report.txt")
out: [log_file]
get_bam_statistics_after_filtering:
run: ../tools/samtools-stats.cwl
in:
bambai_pair: samtools_sort_index_after_rmdup/bam_bai_pair
output_filename:
source: samtools_sort_index_after_rmdup/bam_bai_pair
valueFrom: $(get_root(self.basename)+"_bam_statistics_report_after_filtering.txt")
out: [log_file]
get_statistics:
run: ../tools/collect-statistics-chip-seq.cwl
in:
bowtie_alignment_report: bowtie_aligner/log_file
bam_statistics_report: get_bam_statistics/log_file
bam_statistics_after_filtering_report: get_bam_statistics_after_filtering/log_file
macs2_called_peaks: macs2_callpeak/peak_xls_file
preseq_results: preseq/estimates_file
out: [collected_statistics_yaml, collected_statistics_tsv, mapped_reads, collected_statistics_md]
island_intersect:
run: ../tools/iaintersect.cwl
in:
input_filename: macs2_callpeak/peak_xls_file
annotation_filename: annotation_file
promoter_bp: promoter_dist
upstream_bp: upstream_dist
out: [result_file]
average_tag_density:
run: ../tools/atdp.cwl
in:
input_file: samtools_sort_index_after_rmdup/bam_bai_pair
annotation_filename: annotation_file
fragmentsize_bp: macs2_callpeak/macs2_fragments_calculated
avd_window_bp:
default: 5000
avd_smooth_bp:
default: 50
ignore_chr:
default: chrM
double_chr:
default: "chrX chrY"
avd_heat_window_bp:
default: 200
mapped_reads: get_statistics/mapped_reads
out: [result_file]
$namespaces:
s: http://schema.org/
$schemas:
- https://github.com/schemaorg/schemaorg/raw/main/data/releases/11.01/schemaorg-current-http.rdf
label: "ChIP-Seq pipeline single-read"
s:name: "ChIP-Seq pipeline single-read"
s:alternateName: "ChIP-Seq basic analysis workflow for single-read data"
s:downloadUrl: https://raw.githubusercontent.com/datirium/workflows/master/workflows/chipseq-se.cwl
s:codeRepository: https://github.com/datirium/workflows
s:license: http://www.apache.org/licenses/LICENSE-2.0
s:isPartOf:
class: s:CreativeWork
s:name: Common Workflow Language
s:url: http://commonwl.org/
s:creator:
- class: s:Organization
s:legalName: "Cincinnati Children's Hospital Medical Center"
s:location:
- class: s:PostalAddress
s:addressCountry: "USA"
s:addressLocality: "Cincinnati"
s:addressRegion: "OH"
s:postalCode: "45229"
s:streetAddress: "3333 Burnet Ave"
s:telephone: "+1(513)636-4200"
s:logo: "https://www.cincinnatichildrens.org/-/media/cincinnati%20childrens/global%20shared/childrens-logo-new.png"
s:department:
- class: s:Organization
s:legalName: "Allergy and Immunology"
s:department:
- class: s:Organization
s:legalName: "Barski Research Lab"
s:member:
- class: s:Person
s:name: Michael Kotliar
s:email: mailto:michael.kotliar@cchmc.org
s:sameAs:
- id: http://orcid.org/0000-0002-6486-3898
# doc:
# $include: ../descriptions/chipseq-se.md
doc: |
# ChIP-Seq basic analysis workflow for single-read data
Reads are aligned to the reference genome with [Bowtie](http://bowtie-bio.sourceforge.net/index.shtml). Results are saved as coordinate sorted [BAM](http://samtools.github.io/hts-specs/SAMv1.pdf) alignment and index BAI files. Optionally, PCR duplicates can be removed. To obtain coverage in [bigWig](https://genome.ucsc.edu/goldenpath/help/bigWig.html) format, average fragment length is calculated by [MACS2](https://github.com/taoliu/MACS), and individual reads are extended to this length in the 3’ direction. Areas of enrichment identified by MACS2 are saved in ENCODE [narrow peak](http://genome.ucsc.edu/FAQ/FAQformat.html#format12) or [broad peak](https://genome.ucsc.edu/FAQ/FAQformat.html#format13) formats. Called peaks together with the nearest genes are saved in TSV format. In addition to basic statistics (number of total/mapped/multi-mapped/unmapped/duplicate reads), pipeline generates several quality control measures. Base frequency plots are used to estimate adapter contamination, a frequent occurrence in low-input ChIP-Seq experiments. Expected distinct reads count from [Preseq](http://smithlabresearch.org/software/preseq/) can be used to estimate read redundancy for a given sequencing depth. Average tag density profiles can be used to estimate ChIP enrichment for promoter proximal histone modifications. Use of different parameters for different antibodies (calling broad or narrow peaks) is possible. Additionally, users can elect to use BAM file from another experiment as control for MACS2 peak calling.
## Cite as
*Kartashov AV, Barski A. BioWardrobe: an integrated platform for analysis of epigenomics and transcriptomics data. Genome Biol. 2015;16(1):158. Published 2015 Aug 7. [doi:10.1186/s13059-015-0720-3](https://www.ncbi.nlm.nih.gov/pubmed/26248465)*
## Software versions
- Bowtie 1.2.0
- Samtools 1.4
- Preseq 2.0
- MACS2 2.1.1.20160309
- Bedtools 2.26.0
- UCSC userApps v358
## Inputs
| ID | Label | Description | Required | Default | Upstream analyses |
| ------------------------- | ---------------------------------------------- | ---------------------------------------------------------------------------------------------------------------------------------------------------------------- | :------: | ------- | ------------------------------- |
| **fastq\_file** | FASTQ file | Single-read sequencing data in FASTQ format (fastq, fq, bzip2, gzip, zip) | + | | |
| **indices\_folder** | Genome indices | Directory with the genome indices generated by Bowtie | + | | genome\_indices/bowtie\_indices |
| **annotation\_file** | Genome annotation file | Genome annotation file in TSV format | + | | genome\_indices/annotation |
| **genome\_size** | Effective genome size | The length of the mappable genome (hs, mm, ce, dm or number, for example 2.7e9) | + | | genome\_indices/genome\_size |
| **chrom\_length** | Chromosome lengths file | Chromosome lengths file in TSV format | + | | genome\_indices/chrom\_length |
| **broad\_peak** | Call broad peaks | Make MACS2 call broad peaks by linking nearby highly enriched regions | + | | |
| **control\_file** | Control ChIP-Seq single-read experiment | Indexed BAM file from the ChIP-Seq single-read experiment to be used as a control for MACS2 peak calling | | Null | control\_file/bambai\_pair |
| **exp\_fragment\_size** | Expected fragment size | Expected fragment size for read extenstion towards 3' end if *force\_fragment\_size* was set to True or if calculated by MACS2 fragment size was less that 80 bp | | 150 | |
| **force\_fragment\_size** | Force peak calling with expected fragment size | Make MACS2 don't build the shifting model and use expected fragment size for read extenstion towards 3' end | | False | |
| **clip\_3p\_end** | Clip from 3' end | Number of base pairs to clip from 3' end | | 0 | |
| **clip\_5p\_end** | Clip from 5' end | Number of base pairs to clip from 5' end | | 0 | |
| **remove\_duplicates** | Remove PCR duplicates | Remove PCR duplicates from sorted BAM file | | False | |
| **threads** | Number of threads | Number of threads for those steps that support multithreading | | 2 | |
## Outputs
| ID | Label | Description | Required | Visualization |
| ------------------------ | ---------------------------------- | ------------------------------------------------------------------------------------ | :------: | ------------------------------------------------------------------ |
| **fastx\_statistics** | FASTQ quality statistics | FASTQ quality statistics in TSV format | + | *Base Frequency* and *Quality Control* plots in *QC Plots* tab |
| **bambai\_pair** | Aligned reads | Coordinate sorted BAM alignment and index BAI files | + | *Nucleotide Sequence Alignments* track in *IGV Genome Browser* tab |
| **bigwig** | Genome coverage | Genome coverage in bigWig format | + | *Genome Coverage* track in *IGV Genome Browser* tab |
| **iaintersect\_result** | Gene annotated peaks | MACS2 peak file annotated with nearby genes | + | *Peak Coordinates* table in *Peak Calling* tab |
| **atdp\_result** | Average Tag Density Plot | Average Tag Density Plot file in TSV format | + | *Average Tag Density Plot* in *QC Plots* tab |
| **macs2\_called\_peaks** | Called peaks | Called peaks file with 1-based coordinates in XLS format | + | |
| **macs2\_narrow\_peaks** | Narrow peaks | Called peaks file in ENCODE narrow peak format | | *Narrow peaks* track in *IGV Genome Browser* tab |
| **macs2\_broad\_peaks** | Broad peaks | Called peaks file in ENCODE broad peak format | | *Broad peaks* track in *IGV Genome Browser* tab |
| **preseq\_estimates** | Expected Distinct Reads Count Plot | Expected distinct reads count file from Preseq in TSV format | | *Expected Distinct Reads Count Plot* in *QC Plots* tab |
| **workflow\_statistics** | Workflow execution statistics | Overall workflow execution statistics from bowtie\_aligner and samtools\_rmdup steps | + | *Overview* tab and experiment's preview |
| **bowtie\_log** | Read alignment log | Read alignment log file from Bowtie | + | |