Snakemake workflow template for preparing single-end ChIP-Seq data
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This is a simple
snakemake
workflow template for preparing
single-end
ChIP-Seq data. The steps implemented are:
-
Download raw fastq files from SRA
-
Trim and Filter raw fastq files using
Code Snippets
16 17 18 19 20 21 22 23 24 25 26 | shell: """ AdapterRemoval \ --file1 {input.sample} \ {params.extra} \ {params.adapters} \ --threads {threads} \ --output1 {output.fastq} \ --discarded /dev/null \ --settings {output.settings} &> {log} """ |
12 13 14 15 16 17 18 19 20 21 22 | shell: """ ## Sort the file echo -e "Started sorting at $(date)" >> {log} sort \ -k1,1 -k2,2n \ -S {resources.mem_mb}M \ {input.bdg} | \ egrep $'^chr[0-9XY]+\t' > {output.bdg} echo -e "Finished sorting at $(date)" >> {log} """ |
38 39 40 41 42 43 | shell: """ echo -e "Started conversion at $(date)" >> {log} bedGraphToBigWig {input.bedgraph} {input.chrom_sizes} {output.bigwig} echo -e "Finished conversion at $(date)" >> {log} """ |
18 19 | script: "../scripts/bowtie2.py" |
35 36 37 38 39 40 41 42 43 44 45 46 | shell: """ DIR=$(dirname {output}) if [[ ! -d $DIR ]]; then mkdir -p $DIR fi bowtie2-inspect --summary {params.ref} | \ egrep '^Sequence' | \ sed -r 's/ /\\t/g' | \ cut -f2,4 > {output} """ |
15 16 17 18 19 20 21 22 | shell: """ fastqc \ {params.extra} \ -t {threads} \ --outdir {params.outdir} \ {input.fq} &> {log} """ |
72 73 74 75 76 77 78 79 80 81 82 | shell: """ macs2 callpeak \ -t {input.bam}\ -c {input.control} \ -f BAM --bdg --SPMR \ {params.extra} \ -n {params.prefix} \ --outdir {params.outdir} 2> {log} cp {log} {output.log} """ |
113 114 115 116 117 118 119 120 121 122 123 | shell: """ macs2 callpeak \ -t {input.bam}\ -c {input.control} \ -f BAM --bdg --SPMR \ {params.extra} \ -n {params.prefix} \ --outdir {params.outdir} 2> {log} cp {log} {output.log} """ |
149 150 151 152 153 154 155 156 | shell: """ macs2 bdgcmp \ -t {input.treatment} \ -c {input.control} \ {params} \ -o {output} 2> {log} """ |
169 170 171 172 | shell: """ Rscript --vanilla {input.script} {input.log} {output} >> {log} 2>&1 """ |
17 18 19 20 21 22 23 24 25 | shell: """ Rscript --vanilla \ {input.script} \ {input.bam} \ {input.chrom_sizes} \ {params.genome} \ {output.greylist} >> {log} 2>&1 """ |
17 18 19 20 21 22 23 24 25 | shell: """ multiqc \ {params.extra} \ --force \ -o {params.outdir} \ -n multiqc.html \ {input} 2> {log} """ |
16 17 18 19 20 21 22 | shell: """ Rscript --vanilla \ {input.script} \ {input.bam} \ {output} >> {log} 2>&1 """ |
40 41 42 43 44 45 46 47 | shell: """ Rscript --vanilla \ {input.script} \ {input.peaks} \ {input.bam} \ {output} >> {log} 2>&1 """ |
14 15 | script: "../scripts/picard_markduplicates.py" |
11 12 | script: "../scripts/create_site_yaml.R" |
33 34 35 36 | shell: """ R -e "rmarkdown::render_site('{input.rmd}')" >> {log} 2>&1 """ |
56 57 58 59 | shell: """ R -e "rmarkdown::render_site('{input.rmd}')" >> {log} 2>&1 """ |
83 84 85 86 | shell: """ R -e "rmarkdown::render_site('{input.rmd}')" >> {log} 2>&1 """ |
107 108 109 110 | shell: """ R -e "rmarkdown::render_site('{input.rmd}')" >> {log} 2>&1 """ |
123 124 125 126 127 128 129 130 131 132 133 | shell: """ ## Create the generic markdown Rscript --vanilla \ {input.r} \ {wildcards.target} \ {output.rmd} >> {log} 2>&1 ## Add the module directly as literal code cat {input.rmd} >> {output.rmd} """ |
161 162 163 164 | shell: """ R -e "rmarkdown::render_site('{input.rmd}')" >> {log} 2>&1 """ |
19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 | shell: """ TEMPDIR=$(mktemp -d -t samXXXXXXXXXX) echo -e "Writing to $TEMPDIR" >>{log} samtools view -h {params.view} {input} |\ samtools sort {params.sort} \ -@ {threads} -m 4G \ -T $TEMPDIR \ -O BAM \ -o {output} 2>> {log} echo -e "Deleting $TEMPDIR" >> {log} rm -rf $TEMPDIR """ |
46 47 48 49 | shell: """ samtools index -@ {threads} {input} {output} """ |
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 | __author__ = "Johannes Köster" __copyright__ = "Copyright 2016, Johannes Köster" __email__ = "koester@jimmy.harvard.edu" __license__ = "MIT" import os from snakemake.shell import shell from snakemake_wrapper_utils.samtools import get_samtools_opts def get_format(path: str) -> str: """ Return file format since Bowtie2 reads files that could be gzip'ed (extension: .gz) or bzip2'ed (extension: .bz2). """ if path.endswith((".gz", ".bz2")): return path.split(".")[-2].lower() return path.split(".")[-1].lower() bowtie2_threads = snakemake.threads - 1 if bowtie2_threads < 1: raise ValueError( f"This wrapper expected at least two threads, got {snakemake.threads}" ) # Setting parse_threads to false since samtools performs only # bam compression. Thus the wrapper would use *twice* the amount # of threads reserved by user otherwise. samtools_opts = get_samtools_opts(snakemake, parse_threads=False) extra = snakemake.params.get("extra", "") log = snakemake.log_fmt_shell(stdout=True, stderr=True) n = len(snakemake.input.sample) assert ( n == 1 ), "input->sample must have 1 (single-end) element." reads = "" if get_format(snakemake.input.sample[0]) in ("bam", "sam"): reads = f"-b {snakemake.input.sample[0]}" else: if snakemake.params.get("interleaved", False): reads = f"--interleaved {snakemake.input.sample[0]}" else: reads = f"-U {snakemake.input.sample[0]}" if all(get_format(sample) in ("fastq", "fq") for sample in snakemake.input.sample): extra += " -q " elif all(get_format(sample) == "tab5" for sample in snakemake.input.sample): extra += " --tab5 " elif all(get_format(sample) == "tab6" for sample in snakemake.input.sample): extra += " --tab6 " elif all( get_format(sample) in ("fa", "mfa", "fasta") for sample in snakemake.input.sample ): extra += " -f " metrics = snakemake.output.get("metrics") if metrics: extra += f" --met-file {metrics} " unaligned = snakemake.output.get("unaligned") if unaligned: extra += f" --un {unaligned} " unpaired = snakemake.output.get("unpaired") if unpaired: extra += f" --al {unpaired} " unconcordant = snakemake.output.get("unconcordant") if unconcordant: extra += f" --un-conc {unconcordant} " concordant = snakemake.output.get("concordant") if concordant: extra += f" --al-conc {concordant} " index = os.path.commonprefix(snakemake.input.idx).rstrip(".") shell( "(bowtie2" " --threads {bowtie2_threads}" " {reads} " " -x {index}" " {extra}" "| samtools view --with-header " " {samtools_opts}" " -" ") {log}" ) |
11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 | conda_pre <- system2("echo", "$CONDA_PREFIX", stdout = TRUE) if (conda_pre != ""){ conda_lib_path <- file.path(conda_pre, "lib", "R", "library") if (!dir.exists(conda_lib_path)) conda_lib_path <- NULL prev_paths <- .libPaths() paths_to_set <- unique(c(conda_lib_path, prev_paths)) .libPaths(paths_to_set) } library(tidyverse) library(yaml) library(glue) library(magrittr) config <- read_yaml(here::here("config", "config.yml")) samples <- read_tsv(here::here(config$samples)) targets <- sort(unique(samples$target)) dir <- basename(getwd()) site_yaml <- list( name = dir, output_dir = "../docs", navbar = list( title = dir, left = list( list(text = "Home", href = "index.html"), list( text = "QC", menu = list( list(text = "Raw Data", href = "raw_qc.html"), list(text = "Trimmed Data", href = "trimmed_qc.html"), list(text = "Alignments", href = "align_qc.html") ) ), list( text = "Results", menu = list() ) ), right = list( list(icon = "fa-github", href = "https://github.com/smped/prepareChIPs") ) ), output = list( html_document = list( code_folding = "hide", toc = TRUE, toc_float = TRUE, theme = "sandstone", highlight = "textmate", includes = list(after_body = "footer.html") ) ) ) ## Create the output directory, if it doesn't exist if (!is.null(site_yaml$output_dir)) { out_dir <- here::here("docs") message("Checking for directory: ", out_dir) if (!dir.exists(out_dir)) { message("Creating: ", out_dir) stopifnot(dir.create(out_dir)) } message(out_dir, " exists") } site_yaml$navbar$left[[3]]$menu <- lapply( targets, function(x) { list(text = x, href = glue("{x}_macs2_summary.html")) } ) ## Create the analysis directory if it doesn't exist analysis_dir <- here::here("analysis") if (!dir.exists(analysis_dir)) dir.create(analysis_dir) write_yaml(site_yaml, file.path(analysis_dir, "_site.yml")) |
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 | __author__ = "Johannes Köster, Derek Croote" __copyright__ = "Copyright 2020, Johannes Köster" __email__ = "johannes.koester@uni-due.de" __license__ = "MIT" import os import tempfile from snakemake.shell import shell from snakemake_wrapper_utils.snakemake import get_mem log = snakemake.log_fmt_shell(stdout=True, stderr=True) extra = snakemake.params.get("extra", "") # Parse memory mem_mb = get_mem(snakemake, "MiB") # Outdir outdir = os.path.dirname(snakemake.output[0]) if outdir: outdir = f"--outdir {outdir}" # Output compression compress = "" mem = f"-m{mem_mb}" if mem_mb else "" for output in snakemake.output: out_name, out_ext = os.path.splitext(output) if out_ext == ".gz": compress += f"pigz -p {snakemake.threads} {out_name}; " elif out_ext == ".bz2": compress += f"pbzip2 -p{snakemake.threads} {mem} {out_name}; " with tempfile.TemporaryDirectory() as tmpdir: mem = f"--mem {mem_mb}M" if mem_mb else "" shell( "(fasterq-dump --temp {tmpdir} --threads {snakemake.threads} {mem} " "{extra} {outdir} {snakemake.wildcards.accession}; " "{compress}" ") {log}" ) |
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 | __author__ = "Johannes Köster, Christopher Schröder" __copyright__ = "Copyright 2016, Johannes Köster" __email__ = "koester@jimmy.harvard.edu" __license__ = "MIT" import tempfile from pathlib import Path from snakemake.shell import shell from snakemake_wrapper_utils.java import get_java_opts from snakemake_wrapper_utils.samtools import get_samtools_opts, infer_out_format log = snakemake.log_fmt_shell() extra = snakemake.params.get("extra", "") # the --SORTING_COLLECTION_SIZE_RATIO default of 0.25 might # indicate a JVM memory overhead of that proportion java_opts = get_java_opts(snakemake, java_mem_overhead_factor=0.3) samtools_opts = get_samtools_opts(snakemake) tool = "MarkDuplicates" if snakemake.params.get("withmatecigar", False): tool = "MarkDuplicatesWithMateCigar" bams = snakemake.input.bams if isinstance(bams, str): bams = [bams] bams = list(map("--INPUT {}".format, bams)) output = snakemake.output.bam output_fmt = infer_out_format(output) convert = "" if output_fmt == "CRAM": output = "/dev/stdout" # NOTE: output format inference should be done by snakemake-wrapper-utils. Keeping it here for backwards compatibility. if snakemake.params.get("embed_ref", False): samtools_opts += ",embed_ref" convert = f" | samtools view {samtools_opts}" elif output_fmt == "BAM" and snakemake.output.get("idx"): extra += " --CREATE_INDEX" with tempfile.TemporaryDirectory() as tmpdir: shell( "(picard {tool}" # Tool and its subcommand " {java_opts}" # Automatic java option " {extra}" # User defined parmeters " {bams}" # Input bam(s) " --TMP_DIR {tmpdir}" " --OUTPUT {output}" # Output bam " --METRICS_FILE {snakemake.output.metrics}" # Output metrics " {convert}) {log}" # Logging ) output_prefix = Path(snakemake.output.bam).with_suffix("") if snakemake.output.get("idx"): if output_fmt == "BAM" and snakemake.output.idx != str(output_prefix) + ".bai": shell("mv {output_prefix}.bai {snakemake.output.idx}") |
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Created: 1yr ago
Updated: 1yr ago
Maitainers:
public
URL:
https://doi.org/10.48546/workflowhub.workflow.528.1
Name:
preparechips
Version:
v0.1.1
Downloaded:
0
Copyright:
Public Domain
License:
None
Keywords:
- Future updates
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