diff --git a/astrocyte_pkg.yml b/astrocyte_pkg.yml
index 7db5da7e1e95561cd005a3b725ddc8633fb76e13..6e15b213afac9f222e9cde55e60a63f72cbd7951 100644
--- a/astrocyte_pkg.yml
+++ b/astrocyte_pkg.yml
@@ -41,16 +41,14 @@ documentation_files:
workflow_modules:
- 'python/3.6.1-2-anaconda'
- 'trimgalore/0.4.1'
- - 'cutadapt/1.9.1'
- - 'fastqc/0.11.2'
- 'bwa/intel/0.7.12'
- - 'samtools/1.4.1'
- 'sambamba/0.6.6'
- 'bedtools/2.26.0'
- 'deeptools/2.5.0.1'
- 'phantompeakqualtools/1.2'
- 'macs/2.1.0-20151222'
- 'UCSC_userApps/v317'
+ - 'R/3.4.1-gccmkl'
# A list of parameters used by the workflow, defining how to present them,
# options etc in the web interface. For each parameter:
@@ -113,13 +111,14 @@ workflow_parameters:
description: |
A design file listing sample id, fastq files, corresponding control id
and additional information about the sample.
+ regex: ".*tsv"
- id: genome
type: select
choices:
- [ 'GRCh38', 'Human GRCh38']
- [ 'GRCh37', 'Human GRCh37']
- - [ 'GRCh38', 'Mouse GRCh38']
+ - [ 'GRCm38', 'Mouse GRCm38']
required: true
description: |
Reference species and genome used for alignment and subsequent analysis.
diff --git a/workflow/conf/biohpc.config b/workflow/conf/biohpc.config
index 647927efa383ced7c7bcf0eed33482f6df562091..032ea21c155d843c63babbc977244440a8dc9f34 100644
--- a/workflow/conf/biohpc.config
+++ b/workflow/conf/biohpc.config
@@ -12,7 +12,7 @@ process {
cpus = 32
}
$alignReads{
- module = ['python/3.6.1-2-anaconda', 'bwa/intel/0.7.12', 'samtools/intel/1.3']
+ module = ['python/3.6.1-2-anaconda', 'bwa/intel/0.7.12', 'samtools/1.4.1']
cpus = 32
}
$filterReads{
@@ -47,6 +47,11 @@ process {
module = ['python/3.6.1-2-anaconda', 'bedtools/2.26.0']
executor = 'local'
}
+ $consensusPeaks {
+ module = ['R/3.4.1-gccmkl']
+ executor = 'local'
+ }
+
}
params {
diff --git a/workflow/main.nf b/workflow/main.nf
index 6da4f6dc8b3736d9cf5efd08a1987e9f9794e121..36a1bf92494d59615fae5bf07959e779c0115938 100644
--- a/workflow/main.nf
+++ b/workflow/main.nf
@@ -388,12 +388,14 @@ process peakAnnotation {
file consensusPeaks
- output:
- file "*chipseeker*" into chipseeker_originalpeak_output
- script:
- """
- module load python/2.7.x-anaconda
- module load R/3.3.2-gccmkl
- Rscript $baseDir/scripts/runChipseeker.R $design_file ${params.genomepath}
-"""
+ output:
+
+ file "*chipseeker*" into peakAnnotation
+
+ script:
+ """
+ module load python/2.7.x-anaconda
+ module load R/3.3.2-gccmkl
+ Rscript $baseDir/scripts/annotate_peaks.R $design_file $genome
+ """
}
diff --git a/workflow/scripts/annotate_peaks.R b/workflow/scripts/annotate_peaks.R
new file mode 100644
index 0000000000000000000000000000000000000000..74c60b3a137144c3335d1dc7a14ac45f3e308d7f
--- /dev/null
+++ b/workflow/scripts/annotate_peaks.R
@@ -0,0 +1,79 @@
+#!/bin/Rscript
+
+# Load libraries
+source("http://bioconductor.org/biocLite.R")
+if(!require("ChIPseeker")){
+ biocLite("ChIPseeker")
+ library("ChIPseeker")
+}
+
+# Currently mouse or human
+if (!require("TxDb.Hsapiens.UCSC.hg19.knownGene")){
+ biocLite("TxDb.Hsapiens.UCSC.hg19.knownGene")
+ library("TxDb.Hsapiens.UCSC.hg19.knownGene")
+}
+if (!require("TxDb.Mmusculus.UCSC.mm10.knownGene")){
+ biocLite("TxDb.Mmusculus.UCSC.mm10.knownGene")
+ library("TxDb.Mmusculus.UCSC.mm10.knownGene")
+}
+if (!require("TxDb.Hsapiens.UCSC.hg38.knownGene")){
+ biocLite("TxDb.Hsapiens.UCSC.hg38.knownGene")
+ library("TxDb.Hsapiens.UCSC.hg38.knownGene")
+}
+
+
+
+# Create parser object
+args <- commandArgs(trailingOnly=TRUE)
+
+# Check input args
+if (length(args) != 2) {
+ stop("Usage: annotate_peaks.r [ annotate_design.tsv ] [ genome_assembly ]", call.=FALSE)
+}
+
+design_file <- args[1]
+genome <-args[2]
+
+# Load UCSC Known Genes
+if(genome=='GRCh37') {
+ txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
+} else if(genome=='GRCm38') {
+ txdb <- TxDb.Mmusculus.UCSC.mm10.knownGene
+} else if(genome=='GRCh38') {
+ txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
+}
+
+# Load design file
+design <- read.csv(design_file, sep ='\t')
+files <- as.list(as.character(design$Peaks))
+names(files) <- design$Condition
+
+
+peakAnnoList <- lapply(files, annotatePeak, TxDb=txdb, tssRegion=c(-3000, 3000), verbose=FALSE)
+
+for(index in c(1:length(peakAnnoList))) {
+ filename <- paste(names(files)[index],".chipseeker_annotation.xls",sep="")
+ write.table(as.data.frame(peakAnnoList[[index]]),filename,sep="\t",quote=F)
+
+ # Draw individual plots
+
+ # Define names of Plots
+ pie_name <- paste(names(files)[index],".chipseeker_pie.pdf",sep="")
+ vennpie_name <- paste(names(files)[index],".chipseeker_vennpie.pdf",sep="")
+ upsetplot_name <- paste(names(files)[index],".chipseeker_upsetplot.pdf",sep="")
+
+ # Pie Plots
+ pdf(pie_name)
+ plotAnnoPie(peakAnnoList[[index]])
+ dev.off()
+
+ # Venn Diagrams
+ pdf(vennpie_name)
+ vennpie(peakAnnoList[[index]])
+ dev.off()
+
+ # Upset Plot
+ pdf(upsetplot_name)
+ upsetplot(peakAnnoList[[index]])
+ dev.off()
+}
diff --git a/workflow/scripts/overlap_peaks.py b/workflow/scripts/overlap_peaks.py
index 82f63b5ead547b93fa3b308692db2c1d2a7d3819..a5ca5c2ee49282abd330ab8b3394c2cce6df8586 100644
--- a/workflow/scripts/overlap_peaks.py
+++ b/workflow/scripts/overlap_peaks.py
@@ -166,7 +166,7 @@ def overlap(experiment, design):
os.remove(overlap_tr_fn)
os.remove(overlap_pr_fn)
- return overlapping_peaks_fn
+ return os.path.abspath(overlapping_peaks_fn)
def main():
@@ -186,14 +186,16 @@ def main():
design_diff = update_design(design_files_df)
# Make a design file for annotating Peaks
- design_anno = pd.DataFrame()
+ anno_cols = ['Condition', 'Peaks']
+ design_anno = pd.DataFrame(columns = anno_cols)
# Find consenus overlap peaks for each experiment
for experiment, df_experiment in design_peaks_df.groupby('experiment_id'):
replicated_peak = overlap(experiment, df_experiment)
design_diff.loc[design_diff.experiment_id == experiment, "peak"] = replicated_peak
+ design_anno.loc[experiment] = [experiment, replicated_peak]
- # Write out file
+ # Write out design files
design_diff.columns = ['SampleID',
'bamReads',
'Condition',
@@ -207,6 +209,7 @@ def main():
'PeakCaller']
design_diff.to_csv("design_diffPeaks.tsv", header=True, sep='\t', index=False)
+ design_anno.to_csv("design_annotatePeaks.tsv", header=True, sep='\t', index=False)
if __name__ == '__main__':
diff --git a/workflow/scripts/runChipseeker.R b/workflow/scripts/runChipseeker.R
deleted file mode 100644
index 8f8b0d011b511d8842c6d25d9842b3ef2a50974e..0000000000000000000000000000000000000000
--- a/workflow/scripts/runChipseeker.R
+++ /dev/null
@@ -1,51 +0,0 @@
-#!/bin/Rscript
-
-library(ChIPseeker)
-
-
-# Create parser object
-parser <- ArgumentParser()
-
-# Specify our desired options
-parser$add_argument("-d", "--design", help = "File path to design file", required = TRUE)
-parser$add_argument("-g", "--genome", help = "The genome assembly", required = TRUE)
-
-# Parse arguments
-args <- parser$parse_args()
-
-
-# Load UCSC Known Genes
-if(args$genome=='GRCh37') {
- library(TxDb.Hsapiens.UCSC.hg19.knownGene)
- txdb <- TxDb.Hsapiens.UCSC.hg19.knownGene
-} else if(args$genome=='GRCm38') {
- library(TxDb.Mmusculus.UCSC.mm10.knownGene)
- txdb <- TxDb.Mmusculus.UCSC.mm10.knownGene
-}
-else if(args$genome=='GRCh38') {
- library(TxDb.Hsapiens.UCSC.hg38.knownGene)
- txdb <- TxDb.Hsapiens.UCSC.hg38.knownGene
-}
-
-
-peakAnnoList <- lapply(files, annotatePeak, TxDb=txdb, tssRegion=c(-3000, 3000), verbose=FALSE)
-for(index in c(1:length(peakAnnoList)))
-{
- filename<-paste(names(files)[index],".chipseeker_annotation.xls",sep="")
- write.table(as.data.frame(peakAnnoList[[index]]),filename,sep="\t",quote=F)
- #draw individual plot
- pie_name <- paste(names(files)[index],".chipseeker_pie.pdf",sep="")
- vennpie_name <- paste(names(files)[index],".chipseeker_vennpie.pdf",sep="")
- upsetplot_name <- paste(names(files)[index],".chipseeker_upsetplot.pdf",sep="")
- pdf(pie_name)
- plotAnnoPie(peakAnnoList[[index]])
- dev.off()
- pdf(vennpie_name)
- vennpie(peakAnnoList[[index]])
- dev.off()
- pdf(upsetplot_name)
- upsetplot(peakAnnoList[[index]])
- dev.off()
-
-
-}