#tearsheet (for cori's use) #New tearsheet generator .libPaths('/humgen/gsa-firehose2/pipeline/repositories/StingProduction/R/') suppressMessages(library(gplots)); suppressMessages(library(ReadImages)); suppressMessages(library(gsalib)); cmdargs = gsa.getargs( list( title = list(value=NA, doc="Title for the tearsheet"), tsv = list(value=NA, doc="pipeline tsv file"), evalroot = list(value=NA, doc="VariantEval file base (everything before the .eval)"), tearout = list(value=NA, doc="Output path for tearsheet PDF")#, #plotout = list(value=NA, doc="Output path for plot PDF") ), doc="Creates a tearsheet" ); read.delim(cmdargs$tsv)->settable squids<-unique(settable[,1]) lane<-data.frame() samp<-data.frame() for(squid in squids){ gsa.read.squidmetrics(squid, TRUE)->lanemetrics addlanes<-lanemetrics[which(lanemetrics$"Individual ID" %in% settable[,2]),] gsa.read.squidmetrics(squid, FALSE)->samplemetrics addsamps<-samplemetrics[which(samplemetrics$"Sample" %in% settable[,2]),] lane<-rbind(lane, addlanes) samp<-rbind(samp, addsamps) } gsa.read.gatkreport(paste(evalroot, ".eval", sep=""))->FCeval gsa.read.gatkreport(paste(evalroot, "extraFC.eval", sep=""))->FCeval gsa.read.gatkreport(paste(evalroot, "extraFI.eval", sep=""))->FIeval gsa.read.gatkreport(paste(evalroot, "extraSA.eval", sep=""))->SAeval tearsheet<-function(){ pdf(file= cmdargs$tearout, width=22, height=17, pagecentre=TRUE, pointsize=24) #define layout postable<-matrix(c(1, 1, 1, 1, rep(c(2, 2, 4, 4), 5), rep(c(3, 3, 4, 4), 3), rep(c(3,3,5,5), 5), 6,7,8,9), nrow=15, ncol=4, byrow=TRUE) layout(postable, heights=c(1, rep(.18, 13), 2), respect=FALSE) #prep for title bar drop<-read.jpeg(system.file("data", "tearsheetdrop.jpg", package="gsalib")) #plot title bar par(mar=c(0,0,0,0)) plot(drop) text(155, 50, cmdargs$title, family="serif", adj=c(0,0), cex=3, col=gray(.25)) # Project summary projects = paste(squids, collapse=", "); used_samples = nrow(settable); unused_samples = 0; sequencing_protocol = samp$Initiative[1] bait_design = samp$"Bait Set"[1] callable_target = samp$"Target Territory"[1] table1<-rbind(paste(used_samples," used samples/", unused_samples + used_samples," total samples", sep=""), sequencing_protocol, bait_design, callable_target) rownames(table1)<-c("Samples","Sequencing Initiative", "Bait Design","Callable Target") par(mar=c(0,0,1,0)) textplot(table1, col.rownames="darkblue", show.colnames=FALSE, cex=1.25, valign="top") title(main=sprintf("Project Summary (%s)\n", projects), family="sans", cex.main=1.25, line=-1) # Bases summary reads_per_lane_mean = format(mean(lane$"PF Reads (HS)", na.rm=TRUE), 8, 3,1, scientific=TRUE); reads_per_lane_sd = format(sd(lane$"PF Reads (HS)", na.rm=TRUE), 8, 3,1, scientific=TRUE); lanessum<-sprintf("%s +/- %s\n", reads_per_lane_mean, reads_per_lane_sd) used_bases_per_lane_mean = format(mean(lane$"PF HQ Aligned Q20 Bases", na.rm=TRUE),8, 3,1, scientific=TRUE); used_bases_per_lane_sd = format(sd(lane$"PF HQ Aligned Q20 Bases", na.rm=TRUE), 8, 3,1, scientific=TRUE); lanessum<-c(lanessum, sprintf("%s +/- %s\n", used_bases_per_lane_mean, used_bases_per_lane_sd)); target_coverage_mean = mean(na.omit(lane$"Mean Target Coverage")); target_coverage_sd = sd(na.omit(lane$"Mean Target Coverage")); lanessum<-c(lanessum, sprintf("%0.2fx +/- %0.2fx\n", target_coverage_mean, target_coverage_sd)); pct_loci_gt_10x_mean = mean(na.omit(lane$"Target Bases 10x %")); pct_loci_gt_10x_sd = sd(na.omit(lane$"Target Bases 10x %")); lanessum<-c(lanessum, sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_10x_mean, pct_loci_gt_10x_sd)); pct_loci_gt_20x_mean = mean(na.omit(lane$"Target Bases 20x %")); pct_loci_gt_20x_sd = sd(na.omit(lane$"Target Bases 20x %")); lanessum<-c(lanessum,sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_20x_mean, pct_loci_gt_20x_sd)); pct_loci_gt_30x_mean = mean(na.omit(lane$"Target Bases 30x %")); pct_loci_gt_30x_sd = sd(na.omit(lane$"Target Bases 30x %")); lanessum<-c(lanessum,sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_30x_mean, pct_loci_gt_30x_sd)); reads_per_sample_mean = format(mean(samp$"PF Reads", na.rm=TRUE), 8, 3,1, scientific=TRUE); reads_per_sample_sd = format(sd(samp$"PF Reads",na.rm=TRUE), 8, 3,1, scientific=TRUE); sampssum<-sprintf("%s +/- %s\n", reads_per_sample_mean, reads_per_sample_sd); used_bases_per_sample_mean = format(mean(samp$"PF HQ Aligned Q20 Bases", na.rm=TRUE),8, 3,1, scientific=TRUE); used_bases_per_sample_sd = format(sd(samp$"PF HQ Aligned Q20 Bases", na.rm=TRUE), 8, 3,1, scientific=TRUE); sampssum<-c(sampssum, sprintf("%s +/- %s\n", used_bases_per_sample_mean, used_bases_per_sample_sd)); target_coverage_mean = mean(na.omit(samp$"Mean Target Coverage")); target_coverage_sd = sd(na.omit(samp$"Mean Target Coverage")); sampssum<-c(sampssum, sprintf("%0.2fx +/- %0.2fx\n", target_coverage_mean, target_coverage_sd)); pct_loci_gt_10x_mean = mean(na.omit(samp$"Target Bases 10x %")); pct_loci_gt_10x_sd = sd(na.omit(samp$"Target Bases 10x %")); sampssum<-c(sampssum, sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_10x_mean, pct_loci_gt_10x_sd)); pct_loci_gt_20x_mean = mean(na.omit(samp$"Target Bases 20x %")); pct_loci_gt_20x_sd = sd(na.omit(samp$"Target Bases 20x %")); sampssum<-c(sampssum, sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_20x_mean, pct_loci_gt_20x_sd)); pct_loci_gt_30x_mean = mean(na.omit(samp$"Target Bases 30x %")); pct_loci_gt_30x_sd = sd(na.omit(samp$"Target Bases 30x %")); sampssum<-c(sampssum, sprintf("%0.2f%% +/- %0.2f%%\n", pct_loci_gt_30x_mean, pct_loci_gt_30x_sd)); table2<-cbind(lanessum, sampssum) used_lanes = length(unique(paste(lane$Flowcell, lane$Lane))); if(nrow(lane)>used_lanes){ colnames(table2)<-c("Per barcoded readgroup", "Per sample") } else{ colnames(table2)<-c("Per lane", "Per sample") } rownames(table2)<-c("Reads", "Used bases", "Average target coverage", "% loci covered to 10x", "% loci covered to 20x","% loci covered to 30x") par(mar=c(0,0,1,0)) textplot(table2, rmar=1, col.rownames="dark blue", cex=1.25, valign="top") title(main="Bases Summary", family="sans", cex.main=1.25, line=0) # Sequencing summary instrument <- c(); if(length(grep("AAXX", lane$Flowcell))>0){ instrument <- c(instrument, "Illumina GA2") } if(length(grep("ABXX", lane$Flowcell))>0){ instrument <- c(instrument, "Illumina HiSeq") } if(length(instrument)>1){ instrument<-paste(instrument[1], instrument[2], sep=" and ") } used_lanes = length(unique(paste(lane$Flowcell, lane$Lane))); unused_lanes_by_sequencing = 0; #can we get this? unused_lanes_by_analysis = 0; lanes_per_sample_mean = mean(table(lane$"External ID"), na.rm=TRUE); lanes_per_sample_sd = sd(table(lane$"External ID"), na.rm=TRUE); lanes_per_sample_median = median(table(lane$"External ID")); lanes_paired = length(unique(paste(subset(lane, lane$"Lane Type" == "Paired")$Flowcell, subset(lane, lane$"Lane Type" == "Paired")$Lane))); lanes_widowed = length(unique(paste(subset(lane, lane$"Lane Type" == "Widowed")$Flowcell, subset(lane, lane$"Lane Type" == "Widowed")$Lane))); lanes_single = length(unique(paste(subset(lane, lane$"Lane Type" == "Single")$Flowcell, subset(lane, lane$"Lane Type" == "Single")$Lane))); read_length_mean = mean(lane$"Mean Read Length (P)"); read_length_sd = sd(lane$"Mean Read Length (P)"); read_length_median = median(lane$"Mean Read Length (P)"); date = lane$"Run Date"; date = sort(as.Date(date, format="%d-%b-%y")); start_date = format(date[1], "%B %d, %Y"); end_date = format(date[length(date)], "%B %d, %Y"); if(nrow(lane)>used_lanes){ used_lanes<-paste(used_lanes, " (multiplexed; ", nrow(lane), " total barcoded readgroups)", sep="") } table3<-rbind(paste(instrument), used_lanes, sprintf("%s rejected by sequencing, %s by analysis\n", unused_lanes_by_sequencing, unused_lanes_by_analysis), sprintf("%0.1f +/- %0.1f lanes (median=%0.1f)\n", lanes_per_sample_mean, lanes_per_sample_sd, lanes_per_sample_median), sprintf("%s paired, %s widowed, %s single\n", lanes_paired, lanes_widowed, lanes_single), sprintf("%0.1f +/- %0.1f bases (median=%0.1f)\n", read_length_mean, read_length_sd, read_length_median), sprintf("\tSequencing dates: %s to %s\n", start_date, end_date)) rownames(table3)<-c("Sequencer", "Used lanes", "Unused lanes","Used lanes/sample", "Lane parities", "Read lengths", "Sequencing dates") par(mar=c(0,0,1,0)) textplot(table3, rmar=1, col.rownames="dark blue", show.colnames=FALSE, cex=1.25, valign="top") title(main="Sequencing Summary", family="sans", cex.main=1.25, line=0) # Variant summary eval.counts = basiceval$CountVariants eval.counts.all = subset(eval.counts, Novelty == "all")$nVariantLoci; eval.counts.known = subset(eval.counts,Novelty == "known")$nVariantLoci; eval.counts.novel = subset(eval.counts, Novelty == "novel")$nVariantLoci; eval.titv = basiceval$TiTvVariantEvaluator eval.titv.all = subset(eval.titv, Novelty == "all")$tiTvRatio; eval.titv.known = subset(eval.titv, Novelty == "known")$tiTvRatio; eval.titv.novel = subset(eval.titv, Novelty == "novel")$tiTvRatio; table4 = matrix(c(eval.counts.all, eval.counts.known, eval.counts.novel, eval.titv.all, eval.titv.known, eval.titv.novel, "3.0 - 3.2", "3.2 - 3.4", "2.7 - 3.0"), nrow=3); rownames(table4) = c("All", "Known", "Novel"); colnames(table4) = c("Found", "Ti/Tv ratio", "Expected Ti/Tv ratio"); par(mar=c(0,0,0,0)) textplot(table4, rmar=1, col.rownames="dark blue", cex=1.25, valign="top") title(main="Variant Summary", family="sans", cex.main=1.25, line=-2) eval.bysample = SAeval$CountVariants eval.bysample.all = subset(eval.bysample, Novelty == "all" & Sample != "all"); eval.bysample.known = subset(eval.bysample, Novelty == "known"& Sample != "all"); eval.bysample.novel = subset(eval.bysample, Novelty == "novel"& Sample != "all"); eval.bysampleTITV = SAeval$TiTvVariantEvaluator eval.bysampleTITV.all = subset(eval.bysampleTITV, Novelty == "all" & Sample != "all"); eval.bysampleTITV.known = subset(eval.bysampleTITV, Novelty == "known"& Sample != "all"); eval.bysampleTITV.novel = subset(eval.bysampleTITV, Novelty == "novel"& Sample != "all"); eval.ac = basiceval$SimpleMetricsByAC.metrics eval.ac.all = subset(eval.ac, Novelty == "all"); eval.ac.known = subset(eval.ac, Novelty == "known"); eval.ac.novel = subset(eval.ac, Novelty == "novel"); eval.func = FCeval$CountVariants par(mar=c(5, 5, 4, 2) + 0.1) boxplot(eval.bysampleTITV.all$tiTvRatio, eval.bysampleTITV.known$tiTvRatio, eval.bysampleTITV.novel$tiTvRatio, main="Ti/Tv by Sample", col=c("dark gray", "blue", "red"), names=c("All", "Known", "Novel"), ylab="TI/TV per sample", main="",cex=1.3, cex.lab=1.3, cex.axis=1.3); par(mar=c(7, 5, 4, 2) + 0.1) ind = order(eval.bysample.all$nVariantLoci); plot(c(1:length(eval.bysample.all$nVariantLoci)), eval.bysample.all$nVariantLoci[ind], xlab="", col="black", xaxt="n", cex=1.1, cex.lab=1.1, cex.axis=1.1, main="Variants per Sample", ylab="Number of variants\n(axis in log space)", bty="n", log="y",ylim=c(100, max(eval.bysample.all$nVariantLoci))); points(c(1:length(eval.bysample.known$nVariantLoci)), eval.bysample.known$nVariantLoci[ind], col="blue", cex=1.3); points(c(1:length(eval.bysample.novel$nVariantLoci)), eval.bysample.novel$nVariantLoci[ind], col="red", cex=1.3); legend("bottomleft", max(eval.bysample.all$nVariantLoci)/2, c("All", "Known", "Novel"), col=c("black", "blue", "red"), pt.cex=1.3, pch=21); axis(1, at=c(1:length(eval.bysample.all$nVariantLoci)), lab=eval.bysample.all$Sample[ind], cex=.7, las=2) par(mar=c(6, 5, 4, 2) + 0.1) plot(sort(eval.ac.all$AC), eval.ac.all$n[order(eval.ac.all$AC)], ylim=c(1, max(eval.ac$n)), col="black", type="l", lwd=2, cex=1.1, cex.lab=1.1, cex.axis=1.1, xlab="Allele count\n(axis in log space)", ylab="Number of variants\n(axis in log space)", main="Variants by Allele Count", log="xy", bty="n"); points(sort(eval.ac.known$AC), eval.ac.known$n[order(eval.ac.known$AC)], col="blue", type="l", lwd=2); points(sort(eval.ac.novel$AC), eval.ac.novel$n[order(eval.ac.novel$AC)], col="red", type="l", lwd=2); legend("bottomleft", c("All", "Known", "Novel"), col=c("black", "blue", "red"), lwd=2); par(mar=c(5, 5, 4, 2) + 0.1) barplot(eval.func$nVariantLoci[4:nrow(eval.func)], col=c("dark gray", "blue", "red"), space=c(.2,0,0), log="y", main="Variants by Functional Class", xlab="Functional Class", ylab="# Variants\n(axis in log space)") axis(1, at=c(1.5,5,8.5), lab=c("Missense", "Nonsense", "Silent"), cex=.5, tick=FALSE) legend("top", c("All", "Known", "Novel"), fill=c("dark gray", "blue", "red"), cex=.7); dev.off() } tearsheet()