246 lines
12 KiB
R
Executable File
246 lines
12 KiB
R
Executable File
#Before executing this file, save squid files as csv, then as tab deliminated files with only the column values as the header, change the format of all cells to numbers. Assign the path to these files to "samples" and "lanes" respectively.
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#testcomment
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args<-commandArgs(TRUE)
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lanes<-args[1]
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samples<-args[2]
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sample_sets<-args[3]
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eval<-args[4]
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noveltitv<-args[5]
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knowntitv<-args[6]
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DOC<-args[7]
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if(is.na(sample_sets)){
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print("Please specify sample set for file naming and press enter.")
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scan("stdin", what="character",n=1)->sample_sets
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print("Thanks!")
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}
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if(is.na(lanes) == FALSE && is.na(samples)==FALSE){
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#this makes a table & graphs using Picard data
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read.delim(file=lanes, header= TRUE)->bylane;
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read.delim(file=samples, header= TRUE)->bysample;
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#Calc by lane metrics
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attach(bylane);
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callable.target<-HS_TARGET_TERRITORY[1];
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singlelanes<-length(which(Lane.Type=="Single"));
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pairedlanes<-length(which(Lane.Type=="Paired"));
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mean.read.lane<-signif(mean(AL_TOTAL_READS, na.rm=TRUE));
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sd.read.lane<-signif(sd(AL_TOTAL_READS, na.rm=TRUE));
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mean.ub.lane<-signif(mean(HS_ON_TARGET_BASES, na.rm=TRUE));
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sd.ub.lane<-signif(sd(HS_ON_TARGET_BASES, na.rm=TRUE));
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mean.cov.lane<-round(mean(HS_MEAN_TARGET_COVERAGE, na.rm=TRUE));
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sd.cov.lane<-round(sd(HS_MEAN_TARGET_COVERAGE, na.rm=TRUE));
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mean.10x.lane<-round(mean(HS_PCT_TARGET_BASES_10X, na.rm=TRUE));
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mean.20x.lane<-round(mean(HS_PCT_TARGET_BASES_20X, na.rm=TRUE));
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mean.30x.lane<-round(mean(HS_PCT_TARGET_BASES_30X, na.rm=TRUE));
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sd.10x.lane<-round(sd(HS_PCT_TARGET_BASES_10X, na.rm=TRUE));
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sd.20x.lane<-round(sd(HS_PCT_TARGET_BASES_20X, na.rm=TRUE));
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sd.30x.lane<-round(sd(HS_PCT_TARGET_BASES_30X, na.rm=TRUE));
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names<-paste(Project, " ", External.ID, "-", Lane, sep="")
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#makes a plot of the number of SNPS called per lane
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library(graphics)
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pdf(file=paste(sample_sets, "_SNPS.pdf", sep=""), width=0.2*length(SNP_TOTAL_SNPS), height=0.1*length(SNP_TOTAL_SNPS))
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layout(matrix(c(1,1 , 2), 1, 3, byrow=FALSE), respect=TRUE)
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plot(1:length(SNP_TOTAL_SNPS), main="SNPs Called in Each Lane", SNP_TOTAL_SNPS, xlab="", ylab="SNPs Called in Lane", xaxt="n", pch=16, col="blue")
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axis(side=1, at=(1:length(SNP_TOTAL_SNPS)), labels=names, cex.axis=0.75, las=2)
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boxplot(SNP_TOTAL_SNPS, main="SNPs Called in Lane", ylab="SNPs Called")
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if(length(boxplot.stats(SNP_TOTAL_SNPS)$out)==0){
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mtext("No outliers", side=1, line=4)
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}else{
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mtext(paste("Outlier SNP call counts in ", length(boxplot.stats(SNP_TOTAL_SNPS)$out), "lanes"), side=1, line=4)
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}
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dev.off()
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#makes SNP plot in log scale
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pdf(file=paste(sample_sets, "_SNPS_log.pdf", sep=""), width=0.2*length(SNP_TOTAL_SNPS), height=0.1*length(SNP_TOTAL_SNPS))
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layout(matrix(c(1,1 , 2), 1, 3, byrow=FALSE), respect=TRUE)
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plot(1:length(SNP_TOTAL_SNPS), log(SNP_TOTAL_SNPS), main="SNPs Called in Each Lane", xlab="", ylab="Log(SNPs Called in Lane)", xaxt="n", pch=16, col="blue")
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par(ylog=TRUE)
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axis(side=1, at=(1:length(SNP_TOTAL_SNPS)), labels=names, cex.axis=0.75, las=2)
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boxplot(SNP_TOTAL_SNPS, main="SNPs Called in Lane", ylab="SNPs Called")
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if(length(boxplot.stats(SNP_TOTAL_SNPS)$out)==0){
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mtext("No outliers", side=1, line=4)
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}else{
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mtext(paste("Outlier SNP call counts in ", length(boxplot.stats(SNP_TOTAL_SNPS)$out), "lanes"), side=1, line=4)
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}
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dev.off()
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#makes a plot of snp calls ordered by lane
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pdf(file=paste(sample_sets, "_SNPS_lane.pdf", sep=""), width=0.2*length(SNP_TOTAL_SNPS), height=0.1*length(SNP_TOTAL_SNPS))
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layout(matrix(c(1,1 , 2), 1, 3, byrow=FALSE), respect=TRUE)
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plot(1:length(SNP_TOTAL_SNPS), SNP_TOTAL_SNPS[order(Lane)], main="SNPs Called in Each Lane", xlab="", ylab="Log(SNPs Called in Lane)", xaxt="n", pch=16, col="blue")
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par(ylog=TRUE)
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axis(side=1, at=(1:length(SNP_TOTAL_SNPS)), labels=names[order(Lane)], cex.axis=0.75, las=2)
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boxplot(SNP_TOTAL_SNPS, main="SNPs Called in Lane", ylab="SNPs Called")
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if(length(boxplot.stats(SNP_TOTAL_SNPS)$out)==0){
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mtext("No outliers", side=1, line=4)
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}else{
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mtext(paste("Outlier SNP call counts in ", length(boxplot.stats(SNP_TOTAL_SNPS)$out), "lanes"), side=1, line=4)
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}
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dev.off()
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#makes a plot of fingerprint calls and labels them good or bad
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badsnps<-union(which(FP_CONFIDENT_MATCHING_SNPS<15), which(FP_CONFIDENT_MATCHING_SNPS<15))
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colors<-c(rep("Blue", length(FP_CONFIDENT_CALLS)))
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colors[badsnps]<-"Red"
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pdf(file=paste(sample_sets, "_Fingerprints.pdf", sep=""), width=.2*length(FP_CONFIDENT_CALLS), height=.1*length(FP_CONFIDENT_CALLS))
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par(mar=c(6, 4, 5, 4))
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plot(1:length(FP_CONFIDENT_MATCHING_SNPS), FP_CONFIDENT_MATCHING_SNPS, pch=16, ylim=c(0,24), ylab="Fingerprint calls", xlab="", xaxt="n", col=colors, main="Fingerprint Calling and Matching")
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points(1:length(FP_CONFIDENT_MATCHING_SNPS), FP_CONFIDENT_CALLS, col=colors)
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axis(side=1, at=(1:length(FP_CONFIDENT_CALLS)), labels=names, cex.axis=0.75, las=2)
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if(length(badsnps)>0){
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legend("bottomright", legend=c("Confident calls at fingerprint sites by lane", "Confident matching calls at fingerprint sites by lane", "Confident calls in bad lanes", "Confident matching calls in bad lanes"), pch=c(1, 16, 1, 16), col=c("Blue", "Blue", "Red", "Red"))
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mtext("Some problematic fingerprint sites", side=3)
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}else{
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legend("bottomright", legend=c("Confident calls at fingerprint sites by lane", "Confident matching calls at fingerprint sites by lane"), pch=c(1, 16), col="Blue")
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}
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dev.off()
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detach(bylane)
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#Calc by sample metrics
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attach(bysample);
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mean.lanes.samp<-signif(mean(X..Lanes.included.in.aggregation, na.rm = TRUE));
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sd.lanes.samp<-signif(sd(X..Lanes.included.in.aggregation, na.rm=TRUE));
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mean.mrl.samp<-signif(mean(Mean.Read.Length, na.rm=TRUE));
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sd.mrl.samp<-signif(sd(Mean.Read.Length, na.rm=TRUE));
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mean.read.samp<-signif(mean(Total.Reads, na.rm=TRUE));
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sd.read.samp<-signif(sd(Total.Reads, na.rm=TRUE));
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mean.ub.samp<-signif(mean(On.Target.Bases..HS., na.rm=TRUE));
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sd.ub.samp<-signif(sd(On.Target.Bases..HS., na.rm=TRUE));
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mean.cov.samp<-round(mean(Mean.Target.Coverage..HS., na.rm=TRUE));
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sd.cov.samp<-round(sd(Mean.Target.Coverage..HS., na.rm=TRUE));
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mean.10x.samp<-round(mean(PCT.Target.Bases.10x..HS., na.rm=TRUE));
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mean.20x.samp<-round(mean(PCT.Target.Bases.20x..HS., na.rm=TRUE));
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mean.30x.samp<-round(mean(PCT.Target.Bases.30x..HS., na.rm=TRUE));
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sd.10x.samp<-round(sd(PCT.Target.Bases.10x..HS., na.rm=TRUE));
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sd.20x.samp<-round(sd(PCT.Target.Bases.20x..HS., na.rm=TRUE));
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sd.30x.samp<-round(sd(PCT.Target.Bases.30x..HS., na.rm=TRUE));
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detach(bysample);
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#print all of this stuff out in R.
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print(paste("Callable Target: ", callable.target, " bases", sep=""), quote = FALSE);
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print(paste("Used Lanes per Sample: ", mean.lanes.samp, " +/- ", sd.lanes.samp, sep=""), quote=FALSE);
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print(paste("Parities: ", singlelanes, " single lanes, ", pairedlanes, " paired lanes", sep=""), quote=FALSE);
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print(paste("Read Legnths: ", mean.mrl.samp, " +/- ", sd.mrl.samp, sep=""), quote = FALSE);
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print(paste("Reads per lane: ", mean.read.lane, " +/- ", sd.read.lane, sep=""), quote = FALSE);
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print(paste("Reads per sample: ", mean.read.samp, " +/- ", sd.read.samp, sep=""), quote = FALSE);
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print(paste("Used bases per lane: ", mean.ub.lane, " +/- ", sd.ub.lane, sep=""), quote = FALSE);
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print(paste("Used bases per sample: ", mean.ub.samp, " +/- ", sd.ub.samp, sep=""), quote = FALSE)
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print(paste("Average target coverage per lane: ", mean.cov.lane, " +/- ", sd.cov.lane, sep=""), quote = FALSE);
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print(paste("Average target coverage per sample: ", mean.cov.samp, " +/- ", sd.cov.samp, sep=""), quote = FALSE);
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print(paste("% loci covered to 10x per lane: ", mean.10x.lane, "% +/- ", sd.10x.lane, "%", sep=""), quote = FALSE)
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print(paste("% loci covered to 10x per sample: ", mean.10x.samp, " +/- ", sd.10x.samp, "%", sep=""), quote = FALSE)
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print(paste("% loci covered to 20x per lane: ", mean.20x.lane, "% +/- ", sd.20x.lane, "%", sep=""), quote = FALSE)
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print(paste("% loci covered to 20x per sample: ", mean.20x.samp, "% +/- ", sd.20x.samp, "%", sep=""), quote = FALSE)
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print(paste("% loci covered to 30x per lane: ", mean.30x.lane, "% +/- ", sd.30x.lane, "%", sep=""), quote = FALSE)
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print(paste("% loci covered to 30x per sample: ", mean.30x.samp, "% +/- ", sd.30x.samp, "%", sep=""), quote = FALSE)
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}else{
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print("Lane and Sample metrics file paths not provided")
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}
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#Makes Error Rate percycle graph
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if(is.na(eval)==FALSE){
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read.delim(eval, header=TRUE)[2:ncol(read.delim(eval, header=TRUE))]->errpercycle
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pdf(paste(sample_sets, "_errorrate_per_cycle.pdf", sep=""), width=6, height=5)
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crazies<-which(errpercycle[75,]>0.3) #this can be changed to any kind of filter for particular lanes
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colors<-rainbow(ncol(errpercycle), s=0.5, v=0.5)
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colors[crazies]<-rainbow(length(crazies))
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weights<-rep(1, ncol(errpercycle))
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weights[crazies]<-2
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matplot(errpercycle, type="l", lty="solid", col=colors, lwd=weights, main="Error Rate per Cycle", ylab="Error Rate", xlab="Cycle", ylim=c(0, 0.7))
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if(length(crazies)>0){
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legend("topleft", title="Unusual Lanes", legend=colnames(errpercycle)[crazies], lty="solid", lwd=2, col=colors[crazies], xjust=0.5)
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}else{
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legend("topleft", legend="No unusual lanes.", bty="n")
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}
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dev.off()
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}else{
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print("Error Rate Per Cycle file paths not provided")
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}
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#Makes TI/TV known v novel graph
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if(is.na(noveltitv)==FALSE && is.na(knowntitv) == FALSE){
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pdf(paste(sample_set, "_TiTv.pdf", sep=""), width=6, height=5)
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read.table(file=noveltitv, header=FALSE)->novels
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read.table(file=knowntitv, header=FALSE)->knowns
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plot(novels[,2], col="red", ylim=c(0, 3.5), main="Ti/Tv for Novel and Known SNP calls", ylab="Ti/Tv", xlab="", xaxt="n")
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points(knowns[,2], col="blue")
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axis(side=1, at=(1:length(novels[,2])), labels=novels[,1], cex.axis=1, las=2)
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legend("bottomright", legend=c("Known Variants", "Novel Variants"), col=c("blue", "red"), pch=1, xjust=0.5)
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mtext("Lower Ti/Tv ratios indicated more false positive SNP calls.", side=1)
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dev.off()
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}else{
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print("Transition/transversion ratio file paths not provided")
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}
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#Make DOC graph
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if(is.na(DOC)==FALSE){
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pdf(paste(sample_set, "_DOC.pdf", sep=""), width=6, height=5)
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as.matrix(as.vector(read.delim(DOC, header=TRUE)[,2:502]))->DOCdata
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DOCdata<-matrix(DOCdata*100/sum(DOCdata[1,]), nrow=501, ncol=29, byrow=TRUE)
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colnames(DOCdata)<-read.delim(DOC, header=TRUE)[,1]
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oddies<-which(apply(DOCdata, 2, max)>10) #can be assigned any particular heuristic
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ncolors<-rainbow(ncol(DOCdata), s=0.5, v=0.5)
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ncolors[oddies]<-rainbow(length(oddies))
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nweights<-rep(1, ncol(DOCdata))
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nweights[oddies]<-2
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matplot(DOCdata, type="l", main="Depth of Coverage by Sample", ylab="Percent bases covered to a given depth", xlab="log(Depth)", log="x", col=ncolors, lty="solid", lwd=nweights)
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if(length(oddies)>0){
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legend("topright", title="Unusual Cases", legend=colnames(DOCdata)[oddies], lty="solid", lwd=2, col=ncolors[oddies], xjust=0.5)
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}else{
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legend("topright", legend="No unusual cases.", bty="n")
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}
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dev.off()
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}else{
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print("Depth of Coverage filepath not provided")
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}
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