suppressMessages(library(gsalib)); suppressMessages(library(ROracle)); cmdargs = getargs( list( bamlist = list(value=NA, doc="list of BAM files"), evalroot = list(value=NA, doc="VariantEval root"), statsout = list(value=NA, doc="Output path for stats"), plotout = list(value=NA, doc="Output path for PDF") ), doc="Creates a tearsheet" ); #if (0) { bamlist = read.table(cmdargs$bamlist); fclanes = c(); for (bam in bamlist$V1) { bamheader = system(paste("samtools view -H", bam), intern=TRUE); if (length(bamheader) > 0) { rgs = bamheader[grep("^@RG", bamheader)]; for (rg in rgs) { id = grep("ID:", unlist(strsplit(rg, "\t")), value=TRUE); id = sub("ID:", "", id); fclanes = c(fclanes, id); } } else { cat(sprintf("Could not load '%s'\n", bam)); } } drv = dbDriver("Oracle"); con = dbConnect(drv, "REPORTING/REPORTING@ora01:1521/SEQPROD"); rs = dbSendQuery(con, statement = paste("SELECT * FROM ILLUMINA_PICARD_METRICS")); d = fetch(rs, n=-1); dbHasCompleted(rs); dbClearResult(rs); rs2 = dbSendQuery(con, statement = paste("SELECT * FROM ILLUMINA_SAMPLE_STATUS_AGG")); d2 = fetch(rs2, n=-1); dbHasCompleted(rs2); dbClearResult(rs2); oraCloseDriver(drv); squid_fclanes = sprintf("%s.%s", d$"Flowcell", d$"Lane"); squid_fclanes = gsub("A.XX", "", squid_fclanes); dproj = d[which(squid_fclanes %in% fclanes),]; d2proj = d2[which(d2$"Project" %in% unique(dproj$"Project") & d2$"Sample" %in% dproj$"External ID"),]; #} ## Tear sheet components # Project summary projects = paste(unique(dproj$"Project"), collapse=", "); cat(sprintf("Project Summary (%s)\n", projects), file=cmdargs$statsout, append=FALSE); used_samples = length(unique(dproj$"External ID")); cat(sprintf("\tUsed samples: %s\n", used_samples), file=cmdargs$statsout, append=TRUE); unused_samples = 0; cat(sprintf("\tUnused samples: %s\n", unused_samples), file=cmdargs$statsout, append=TRUE); sequencing_protocol = "Hybrid selection"; cat(sprintf("\tSequencing protocol: %s\n", sequencing_protocol), file=cmdargs$statsout, append=TRUE); bait_design = paste(unique(dproj$"Bait Set"), collapse=", "); cat(sprintf("\tBait design: %s\n", bait_design), file=cmdargs$statsout, append=TRUE); callable_target = paste(unique(dproj$"Target Territory"), collapse=", "); cat(sprintf("\tCallable target: %s\n", callable_target), file=cmdargs$statsout, append=TRUE); # Bases summary per lane cat("\nBases summary (excluding unused lanes/samples)\n", file=cmdargs$statsout, append=TRUE); cat("\tBases summary per lane\n", file=cmdargs$statsout, append=TRUE); reads_per_lane_mean = mean(dproj$"PF Reads (HS)"); reads_per_lane_sd = sd(dproj$"PF Reads (HS)"); cat(sprintf("\t\tReads: %s +/- %s\n", reads_per_lane_mean, reads_per_lane_sd), file=cmdargs$statsout, append=TRUE); used_bases_per_lane_mean = mean(dproj$"PF HQ Aligned Q20 Bases"); used_bases_per_lane_sd = sd(dproj$"PF HQ Aligned Q20 Bases"); cat(sprintf("\t\tUsed bases: %s +/- %s\n", used_bases_per_lane_mean, used_bases_per_lane_sd), file=cmdargs$statsout, append=TRUE); target_coverage_mean = mean(na.omit(dproj$"Mean Target Coverage")); target_coverage_sd = sd(na.omit(dproj$"Mean Target Coverage")); cat(sprintf("\t\tTarget coverage: %0.2fx +/- %0.2fx\n", target_coverage_mean, target_coverage_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_10x_mean = mean(na.omit(dproj$"Target Bases 10x %")); pct_loci_gt_10x_sd = sd(na.omit(dproj$"Target Bases 10x %")); cat(sprintf("\t\t%% loci > 10x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_10x_mean, pct_loci_gt_10x_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_20x_mean = mean(na.omit(dproj$"Target Bases 20x %")); pct_loci_gt_20x_sd = sd(na.omit(dproj$"Target Bases 20x %")); cat(sprintf("\t\t%% loci > 20x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_20x_mean, pct_loci_gt_20x_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_30x_mean = mean(na.omit(dproj$"Target Bases 30x %")); pct_loci_gt_30x_sd = sd(na.omit(dproj$"Target Bases 30x %")); cat(sprintf("\t\t%% loci > 30x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_30x_mean, pct_loci_gt_30x_sd), file=cmdargs$statsout, append=TRUE); cat("\tBases summary per sample\n", file=cmdargs$statsout, append=TRUE); reads_per_sample_mean = mean(d2proj$"PF Reads"); reads_per_sample_sd = sd(d2proj$"PF Reads"); cat(sprintf("\t\tReads: %s +/- %s\n", reads_per_sample_mean, reads_per_sample_sd), file=cmdargs$statsout, append=TRUE); used_bases_per_sample_mean = mean(d2proj$"PF HQ Aligned Q20 Bases"); used_bases_per_sample_sd = sd(d2proj$"PF HQ Aligned Q20 Bases"); cat(sprintf("\t\tUsed bases: %s +/- %s\n", used_bases_per_sample_mean, used_bases_per_sample_sd), file=cmdargs$statsout, append=TRUE); target_coverage_mean = mean(na.omit(d2proj$"Mean Target Coverage")); target_coverage_sd = sd(na.omit(d2proj$"Mean Target Coverage")); cat(sprintf("\t\tTarget coverage: %0.2fx +/- %0.2fx\n", target_coverage_mean, target_coverage_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_10x_mean = mean(na.omit(d2proj$"Target Bases 10x %")); pct_loci_gt_10x_sd = sd(na.omit(d2proj$"Target Bases 10x %")); cat(sprintf("\t\t%% loci > 10x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_10x_mean, pct_loci_gt_10x_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_20x_mean = mean(na.omit(d2proj$"Target Bases 20x %")); pct_loci_gt_20x_sd = sd(na.omit(d2proj$"Target Bases 20x %")); cat(sprintf("\t\t%% loci > 20x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_20x_mean, pct_loci_gt_20x_sd), file=cmdargs$statsout, append=TRUE); pct_loci_gt_30x_mean = mean(na.omit(d2proj$"Target Bases 30x %")); pct_loci_gt_30x_sd = sd(na.omit(d2proj$"Target Bases 30x %")); cat(sprintf("\t\t%% loci > 30x covered: %0.2f%% +/- %0.2f%%\n", pct_loci_gt_30x_mean, pct_loci_gt_30x_sd), file=cmdargs$statsout, append=TRUE); # Sequencing summary cat("\nSequencing summary\n", file=cmdargs$statsout, append=TRUE); instrument = "Illumina GA2"; cat(sprintf("\tSequencer: %s\n", instrument), file=cmdargs$statsout, append=TRUE); used_lanes = nrow(dproj); cat(sprintf("\tUsed lanes: %s\n", used_lanes), file=cmdargs$statsout, append=TRUE); unused_lanes_by_sequencing = 0; unused_lanes_by_analysis = 0; cat(sprintf("\tUnused lanes: %s rejected by sequencing, %s by analysis\n", unused_lanes_by_sequencing, unused_lanes_by_analysis), file=cmdargs$statsout, append=TRUE); lanes_per_sample_mean = mean(table(dproj$"External ID")); lanes_per_sample_sd = sd(table(dproj$"External ID")); lanes_per_sample_median = median(table(dproj$"External ID")); cat(sprintf("\tLanes/sample: %0.1f +/- %0.1f lanes (median=%0.1f)\n", lanes_per_sample_mean, lanes_per_sample_sd, lanes_per_sample_median), file=cmdargs$statsout, append=TRUE); lanes_paired = nrow(subset(dproj, dproj$"Lane Type" == "Paired")); lanes_widowed = nrow(subset(dproj, dproj$"Lane Type" == "Widowed")); lanes_single = nrow(subset(dproj, dproj$"Lane Type" == "Single")); cat(sprintf("\tLane parities: %s paired, %s widowed, %s single\n", lanes_paired, lanes_widowed, lanes_single), file=cmdargs$statsout, append=TRUE); read_length_mean = mean(dproj$"Mean Read Length (P)"); read_length_sd = sd(dproj$"Mean Read Length (P)"); read_length_median = median(dproj$"Mean Read Length (P)"); cat(sprintf("\tRead length: %0.1f +/- %0.1f bases (median=%0.1f)\n", read_length_mean, read_length_sd, read_length_median), file=cmdargs$statsout, append=TRUE); date = dproj$"Run Date"; date = sub("JAN", "01", date); date = sub("FEB", "02", date); date = sub("MAR", "03", date); date = sub("APR", "04", date); date = sub("MAY", "05", date); date = sub("JUN", "06", date); date = sub("JUL", "07", date); date = sub("AUG", "08", date); date = sub("SEP", "09", date); date = sub("OCT", "10", date); date = sub("NOV", "11", date); date = sub("DEC", "12", date); date = date[order(as.Date(date, format="%d-%m-%Y"))]; start_date = date[1]; end_date = date[length(date)]; cat(sprintf("\tSequencing dates: %s to %s\n", start_date, end_date), file=cmdargs$statsout, append=TRUE); # Variant summary cat("\nVariant summary\n", file=cmdargs$statsout, append=TRUE); eval.counts = read.csv(paste(cmdargs$evalroot, ".Count_Variants.csv", sep=""), header=TRUE, comment.char="#"); eval.counts.called = subset(eval.counts, evaluation_name == "eval" & comparison_name == "dbsnp" & jexl_expression == "none" & filter_name == "called"); eval.counts.called.all = subset(eval.counts.called, novelty_name == "all")$nVariantLoci; eval.counts.called.known = subset(eval.counts.called, novelty_name == "known")$nVariantLoci; eval.counts.called.novel = subset(eval.counts.called, novelty_name == "novel")$nVariantLoci; eval.titv = read.csv(paste(cmdargs$evalroot, ".Ti_slash_Tv_Variant_Evaluator.csv", sep=""), header=TRUE, comment.char="#"); eval.titv.called = subset(eval.titv, evaluation_name == "eval" & comparison_name == "dbsnp" & jexl_expression == "none" & filter_name == "called"); eval.titv.called.all = subset(eval.titv.called, novelty_name == "all")$ti.tv_ratio; eval.titv.called.known = subset(eval.titv.called, novelty_name == "known")$ti.tv_ratio; eval.titv.called.novel = subset(eval.titv.called, novelty_name == "novel")$ti.tv_ratio; vars = matrix(c(eval.counts.called.all, eval.counts.called.known, eval.counts.called.novel, eval.titv.called.all, eval.titv.called.known, eval.titv.called.novel, "3.0 - 3.2", "3.2 - 3.4", "2.7 - 3.0"), nrow=3); rownames(vars) = c("All", "Known", "Novel"); colnames(vars) = c("Found", "Ti/Tv ratio", "Expected Ti/Tv ratio"); cat(sprintf("\t\tFound\tTi/Tv ratio\tExpected Ti/Tv ratio\n"), file=cmdargs$statsout, append=TRUE); cat(sprintf("\tAll\t%s\t%s\t\t%s\n", eval.counts.called.all, eval.titv.called.all, "3.0 - 3.2"), file=cmdargs$statsout, append=TRUE); cat(sprintf("\tKnown\t%s\t%s\t\t%s\n", eval.counts.called.known, eval.titv.called.known, "3.2 - 3.4"), file=cmdargs$statsout, append=TRUE); cat(sprintf("\tNovel\t%s\t%s\t\t%s\n", eval.counts.called.novel, eval.titv.called.novel, "2.7 - 3.0"), file=cmdargs$statsout, append=TRUE); # Plots eval.bysample = read.csv(paste(cmdargs$evalroot, ".SimpleMetricsBySample.csv", sep=""), header=TRUE, comment.char="#"); eval.bysample.called = subset(eval.bysample, evaluation_name == "eval" & comparison_name == "dbsnp" & jexl_expression == "none" & filter_name == "called"); eval.bysample.called.all = subset(eval.bysample.called, novelty_name == "all"); eval.bysample.called.known = subset(eval.bysample.called, novelty_name == "known"); eval.bysample.called.novel = subset(eval.bysample.called, novelty_name == "novel"); eval.ac = read.csv(paste(cmdargs$evalroot, ".MetricsByAc.csv", sep=""), header=TRUE, comment.char="#"); eval.ac.called = subset(eval.ac, evaluation_name == "eval" & comparison_name == "dbsnp" & jexl_expression == "none" & filter_name == "called"); eval.ac.called.all = subset(eval.ac.called, novelty_name == "all"); eval.ac.called.known = subset(eval.ac.called, novelty_name == "known"); eval.ac.called.novel = subset(eval.ac.called, novelty_name == "novel"); eval.func = read.csv(paste(cmdargs$evalroot, ".Functional_Class_Counts_by_Sample.csv", sep=""), header=TRUE, comment.char="#"); eval.func.called = subset(eval.func, evaluation_name == "eval" & comparison_name == "dbsnp" & jexl_expression == "none" & filter_name == "called"); eval.func.called.all = subset(eval.func.called, novelty_name == "all"); eval.func.called.known = subset(eval.func.called, novelty_name == "known"); eval.func.called.novel = subset(eval.func.called, novelty_name == "novel"); pdf(cmdargs$plotout); boxplot(eval.bysample.called.all$CountVariants, eval.bysample.called.known$CountVariants, eval.bysample.called.novel$CountVariants, names=c("All", "Known", "Novel"), ylab="Variants per sample", main="", cex=1.3, cex.lab=1.3, cex.axis=1.3); ind = order(eval.bysample.called.all$CountVariants); plot(c(1:length(eval.bysample.called.all$CountVariants)), eval.bysample.called.all$CountVariants[ind], col="black", cex=1.3, cex.lab=1.3, cex.axis=1.3, xlab="Sample", ylab="Number of variants", bty="n", ylim=c(0, max(eval.bysample.called.all$CountVariants))); points(c(1:length(eval.bysample.called.known$CountVariants)), eval.bysample.called.known$CountVariants[ind], col="blue", cex=1.3); points(c(1:length(eval.bysample.called.novel$CountVariants)), eval.bysample.called.novel$CountVariants[ind], col="red", cex=1.3); legend(0, max(eval.bysample.called.all$CountVariants)/2, c("All", "Known", "Novel"), col=c("black", "blue", "red"), pt.cex=1.3, pch=21); plot(eval.ac.called.all$AC, eval.ac.called.all$n, col="black", type="l", lwd=2, cex=1.3, cex.lab=1.3, cex.axis=1.3, xlab="Allele count", ylab="Number of variants", main="", log="xy", bty="n"); points(eval.ac.called.known$AC, eval.ac.called.known$n, col="blue", type="l", lwd=2); points(eval.ac.called.novel$AC, eval.ac.called.novel$n, col="red", type="l", lwd=2); legend("topright", c("All", "Known", "Novel"), col=c("black", "blue", "red"), lwd=2); plot(eval.func.called.all$Synonymous[ind] / (eval.func.called.all$Missense + eval.func.called.all$Nonsense)[ind], ylim=c(0, 2), cex=1.3, cex.lab=1.3, cex.axis=1.3, bty="n", xlab="Sample", ylab="Ratio of synonymous to non-synonymous variants", col="black"); points(eval.func.called.known$Synonymous[ind] / (eval.func.called.known$Missense + eval.func.called.known$Nonsense)[ind], cex=1.3, col="blue"); points(eval.func.called.novel$Synonymous[ind] / (eval.func.called.novel$Missense + eval.func.called.novel$Nonsense)[ind], cex=1.3, col="red"); legend("topright", c("All", "Known", "Novel"), col=c("black", "blue", "red"), pt.cex=1.3, pch=21); dev.off();