import net.sf.picard.reference.FastaSequenceFile import org.broadinstitute.sting.datasources.pipeline.Pipeline import org.broadinstitute.sting.queue.extensions.gatk._ import org.broadinstitute.sting.queue.extensions.samtools._ import org.broadinstitute.sting.queue.{QException, QScript} import collection.JavaConversions._ import org.broadinstitute.sting.utils.yaml.YamlUtils import org.broadinstitute.sting.utils.report.VE2ReportFactory.VE2TemplateType class WGSpipeline extends QScript { qscript => @Input(doc="File with list of Bams", shortName = "bams", required = true) var bamList: File = _ @Input(doc="path to GATK jar", shortName="gatk", required=true) var gatkJar: File = _ @Input(doc="Project Name", shortName = "P", required = true) var project: String =_ @Input(doc="path to tmp space for storing intermediate bam files", shortName="outputTmpDir", required=true) var outputTmpDir: String = _ @Input(doc="reference file", shortName = "R", required = true) var reference: File = _ @Input(doc="dbSNP", shortName = "D", required = true) var dbSNP: File = _ @Input(doc="gsa-pipeline directory key", shortName = "dirKey", required = true) var dirkey: String =_ @Input(doc="version id in format ### (ie 001)", shortName = "v", required = true) var version: String =_ @Input(doc="Flag for running the entire genome. Otherwise a QC run of chr 20 is all that is run.", shortName = "freeze", required = false) var freeze: Boolean = false private val dindelCalls: String = "/humgen/gsa-hpprojects/GATK/data/Comparisons/Unvalidated/AFR+EUR+ASN+1KG.dindel_august_release_merged_pilot1.20110126.sites.vcf" val hapmap = "/humgen/gsa-hpprojects/GATK/data/Comparisons/Validated/HapMap/3.3/sites_r27_nr.b37_fwd.vcf" val g1k = "/humgen/gsa-hpprojects/GATK/data/Comparisons/Unvalidated/1kg_pilot1_projectCalls/ALL.low_coverage.2010_07.hg19.vcf" val omni = "/humgen/gsa-hpprojects/GATK/data/Comparisons/Validated/Omni2.5_chip/Omni25_sites_1525_samples.b37.vcf" private var pipeline: Pipeline = _ trait CommandLineGATKArgs extends CommandLineGATK { this.jarFile = qscript.gatkJar this.reference_sequence = qscript.reference this.memoryLimit = Some(3) } def script = { var chr: Int = 20 var runType: String = "QC" if (qscript.freeze) { var chr: Int =20 var runType = "freeze" } //TODO: Impliment whole genome once infrastructure for waiting for tmp space is available. // for now this will only run on chromosome 20 because otherwise we might run into space errors. else{ var chr: Int = 20 var runType = "QC" } val dirbase = qscript.dirkey + "/" + runType + "_v" +qscript.version val base = dirbase + "/Chunks/" + qscript.project + "." + runType val projectBase = dirbase + "/" + qscript.project + "." + runType val basesPerJob: Int = 3000000 val lastBase: Int = 63025520 var start: Int = 1 var stop: Int = start - 1 + basesPerJob if( stop > lastBase ) { stop = lastBase } var jobNumber: Int = 1 var mergeList: List[List[_]] = Nil while( jobNumber < (lastBase.toFloat / basesPerJob.toFloat) + 1.0) { var interval = "%d:%d-%d".format(chr, start, stop) val baseTmpName: String = qscript.outputTmpDir + "/" +base + "chunk_" +jobNumber val baseJobName: String = qscript.project + "_chunk_" + jobNumber // 1.) Create cleaning targets val target = new RealignerTargetCreator with CommandLineGATKArgs target.memoryLimit = Some(3) target.input_file :+= qscript.bamList target.intervalsString :+= interval target.out = new File(baseTmpName + ".target.intervals") target.mismatchFraction = Some(0.0) target.maxIntervalSize = Some(700) target.rodBind :+= RodBind("indels1", "VCF", qscript.dindelCalls) target.jobName = baseJobName + ".target" target.jobOutputFile = new File(baseTmpName + ".target.out") target.isIntermediate = true // 2.) Clean without SW val clean = new IndelRealigner with CommandLineGATKArgs val cleanedBam = new File(baseTmpName + ".cleaned.bam") clean.memoryLimit = Some(4) clean.input_file :+= qscript.bamList clean.intervalsString :+= interval clean.targetIntervals = target.out clean.jobOutputFile = new File (baseTmpName + ".clean.out" ) clean.out = cleanedBam clean.doNotUseSW = true clean.simplifyBAM = true clean.rodBind :+= RodBind("indels1", "VCF", qscript.dindelCalls) clean.jobName = baseJobName + ".clean" clean.isIntermediate = true clean.compress = Some(0) clean.rodBind :+= RodBind("dbsnp", "VCF", qscript.dbSNP ) val call = new UnifiedGenotyper with CommandLineGATKArgs call.out = new File("/humgen/gsa-pipeline/"+base+ "chunk_" +jobNumber + ".vcf") call.dcov = Some( 50 ) call.stand_call_conf = Some( 4.0 ) call.stand_emit_conf = Some( 4.0 ) call.baq = org.broadinstitute.sting.utils.baq.BAQ.CalculationMode.CALCULATE_AS_NECESSARY call.jobName = baseJobName +".call" call.exactCalculation = org.broadinstitute.sting.gatk.walkers.genotyper.ExactAFCalculationModel.ExactCalculation.LINEAR_EXPERIMENTAL call.jobOutputFile = new File ("/humgen/gsa-pipeline/"+base+ "chunk_" +jobNumber + ".call.out" ) call.input_file = List(clean.out) call.intervalsString :+= interval call.rodBind :+= RodBind("dbsnp", "VCF", qscript.dbSNP ) mergeList:+= List(baseJobName, call.out) add(target, clean, call) start += basesPerJob stop += basesPerJob if( stop > lastBase ) { stop = lastBase } jobNumber += 1 } val combineVCFs = new CombineVariants with CommandLineGATKArgs combineVCFs.priority = "" for (c <- mergeList ){ combineVCFs.rodBind :+= RodBind(c(0).toString, "VCF", c(1).toString) combineVCFs.priority += c(0).toString +"," } combineVCFs.variantMergeOptions = org.broadinstitute.sting.gatk.contexts.variantcontext.VariantContextUtils.VariantMergeType.UNION combineVCFs.jobName = qscript.project + ".combine" combineVCFs.out = new File("/humgen/gsa-pipeline/" + projectBase + ".merged.vcf") combineVCFs.jobOutputFile = new File ("/humgen/gsa-pipeline/" +projectBase + ".combine.out") combineVCFs.genotypemergeoption = org.broadinstitute.sting.gatk.contexts.variantcontext.VariantContextUtils.GenotypeMergeType.PRIORITIZE combineVCFs.intervalsString :+= "20:1-63025520" ///TODO impliment only using the chr 20 or nothing if freeze combineVCFs.assumeIdenticalSamples = true val sv = new SelectVariants with CommandLineGATKArgs sv.selectIndels = true sv.out = swapExt(combineVCFs.out, "merged.vcf", "indelsOnly.vcf") sv.rodBind :+= RodBind("variant", "VCF", combineVCFs.out) sv.jobName = qscript.project + ".SV" sv.jobOutputFile = swapExt(combineVCFs.jobOutputFile, "combine.out", "sv.out") val filter = new VariantFiltration with CommandLineGATKArgs filter.intervalsString :+= "20:1-63025520" ///TODO impliment only using the chr 20 or nothing if freeze filter.variantVCF = sv.out filter.out = swapExt(sv.out, ".vcf", ".filtered.vcf") filter.filterName ++= List("HARD_TO_VALIDATE") //ToDO check to make sure that this is what Guillermo recomends filter.filterExpression ++= List("\"MQ0 >= 4 && (MQ0 / (1.0 * DP)) > 0.1\"") filter.jobName = qscript.project + ".VF" filter.jobOutputFile = swapExt(sv.jobOutputFile, ".sv.out", ".filter.out") val recombine = new CombineVariants with CommandLineGATKArgs recombine.rodBind :+= RodBind("indels", "VCF", sv.out) recombine.rodBind :+= RodBind("all", "VCF", combineVCFs.out) recombine.priority = "indels,all" recombine.genotypemergeoption = org.broadinstitute.sting.gatk.contexts.variantcontext.VariantContextUtils.GenotypeMergeType.PRIORITIZE recombine.out = swapExt(combineVCFs.out, "vcf", "filtered.vcf") recombine.jobName = qscript.project + ".recombine" recombine.jobOutputFile = swapExt(combineVCFs.jobOutputFile, "combine.out", "recombine.out") val cr = new ContrastiveRecalibrator with CommandLineGATKArgs cr.rodBind :+= RodBind("hapmap","VCF", qscript.hapmap, "training=true, prior=3.0") cr.rodBind :+= RodBind("1kg", "VCF", qscript.omni, "training=true, prior=3.0") cr.rodBind :+= RodBind("input", "VCF", recombine.out) cr.rodBind :+= RodBind("dbsnp", "VCF", qscript.dbSNP, "known=true") cr.use_annotation ++= List("QD", "SB", "HaplotypeScore", "HRun") cr.jobName = qscript.project + ".cr" cr.jobOutputFile = swapExt(combineVCFs.jobOutputFile, ".combine.out", ".cr.out") cr.intervalsString :+= "20" ///TODO impliment only using the chr 20 or nothing if freeze cr.allPoly = true cr.tranches_file =swapExt(filter.out, "merged.filtered.vcf", "tranches") cr.recal_file = swapExt(filter.out, "merged.filtered.vcf", "") cr.tranche ++= List("0.1","0.5","0.75","1.0","1.25","1.35","1.5","1.7","1.8","1.9","2.0","2.1","2.2","2.3","2.5","3.0","5.0","8.0","10.0") val ar = new ApplyRecalibration with CommandLineGATKArgs ar.tranches_file = cr.tranches_file ar.recal_file = cr.recal_file ar.ignore_filter++= List("HARD_TO_VALIDATE") ar.out = swapExt(filter.out, "vcf", "recalibrated.vcf") ar.jobName = qscript.project + ".ar" ar.jobOutputFile = swapExt(combineVCFs.jobOutputFile, ".combine.out", ".ar.out") val stdEval = new VariantEval with CommandLineGATKArgs stdEval.jobName = qscript.project+".eval" stdEval.jobOutputFile = swapExt(combineVCFs.jobOutputFile, ".combine.out", ".eval.out") stdEval.noST = true stdEval.noEV = true stdEval.evalModule ++= List("SimpleMetricsByAC", "TiTvVariantEvaluator", "CountVariants") stdEval.stratificationModule ++= List("EvalRod", "CompRod", "Novelty") stdEval.rodBind :+= RodBind("dbsnp", "VCF", qscript.dbSNP) stdEval.rodBind :+= RodBind("eval", "VCF", ar.out) stdEval.out = swapExt(ar.out, ".vcf", ".eval") stdEval.tranchesFile = cr.tranches_file add(combineVCFs, filter, sv, recombine, cr, ar, stdEval) } }