import org.broadinstitute.sting.queue.extensions.gatk._ import org.broadinstitute.sting.queue.extensions.picard.PicardBamFunction import org.broadinstitute.sting.queue.QScript import org.broadinstitute.sting.queue.function.ListWriterFunction import scala.io.Source class dataProcessing extends QScript { qscript => @Input(doc="path to GenomeAnalysisTK.jar", shortName="gatk", required=true) var GATKjar: File = _ @Input(doc="path to AnalyzeCovariates.jar", shortName="ac", required=true) var ACJar: File = _ @Input(doc="path to Picard's MarkDuplicates.jar", shortName="dedup", required=true) var dedupJar: File = _ @Input(doc="path to R resources folder inside the Sting repository", shortName="r", required=true) var R: String = _ @Input(doc="input BAM file - or list of BAM files", shortName="i", required=true) var input: String = _ @Input(doc="Reference fasta file", shortName="R", required=false) var reference: File = new File("/seq/references/Homo_sapiens_assembly19/v1/Homo_sapiens_assembly19.fasta") @Input(doc="dbsnp ROD to use (VCF)", shortName="D", required=false) var dbSNP: File = new File("/humgen/gsa-hpprojects/GATK/data/dbsnp_132_b37.leftAligned.vcf") @Input(doc="extra VCF files to use as reference indels for Indel Realignment", shortName="indels", required=false) var indels: File = new File("/humgen/gsa-hpprojects/GATK/data/Comparisons/Unvalidated/AFR+EUR+ASN+1KG.dindel_august_release_merged_pilot1.20110126.sites.vcf") @Input(doc="the project name determines the final output (BAM file) base name. Example NA12878 yields NA12878.processed.bam", shortName="p", required=false) var projectName: String = "combined" @Input(doc="output path", shortName="outputDir", required=false) var outputDir: String = "" @Input(doc="the -L interval string to be used by GATK - output bams at interval only", shortName="L", required=false) var intervalString: String = "" @Input(doc="output bams at intervals only", shortName="intervals", required=false) var intervals: File = _ val queueLogDir: String = ".qlog/" // Simple boolean definitions for code clarity val knownsOnly: Boolean = true val intermediate: Boolean = true // General arguments to all programs trait CommandLineGATKArgs extends CommandLineGATK { this.jarFile = qscript.GATKjar this.reference_sequence = qscript.reference this.memoryLimit = 4 this.isIntermediate = true } def script = { var perLaneBamList: List[String] = Nil var recalibratedBamList: List[File] = Nil // Helpful variables val outName: String = qscript.projectName val outDir: String = qscript.outputDir // BAM files generated by the pipeline val bamList: String = outDir + outName + ".list" val joinedBams: String = outDir + outName + ".join.bam" val cleanedBam: String = outDir + outName + ".clean.bam" val fixedBam: String = outDir + outName + ".processed.bam" // Accessory files val knownTargetIntervals: String = outDir + outName + ".known_indels.intervals" val allTargetIntervals: String = outDir + outName + ".all_indels.intervals" add(new knownTargets(knownTargetIntervals)) // Populates the list of per lane bam files to process (single bam or list of bams). if (input.endsWith("bam")) perLaneBamList :+= input else for (bam <- Source.fromFile(input).getLines()) perLaneBamList :+= bam perLaneBamList.foreach { perLaneBam => // Helpful variables val baseName: String = swapExt(new File(perLaneBam.substring(perLaneBam.lastIndexOf("/")+1)), ".bam", "").toString() val baseDir: String = perLaneBam.substring(0, perLaneBam.lastIndexOf("/")+1) // BAM files generated by the pipeline val cleanedBam: String = baseName + ".clean.bam" val dedupedBam: String = baseName + ".clean.dedup.bam" val recalBam: String = baseName + ".clean.dedup.recal.bam" // Accessory files val metricsFile: String = baseName + ".metrics" val preRecalFile: String = baseName + ".pre_recal.csv" val postRecalFile: String = baseName + ".post_recal.csv" val preOutPath: String = baseName + ".pre" val postOutPath: String = baseName + ".post" add(new clean(perLaneBam, knownTargetIntervals, cleanedBam, knownsOnly, intermediate), new dedup(cleanedBam, dedupedBam, metricsFile), new cov(dedupedBam, preRecalFile), new recal(dedupedBam, preRecalFile, recalBam), new cov(recalBam, postRecalFile), new analyzeCovariates(preRecalFile, preOutPath), new analyzeCovariates(postRecalFile, postOutPath)) recalibratedBamList :+= new File(recalBam) } add(new writeList(recalibratedBamList, bamList), new joinBams(bamList, joinedBams), new allTargets(joinedBams, allTargetIntervals), new clean(joinedBams, allTargetIntervals, cleanedBam, !knownsOnly, !intermediate)) } class TargetBase (outIntervals: String) extends RealignerTargetCreator with CommandLineGATKArgs { this.out = new File(outIntervals) this.mismatchFraction = 0.0 this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.rodBind :+= RodBind("indels", "VCF", indels) } class knownTargets (outIntervals: String) extends TargetBase(outIntervals) { this.jobName = queueLogDir + outIntervals + ".ktarget" } class allTargets (inBams: String, outIntervals: String) extends TargetBase(outIntervals) { this.input_file :+= new File(inBams) this.memoryLimit = 6 this.jobName = queueLogDir + outIntervals + ".atarget" } class clean (inBams: String, tIntervals: String, outBam: String, knownsOnly: Boolean, intermediate: Boolean) extends IndelRealigner with CommandLineGATKArgs { this.input_file :+= new File(inBams) this.targetIntervals = new File(tIntervals) this.out = new File(outBam) this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.rodBind :+= RodBind("indels", "VCF", qscript.indels) this.useOnlyKnownIndels = knownsOnly this.doNotUseSW = true this.baq = org.broadinstitute.sting.utils.baq.BAQ.CalculationMode.CALCULATE_AS_NECESSARY this.compress = 0 this.U = org.broadinstitute.sting.gatk.arguments.ValidationExclusion.TYPE.NO_READ_ORDER_VERIFICATION // todo -- update this with the last consensus between Tim, Matt and Eric. This is ugly! this.isIntermediate = intermediate this.jobName = queueLogDir + outBam + ".clean" if (!intermediate && !qscript.intervalString.isEmpty()) this.intervalsString ++= List(qscript.intervalString) if (!intermediate && qscript.intervals != null) this.intervals :+= qscript.intervals } class dedup (inBam: String, outBam: String, metricsFile: String) extends PicardBamFunction { @Input(doc="fixed bam") var clean: File = new File(inBam) @Output(doc="deduped bam") var deduped: File = new File(outBam) @Output(doc="deduped bam index") var dedupedIndex: File = new File(outBam + ".bai") @Output(doc="metrics file") var metrics: File = new File(metricsFile) override def inputBams = List(clean) override def outputBam = deduped override def commandLine = super.commandLine + " M=" + metricsFile + " CREATE_INDEX=true" sortOrder = null this.memoryLimit = 6 this.jarFile = qscript.dedupJar this.isIntermediate = true this.jobName = queueLogDir + outBam + ".dedup" } class cov (inBam: String, outRecalFile: String) extends CountCovariates with CommandLineGATKArgs { this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.covariate ++= List("ReadGroupCovariate", "QualityScoreCovariate", "CycleCovariate", "DinucCovariate") this.input_file :+= new File(inBam) this.recal_file = new File(outRecalFile) this.jobName = queueLogDir + outRecalFile + ".covariates" } class recal (inBam: String, inRecalFile: String, outBam: String) extends TableRecalibration with CommandLineGATKArgs { @Output(doc="recalibrated bam index") var recalIndex: File = new File(outBam + ".bai") this.input_file :+= new File (inBam) this.recal_file = new File(inRecalFile) this.out = new File(outBam) this.U = org.broadinstitute.sting.gatk.arguments.ValidationExclusion.TYPE.NO_READ_ORDER_VERIFICATION // todo -- update this with the last consensus between Tim, Matt and Eric. This is ugly! this.index_output_bam_on_the_fly = true this.jobName = queueLogDir + outBam + ".recalibration" } class analyzeCovariates (inRecalFile: String, outPath: String) extends AnalyzeCovariates { this.jarFile = qscript.ACJar this.resources = qscript.R this.recal_file = new File(inRecalFile) this.output_dir = outPath this.jobName = queueLogDir + inRecalFile + ".analyze_covariates" } class writeList(inBams: List[File], outBamList: String) extends ListWriterFunction { this.inputFiles = inBams this.listFile = new File(outBamList) this.jobName = queueLogDir + outBamList + ".bamList" } class joinBams(inBams: String, outBam: String) extends PrintReads { this.input_file :+= new File(inBams) this.out = new File(outBam) this.jobName = queueLogDir + inBams + ".joinBams" } }