package oneoffs.carneiro 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 net.sf.samtools.{SAMFileReader,SAMFileHeader,SAMReadGroupRecord} import collection.JavaConversions._ import org.broadinstitute.sting.commandline.ArgumentSource class dataProcessingV2 extends QScript { qscript => @Input(doc="path to GenomeAnalysisTK.jar", fullName="path_to_gatk_jar", shortName="gatk", required=true) var GATKjar: File = _ @Input(doc="path to AnalyzeCovariates.jar", fullName="path_to_ac_jar", shortName="ac", required=true) var ACJar: File = _ @Input(doc="path to Picard's MarkDuplicates.jar", fullName="path_to_dedup_jar", shortName="dedup", required=true) var dedupJar: File = _ @Input(doc="path to Picard's MergeSamFiles.jar", fullName="path_to_merge_jar", shortName="merge", required=true) var mergeBamJar: File = _ @Input(doc="path to R resources folder inside the Sting repository", fullName="path_to_r", shortName="r", required=true) var R: String = _ @Input(doc="The path to the binary of bwa (usually BAM files have already been mapped - but if you want to remap this is the option)", fullName="path_to_bwa", shortName="bwa", required=false) var bwaPath: File = _ @Input(doc="input BAM file - or list of BAM files", fullName="input", shortName="i", required=true) var input: File = _ @Input(doc="Reference fasta file", fullName="reference", 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)", fullName="dbsnp", 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", fullName="extra_indels", 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", fullName="project", shortName="p", required=false) var projectName: String = "project" @Input(doc="Perform cleaning on knowns only", fullName="knowns_only", shortName="knowns", required=false) var knownsOnly: Boolean = false @Input(doc="Perform cleaning using Smith Waterman", fullName="use_smith_waterman", shortName="sw", required=false) var useSW: Boolean = false @Input(doc="Output path for the processed BAM files.", fullName="output_directory", shortName="outputDir", required=false) var outputDir: String = "" @Input(doc="the -L interval string to be used by GATK - output bams at interval only", fullName="gatk_interval_string", shortName="L", required=false) var intervalString: String = "" @Input(doc="an intervals file to be used by GATK - output bams at intervals only", fullName="gatk_interval_file", shortName="intervals", required=false) var intervals: File = _ // Gracefully hide Queue's output val queueLogDir: String = ".qlog/" // Use the number of contigs for scatter gathering jobs var nContigs: Int = -1 // Updates and checks that all input files have the same number of contigs // we use the number of contigs for scatter gather. def updateNumberOfContigs(n: Int): Boolean = { if (nContigs < 0) { nContigs = n return true } return nContigs == n } def hasMultipleSamples(readGroups: java.util.List[SAMReadGroupRecord]): Boolean = { var sample: String = "" for (r <- readGroups) { if (sample.isEmpty) sample = r.getSample() else if (sample != r.getSample()) return true; } return false } def createSampleFiles(): Map[String, File] = { val outName: String = qscript.outputDir + qscript.projectName // Creating a table with SAMPLE information from each input BAM file val sampleTable = scala.collection.mutable.Map.empty[String, List[File]] for (bam <- scala.io.Source.fromFile(input).getLines) { val bamFile = new File(bam) val samReader = new SAMFileReader(bamFile) val header = samReader.getFileHeader() // keep a record of the number of contigs in this bam file (they should all be the same assert(updateNumberOfContigs(header.getSequenceDictionary.getSequences.size()), "Input BAMS should all have the same number of contigs. " + bam + " has " + header.getSequenceDictionary.getSequences.size()) val readGroups = header.getReadGroups() // only allow one sample per file. Bam files with multiple samples would require pre-processing of the file // with PrintReads to separate the samples. Tell user to do it himself! assert(!hasMultipleSamples(readGroups), "The pipeline requires that only one sample is present in a BAM file. Please separate the samples in " + bam) // Fill out the sample table with the readgroups in this file for (rg <- readGroups) { val sample = rg.getSample() if (!sampleTable.contains(sample)) sampleTable(sample) = List(bamFile) else if ( !sampleTable(sample).contains(bamFile)) sampleTable(sample) :+= bamFile } } // Creating one file for each sample in the dataset val sampleBamFiles = scala.collection.mutable.Map.empty[String, File] for ((sample, flist) <- sampleTable) { val sampleFileName = new File(outName + "." + sample + ".bam") sampleBamFiles(sample) = sampleFileName add(joinBams(flist, sampleFileName)) } return sampleBamFiles.toMap } def script = { //todo -- (option - BWA) run BWA on each bam file (per lane bam file) before performing per sample processing var cohortList: List[File] = List() val sampleBamFiles = createSampleFiles() // Simple progress report println("\nFound the following samples: ") for ((sample, file) <- sampleBamFiles) println("\t" + sample + " -> " + file) // If this is a 'knowns only' indel realignment run, do it only once for all samples. val globalIntervals = new File(outputDir + projectName + ".intervals") if (knownsOnly) add(target(null, globalIntervals)) // Put each sample through the pipeline for ((sample, bam) <- sampleBamFiles) { // BAM files generated by the pipeline val cleanedBam = swapExt(bam, ".bam", ".clean.bam") val dedupedBam = swapExt(bam, ".bam", ".clean.dedup.bam") val recalBam = swapExt(bam, ".bam", ".clean.dedup.recal.bam") // Accessory files val targetIntervals = if (knownsOnly) {globalIntervals} else {swapExt(bam, ".bam", ".intervals")} val metricsFile = swapExt(bam, ".bam", ".metrics") val preRecalFile = swapExt(bam, ".bam", ".pre_recal.csv") val postRecalFile = swapExt(bam, ".bam", ".post_recal.csv") val preOutPath = swapExt(bam, ".bam", ".pre") val postOutPath = swapExt(bam, ".bam", ".post") if (!knownsOnly) add(target(bam, targetIntervals)) add(clean(bam, targetIntervals, cleanedBam), dedup(cleanedBam, dedupedBam, metricsFile), cov(dedupedBam, preRecalFile), recal(dedupedBam, preRecalFile, recalBam), cov(recalBam, postRecalFile), analyzeCovariates(preRecalFile, preOutPath), analyzeCovariates(postRecalFile, postOutPath)) cohortList :+= recalBam } // output a BAM list with all the processed per sample files val cohortFile = new File(qscript.outputDir + qscript.projectName + ".cohort.list") add(writeList(cohortList, cohortFile)) } // General arguments to GATK walkers trait CommandLineGATKArgs extends CommandLineGATK { this.jarFile = qscript.GATKjar this.reference_sequence = qscript.reference this.memoryLimit = Some(4) this.isIntermediate = true } case class target (inBams: File, outIntervals: File) extends RealignerTargetCreator with CommandLineGATKArgs { if (!knownsOnly) this.input_file :+= inBams this.out = outIntervals this.mismatchFraction = Some(0.0) this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.rodBind :+= RodBind("indels", "VCF", indels) this.scatterCount = nContigs this.analysisName = queueLogDir + outIntervals + ".target" this.jobName = queueLogDir + outIntervals + ".target" } case class clean (inBams: File, tIntervals: File, outBam: File) extends IndelRealigner with CommandLineGATKArgs { this.input_file :+= inBams this.targetIntervals = tIntervals this.out = outBam this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.rodBind :+= RodBind("indels", "VCF", qscript.indels) this.useOnlyKnownIndels = knownsOnly this.doNotUseSW = useSW this.compress = Some(0) this.U = Some(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.scatterCount = nContigs this.analysisName = queueLogDir + outBam + ".clean" this.jobName = queueLogDir + outBam + ".clean" } //todo -- add scatter gather capability (waiting for khalid's modifications to the queue base case class cov (inBam: File, outRecalFile: File) extends CountCovariates with CommandLineGATKArgs { this.rodBind :+= RodBind("dbsnp", "VCF", dbSNP) this.covariate ++= List("ReadGroupCovariate", "QualityScoreCovariate", "CycleCovariate", "DinucCovariate") this.input_file :+= inBam this.recal_file = outRecalFile this.analysisName = queueLogDir + outRecalFile + ".covariates" this.jobName = queueLogDir + outRecalFile + ".covariates" } case class recal (inBam: File, inRecalFile: File, outBam: File) extends TableRecalibration with CommandLineGATKArgs { @Output(doc="recalibrated bam index") var recalIndex = new File(outBam + ".bai") this.input_file :+= inBam this.recal_file = inRecalFile this.baq = Some(org.broadinstitute.sting.utils.baq.BAQ.CalculationMode.CALCULATE_AS_NECESSARY) this.out = outBam if (!qscript.intervalString.isEmpty()) this.intervalsString ++= List(qscript.intervalString) else if (qscript.intervals != null) this.intervals :+= qscript.intervals this.U = Some(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 = Some(true) this.isIntermediate = false this.analysisName = queueLogDir + outBam + ".recalibration" this.jobName = queueLogDir + outBam + ".recalibration" } // Outside tools (not GATK walkers) case class analyzeCovariates (inRecalFile: File, outPath: File) extends AnalyzeCovariates { this.jarFile = qscript.ACJar this.resources = qscript.R this.recal_file = inRecalFile this.output_dir = outPath.toString this.analysisName = queueLogDir + inRecalFile + ".analyze_covariates" this.jobName = queueLogDir + inRecalFile + ".analyze_covariates" } case class dedup (inBam: File, outBam: File, metricsFile: File) extends PicardBamFunction { @Input(doc="fixed bam") var clean = inBam @Output(doc="deduped bam") var deduped = outBam @Output(doc="deduped bam index") var dedupedIndex = new File(outBam + ".bai") @Output(doc="metrics file") var metrics = metricsFile override def inputBams = List(clean) override def outputBam = deduped override def commandLine = super.commandLine + " M=" + metricsFile + " CREATE_INDEX=true" sortOrder = null this.memoryLimit = Some(6) this.isIntermediate = true this.jarFile = qscript.dedupJar this.analysisName = queueLogDir + outBam + ".dedup" this.jobName = queueLogDir + outBam + ".dedup" } case class joinBams (inBams: List[File], outBam: File) extends PicardBamFunction { @Input(doc="input bam list") var join = inBams @Output(doc="joined bam") var joined = outBam @Output(doc="joined bam index") var joinedIndex = new File(outBam + "bai") override def inputBams = join override def outputBam = joined override def commandLine = super.commandLine + " CREATE_INDEX=true" this.jarFile = qscript.mergeBamJar this.isIntermediate = true this.analysisName = queueLogDir + outBam + ".joinBams" this.jobName = queueLogDir + outBam + ".joinBams" } case class writeList(inBams: List[File], outBamList: File) extends ListWriterFunction { this.inputFiles = inBams this.listFile = outBamList this.analysisName = queueLogDir + outBamList + ".bamList" this.jobName = queueLogDir + outBamList + ".bamList" } }