/* * The MIT License * * Copyright (c) 2014 The Broad Institute * * Permission is hereby granted, free of charge, to any person obtaining a copy * of this software and associated documentation files (the "Software"), to deal * in the Software without restriction, including without limitation the rights * to use, copy, modify, merge, publish, distribute, sublicense, and/or sell * copies of the Software, and to permit persons to whom the Software is * furnished to do so, subject to the following conditions: * * The above copyright notice and this permission notice shall be included in * all copies or substantial portions of the Software. * * THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR * IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, * FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE * AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER * LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, * OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN * THE SOFTWARE. */ package picard.sam.markduplicates; import picard.cmdline.CommandLineProgramProperties; import picard.cmdline.Option; import htsjdk.samtools.util.Histogram; import htsjdk.samtools.util.IterableAdapter; import htsjdk.samtools.util.Log; import htsjdk.samtools.util.IOUtil; import htsjdk.samtools.util.ProgressLogger; import htsjdk.samtools.*; import picard.cmdline.programgroups.SamOrBam; import picard.sam.markduplicates.util.AbstractMarkDuplicatesCommandLineProgram; import java.util.*; /** * An even better duplication marking algorithm that handles all cases including clipped * and gapped alignments. * <p/> * This tool differs with MarkDuplicates as it may break ties differently. Furthermore, * as it is a one-pass algorithm, it cannot know the program records contained in the file * that should be chained in advance. Therefore it will only be able to examine the header * to attempt to infer those program group records that have no associated previous program * group record. If a read is encountered without a program record, or not one as previously * defined, it will not be updated. * <p/> * This tool will also not work with alignments that have large gaps or skips, such as those * from RNA-seq data. This is due to the need to buffer small genomic windows to ensure * integrity of the duplicate marking, while large skips (ex. skipping introns) in the * alignment records would force making that window very large, thus exhausting memory. * * @author Nils Homer */ @CommandLineProgramProperties( usage = "Examines aligned records in the supplied SAM or BAM file to locate duplicate molecules. " + "All records are then written to the output file with the duplicate records flagged.", usageShort = "Examines aligned records in the supplied SAM or BAM file to locate duplicate molecules.", programGroup = SamOrBam.class ) public class MarkDuplicatesWithMateCigar extends AbstractMarkDuplicatesCommandLineProgram { private final Log log = Log.getInstance(MarkDuplicatesWithMateCigar.class); @Option(doc = "The minimum distance to buffer records to account for clipping on the 5' end of the records." + "Set this number to -1 to use twice the first read's read length (or 100, whichever is smaller).", optional = true) public int MINIMUM_DISTANCE = -1; @Option(doc = "Skip record pairs with no mate cigar and include them in the output.") boolean SKIP_PAIRS_WITH_NO_MATE_CIGAR = true; @Option(doc = "The block size for use in the coordinate-sorted record buffer.", optional = true) public int BLOCK_SIZE = 100000; /** Warnings that will only be emitted once */ private boolean warnedNullProgramRecords = false; private boolean warnedMissingProgramRecords = false; /** Stock main method. */ public static void main(final String[] args) { new MarkDuplicatesWithMateCigar().instanceMainWithExit(args); } /** * Main work method. */ protected int doWork() { IOUtil.assertFilesAreReadable(INPUT); IOUtil.assertFileIsWritable(OUTPUT); IOUtil.assertFileIsWritable(METRICS_FILE); // Open the inputs final SamHeaderAndIterator headerAndIterator = openInputs(); final SAMFileHeader header = headerAndIterator.header; // Create the output header final SAMFileHeader outputHeader = header.clone(); outputHeader.setSortOrder(SAMFileHeader.SortOrder.coordinate); for (final String comment : COMMENT) outputHeader.addComment(comment); // Since this is one-pass, unlike MarkDuplicates, we cannot only chain together program // group records we have seen, we have to assume all of them may be seen. We can perhaps // filter out any program groups which have been referenced previously. setPGIdsSeen(outputHeader); // Key: previous PG ID on a SAM Record (or null). Value: New PG ID to replace it. final Map<String, String> chainedPgIds = getChainedPgIds(outputHeader); // Open the output final SAMFileWriter out = new SAMFileWriterFactory().makeSAMOrBAMWriter(outputHeader, true, OUTPUT); // Create the mark duplicate iterator. The duplicate marking is handled by the iterator, conveniently. final MarkDuplicatesWithMateCigarIterator iterator = new MarkDuplicatesWithMateCigarIterator(headerAndIterator.header, headerAndIterator.iterator, this.opticalDuplicateFinder, this.DUPLICATE_SCORING_STRATEGY, this.MINIMUM_DISTANCE, this.REMOVE_DUPLICATES, this.SKIP_PAIRS_WITH_NO_MATE_CIGAR, this.MAX_RECORDS_IN_RAM, this.BLOCK_SIZE, this.TMP_DIR); // progress logger! final ProgressLogger progress = new ProgressLogger(log, (int) 1e6, "Read"); // Go through the records for (final SAMRecord record : new IterableAdapter<SAMRecord>(iterator)) { if (progress.record(record)) { iterator.logMemoryStats(log); } // Update the program record if necessary updateProgramRecord(record, chainedPgIds); // Write the alignment out.addAlignment(record); } // remember to close the inputs iterator.close(); out.close(); // For convenience to reference final Histogram<Short> opticalDupesByLibraryId = iterator.getOpticalDupesByLibraryId(); // Log info log.info("Processed " + progress.getCount() + " records"); log.info("Found " + iterator.getNumRecordsWithNoMateCigar() + " records with no mate cigar optional tag."); log.info("Marking " + iterator.getNumDuplicates() + " records as duplicates."); log.info("Found " + ((long) opticalDupesByLibraryId.getSumOfValues()) + " optical duplicate clusters."); // cast as long due to returning a double // Write out the metrics finalizeAndWriteMetrics(iterator.getLibraryIdGenerator()); return 0; } /** * Updates the program record if necessary. */ private void updateProgramRecord(final SAMRecord record, final Map<String, String> chainedPgIds) { if (PROGRAM_RECORD_ID != null) { final String pgId = record.getStringAttribute(SAMTag.PG.name()); if (null == pgId) { if (!warnedNullProgramRecords) { warnedNullProgramRecords = true; log.warn("Encountered a record with no program record, program group chaining will not occur for this read: " + record); } // else already warned! } else if (!chainedPgIds.containsKey(pgId)) { if (!warnedMissingProgramRecords) { warnedMissingProgramRecords = true; log.warn("Encountered a record with an intermediate program record, program group chaining will not occur for this read: " + record); } // else already warned! } else { record.setAttribute(SAMTag.PG.name(), chainedPgIds.get(pgId)); } } } /** * Generate the list of program records seen in the SAM file, approximating this with those in the header that were not * themselves mentioned elsewhere. */ private void setPGIdsSeen(final SAMFileHeader header) { final Set<String> pgIdsSeenAsPrevious = new HashSet<String>(); // get all program record ids that are mentioned as previously seen for (final SAMProgramRecord samProgramRecord : header.getProgramRecords()) { final String previousProgramGroupID = samProgramRecord.getPreviousProgramGroupId(); if (null != previousProgramGroupID) pgIdsSeenAsPrevious.add(previousProgramGroupID); } // ignore those that were previously seen for (final SAMProgramRecord samProgramRecord : header.getProgramRecords()) { final String pgId = samProgramRecord.getId(); if (!pgIdsSeenAsPrevious.contains(pgId)) this.pgIdsSeen.add(pgId); } } }