/* * Copyright (c) 2009 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 org.broadinstitute.sting.gatk; import net.sf.picard.reference.ReferenceSequenceFile; import net.sf.picard.sam.SamFileHeaderMerger; import net.sf.picard.filter.SamRecordFilter; import net.sf.samtools.*; import org.apache.log4j.Logger; import org.broadinstitute.sting.gatk.datasources.simpleDataSources.SAMDataSource; import org.broadinstitute.sting.gatk.datasources.simpleDataSources.ReferenceOrderedDataSource; import org.broadinstitute.sting.gatk.datasources.shards.ShardStrategy; import org.broadinstitute.sting.gatk.datasources.shards.ShardStrategyFactory; import org.broadinstitute.sting.gatk.executive.MicroScheduler; import org.broadinstitute.sting.gatk.refdata.ReferenceOrderedData; import org.broadinstitute.sting.gatk.refdata.ReferenceOrderedDatum; import org.broadinstitute.sting.gatk.walkers.*; import org.broadinstitute.sting.gatk.filters.ZeroMappingQualityReadFilter; import org.broadinstitute.sting.gatk.io.OutputTracker; import org.broadinstitute.sting.utils.*; import org.broadinstitute.sting.utils.fasta.IndexedFastaSequenceFile; import org.broadinstitute.sting.utils.cmdLine.ArgumentException; import org.broadinstitute.sting.utils.cmdLine.ArgumentSource; import org.broadinstitute.sting.gatk.io.stubs.Stub; import java.io.File; import java.io.FileNotFoundException; import java.util.*; public class GenomeAnalysisEngine { // our instance of this genome analysis toolkit; it's used by other classes to extract the traversal engine // TODO: public static without final tends to indicate we're thinking about this the wrong way public static GenomeAnalysisEngine instance; /** * Accessor for sharded read data. */ private SAMDataSource readsDataSource = null; /** * Accessor for sharded reference data. */ private IndexedFastaSequenceFile referenceDataSource = null; /** * Accessor for sharded reference-ordered data. */ private List rodDataSources; // our argument collection private GATKArgumentCollection argCollection; /** Collection of inputs used by the walker. */ private Map inputs = new HashMap(); /** Collection of outputs used by the walker. */ private Collection> outputs = new ArrayList>(); /** our log, which we want to capture anything from this class */ private static Logger logger = Logger.getLogger(GenomeAnalysisEngine.class); /** our walker manager */ private final WalkerManager walkerManager; /** * our constructor, where all the work is done *

* legacy traversal types are sent to legacyTraversal function; as we move more of the traversals to the * new MicroScheduler class we'll be able to delete that function. * */ public GenomeAnalysisEngine() { // make sure our instance variable points to this analysis engine instance = this; walkerManager = new WalkerManager(); } /** * Actually run the GATK with the specified walker. * @param args the argument collection, where we get all our setup information from * @param my_walker Walker to run over the dataset. Must not be null. */ public Object execute(GATKArgumentCollection args, Walker my_walker) { // validate our parameters if (args == null) { throw new StingException("The GATKArgumentCollection passed to GenomeAnalysisEngine can not be null."); } // validate our parameters if (my_walker == null) throw new StingException("The walker passed to GenomeAnalysisEngine can not be null."); // save our argument parameter this.argCollection = args; // Prepare the data for traversal. initializeDataSources( my_walker, argCollection ); // our microscheduler, which is in charge of running everything MicroScheduler microScheduler = createMicroscheduler(my_walker); // create the output streams initializeOutputStreams(my_walker, microScheduler.getOutputTracker()); GenomeLocSortedSet locs = null; if (argCollection.intervals != null) { locs = GenomeLocSortedSet.createSetFromList(parseIntervalRegion(argCollection.intervals)); } ShardStrategy shardStrategy = getShardStrategy(my_walker, microScheduler.getReference(), locs, argCollection.maximumEngineIterations); // execute the microscheduler, storing the results return microScheduler.execute(my_walker, shardStrategy, argCollection.maximumEngineIterations); } /** * Add additional, externally managed IO streams for walker input. * @param argumentSource Field in the walker into which to inject the value. * @param value Instance to inject. */ public void addInput( ArgumentSource argumentSource, Object value ) { inputs.put(argumentSource,value); } /** * Add additional, externally managed IO streams for walker output. * @param stub Instance to inject. */ public void addOutput( Stub stub ) { outputs.add(stub); } /** * Gets a set of the names of all walkers that the GATK has discovered. * @return A set of the names of all discovered walkers. */ public Set getWalkerNames() { return walkerManager.getWalkerNames(); } /** * Retrieves an instance of the walker based on the walker name. * @param walkerName Name of the walker. Must not be null. If the walker cannot be instantiated, an exception will be thrown. * @return An instance of the walker. */ public Walker getWalkerByName( String walkerName ) { return walkerManager.createWalkerByName(walkerName); } private void initializeDataSources( Walker my_walker, GATKArgumentCollection argCollection ) { validateSuppliedReadsAgainstWalker( my_walker, argCollection ); logger.info("Strictness is " + argCollection.strictnessLevel); readsDataSource = createReadsDataSource(extractSourceInfo(my_walker,argCollection)); validateSuppliedReferenceAgainstWalker( my_walker, argCollection ); referenceDataSource = openReferenceSequenceFile(argCollection.referenceFile); validateReadsAndReferenceAreCompatible(readsDataSource, referenceDataSource); // our reference ordered data collection List> rods = new ArrayList>(); // // please don't use these in the future, use the new syntax <- if we're not using these please remove them // if (argCollection.DBSNPFile != null) bindConvenienceRods("dbSNP", "dbsnp", argCollection.DBSNPFile); if (argCollection.HAPMAPFile != null) bindConvenienceRods("hapmap", "HapMapAlleleFrequencies", argCollection.HAPMAPFile); if (argCollection.HAPMAPChipFile != null) bindConvenienceRods("hapmap-chip", "GFF", argCollection.HAPMAPChipFile); // TODO: The ROD iterator currently does not understand multiple intervals file. Fix this by cleaning the ROD system. if (argCollection.intervals != null && argCollection.intervals.size() == 1) { bindConvenienceRods("interval", "Intervals", argCollection.intervals.get(0).replaceAll(",", "")); } // parse out the rod bindings ReferenceOrderedData.parseBindings(argCollection.RODBindings, rods); validateSuppliedReferenceOrderedDataAgainstWalker( my_walker, rods ); rodDataSources = getReferenceOrderedDataSources(rods); } /** * setup a microscheduler * @param my_walker our walker of type LocusWalker * @return a new microscheduler */ private MicroScheduler createMicroscheduler(Walker my_walker) { // the mircoscheduler to return MicroScheduler microScheduler = null; // we need to verify different parameter based on the walker type if (my_walker instanceof LocusWalker || my_walker instanceof LocusWindowWalker) { // create the MicroScheduler microScheduler = MicroScheduler.create(my_walker, readsDataSource, referenceDataSource, rodDataSources, argCollection.numberOfThreads); } else if (my_walker instanceof ReadWalker || my_walker instanceof DuplicateWalker) { if (argCollection.referenceFile == null) Utils.scareUser(String.format("Read-based traversals require a reference file but none was given")); microScheduler = MicroScheduler.create(my_walker, readsDataSource, referenceDataSource, rodDataSources, argCollection.numberOfThreads); } else { Utils.scareUser(String.format("Unable to create the appropriate TraversalEngine for analysis type " + argCollection.analysisName)); } return microScheduler; } /** * setup the interval regions, from either the interval file of the genome region string * * @param intervals the list of intervals to parse * * @return a list of genomeLoc representing the interval file */ public static List parseIntervalRegion(final List intervals) { List locs = new ArrayList(); for (String interval : intervals) { if (new File(interval).exists()) { locs.addAll(GenomeLocParser.intervalFileToList(interval)); } else { locs.addAll(GenomeLocParser.parseGenomeLocs(interval)); } } return locs; } /** * Returns sets of samples present in the (merged) input SAM stream, grouped by readers (i.e. underlying * individual bam files). For instance: if GATK is run with three input bam files (three -I arguments), then the list * returned by this method will contain 3 elements (one for each reader), with each element being a set of sample names * found in the corresponding bam file. * @return */ public List< Set > getSamplesByReaders() { SamFileHeaderMerger hm = getDataSource().getHeaderMerger(); List< Set > sample_sets = new ArrayList>(hm.getReaders().size()); for ( SAMFileReader r : hm.getReaders() ) { Set samples = new HashSet(1); sample_sets.add(samples); for ( SAMReadGroupRecord g : r.getFileHeader().getReadGroups() ) { samples.add(g.getSample()); } } return sample_sets; } /** * Returns sets of libraries present in the (merged) input SAM stream, grouped by readers (i.e. underlying * individual bam files). For instance: if GATK is run with three input bam files (three -I arguments), then the list * returned by this method will contain 3 elements (one for each reader), with each element being a set of library names * found in the corresponding bam file. * @return */ public List< Set > getLibrariesByReaders() { SamFileHeaderMerger hm = getDataSource().getHeaderMerger(); List< Set > lib_sets = new ArrayList>(hm.getReaders().size()); for ( SAMFileReader r : hm.getReaders() ) { Set libs = new HashSet(2); lib_sets.add(libs); for ( SAMReadGroupRecord g : r.getFileHeader().getReadGroups() ) { libs.add(g.getLibrary()); } } return lib_sets; } /** * Returns sets of (remapped) read groups in input SAM stream, grouped by readers (i.e. underlying * individual bam files). For instance: if GATK is run with three input bam files (three -I arguments), then the list * returned by this method will contain 3 elements (one for each reader), with each element being a set of remapped read groups * (i.e. as seen by read.getReadGroup().getReadGroupId() in the merged stream) that come from the corresponding bam file. * @return */ public List< Set > getMergedReadGroupsByReaders() { SamFileHeaderMerger hm = getDataSource().getHeaderMerger(); List< Set > rg_sets = new ArrayList>(hm.getReaders().size()); for ( SAMFileReader r : hm.getReaders() ) { Set groups = new HashSet(5); rg_sets.add(groups); for ( SAMReadGroupRecord g : r.getFileHeader().getReadGroups() ) { // use HeaderMerger to translate original read group id from the reader into the read group id in the // merged stream, and save that remapped read group id to associate it with specific reader groups.add( hm.getReadGroupId(r, g.getReadGroupId()) ); } } return rg_sets; } /** * Bundles all the source information about the reads into a unified data structure. * * @param walker The walker for which to extract info. * @param argCollection The collection of arguments passed to the engine. * * @return The reads object providing reads source info. */ private Reads extractSourceInfo( Walker walker, GATKArgumentCollection argCollection ) { List filters = new ArrayList(); filters.addAll( WalkerManager.getReadFilters(walker) ); if( argCollection.filterZeroMappingQualityReads != null && argCollection.filterZeroMappingQualityReads ) filters.add( new ZeroMappingQualityReadFilter() ); return new Reads( argCollection.samFiles, argCollection.strictnessLevel, argCollection.downsampleFraction, argCollection.downsampleCoverage, !argCollection.unsafe, filters, argCollection.readMaxPileup); } /** * Verifies that the supplied set of reads files mesh with what the walker says it requires. * @param walker Walker to test. * @param arguments Supplied reads files. */ private void validateSuppliedReadsAgainstWalker( Walker walker, GATKArgumentCollection arguments ) { // Check what the walker says is required against what was provided on the command line. if (WalkerManager.isRequired(walker, DataSource.READS) && (arguments.samFiles == null || arguments.samFiles.size() == 0)) throw new ArgumentException("Walker requires reads but none were provided. If this is incorrect, alter the walker's @Requires annotation."); // Check what the walker says is allowed against what was provided on the command line. if ((arguments.samFiles != null && arguments.samFiles.size() > 0) && !WalkerManager.isAllowed(walker, DataSource.READS)) throw new ArgumentException("Walker does not allow reads but reads were provided. If this is incorrect, alter the walker's @Allows annotation"); } /** * Verifies that the supplied reference file mesh with what the walker says it requires. * @param walker Walker to test. * @param arguments Supplied reads files. */ private void validateSuppliedReferenceAgainstWalker( Walker walker, GATKArgumentCollection arguments ) { // Check what the walker says is required against what was provided on the command line. if (WalkerManager.isRequired(walker, DataSource.REFERENCE) && arguments.referenceFile == null) throw new ArgumentException("Walker requires a reference but none was provided. If this is incorrect, alter the walker's @Requires annotation."); // Check what the walker says is allowed against what was provided on the command line. if (arguments.referenceFile != null && !WalkerManager.isAllowed(walker, DataSource.REFERENCE)) throw new ArgumentException("Walker does not allow a reference but one was provided. If this is incorrect, alter the walker's @Allows annotation"); } /** * Verifies that all required reference-ordered data has been supplied, and any reference-ordered data that was not * 'allowed' is still present. * @param walker Walker to test. * @param rods Reference-ordered data to load. */ private void validateSuppliedReferenceOrderedDataAgainstWalker( Walker walker, List> rods ) { // Check to make sure that all required metadata is present. List allRequired = WalkerManager.getRequiredMetaData(walker); for (RMD required : allRequired) { boolean found = false; for (ReferenceOrderedData rod : rods) { if (rod.matches(required.name(), required.type())) found = true; } if (!found) throw new ArgumentException(String.format("Unable to find reference metadata (%s,%s)", required.name(), required.type())); } // Check to see that no forbidden rods are present. for (ReferenceOrderedData rod : rods) { if (!WalkerManager.isAllowed(walker, rod)) throw new ArgumentException(String.format("Walker does not allow access to metadata: %s. If this is incorrect, change the @Allows metadata", rod.getName())); } } /** * Now that all files are open, validate the sequence dictionaries of the reads vs. the reference. * @param reads Reads data source. * @param reference Reference data source. */ private void validateReadsAndReferenceAreCompatible( SAMDataSource reads, ReferenceSequenceFile reference ) { if( reads == null || reference == null ) return; // Compile a set of sequence names that exist in the BAM files. SAMSequenceDictionary readsDictionary = reads.getHeader().getSequenceDictionary(); Set readsSequenceNames = new TreeSet(); for( SAMSequenceRecord dictionaryEntry: readsDictionary.getSequences() ) readsSequenceNames.add(dictionaryEntry.getSequenceName()); // Compile a set of sequence names that exist in the reference file. SAMSequenceDictionary referenceDictionary = reference.getSequenceDictionary(); Set referenceSequenceNames = new TreeSet(); for( SAMSequenceRecord dictionaryEntry: referenceDictionary.getSequences() ) referenceSequenceNames.add(dictionaryEntry.getSequenceName()); if( readsSequenceNames.size() == 0 ) { logger.info("Reads file is unmapped. Skipping validation against reference."); return; } // If there's no overlap between reads and reference, data will be bogus. Throw an exception. Set intersectingSequenceNames = new HashSet(readsSequenceNames); intersectingSequenceNames.retainAll(referenceSequenceNames); if( intersectingSequenceNames.size() == 0 ) { StringBuilder error = new StringBuilder(); error.append("No overlap exists between sequence dictionary of the reads and the sequence dictionary of the reference. Perhaps you're using the wrong reference?\n"); error.append(System.getProperty("line.separator")); error.append(String.format("Reads contigs: %s%n", prettyPrintSequenceRecords(readsDictionary))); error.append(String.format("Reference contigs: %s%n", prettyPrintSequenceRecords(referenceDictionary))); logger.error(error.toString()); Utils.scareUser("No overlap exists between sequence dictionary of the reads and the sequence dictionary of the reference."); } // If the two datasets are not equal and neither is a strict subset of the other, warn the user. if( !readsSequenceNames.equals(referenceSequenceNames) && !readsSequenceNames.containsAll(referenceSequenceNames) && !referenceSequenceNames.containsAll(readsSequenceNames)) { StringBuilder warning = new StringBuilder(); warning.append("Limited overlap exists between sequence dictionary of the reads and the sequence dictionary of the reference. Perhaps you're using the wrong reference?\n"); warning.append(System.getProperty("line.separator")); warning.append(String.format("Reads contigs: %s%n", prettyPrintSequenceRecords(readsDictionary))); warning.append(String.format("Reference contigs: %s%n", prettyPrintSequenceRecords(referenceDictionary))); logger.warn(warning.toString()); } } private String prettyPrintSequenceRecords( SAMSequenceDictionary sequenceDictionary ) { String[] sequenceRecordNames = new String[ sequenceDictionary.size() ]; int sequenceRecordIndex = 0; for( SAMSequenceRecord sequenceRecord: sequenceDictionary.getSequences() ) sequenceRecordNames[sequenceRecordIndex++] = sequenceRecord.getSequenceName(); return Arrays.deepToString(sequenceRecordNames); } /** * Convenience function that binds RODs using the old-style command line parser to the new style list for * a uniform processing. * * @param name the name of the rod * @param type its type * @param file the file to load the rod from */ private void bindConvenienceRods(final String name, final String type, final String file) { argCollection.RODBindings.add(Utils.join(",", new String[]{name, type, file})); } /** * Get the sharding strategy given a driving data source. * * @param walker Walker for which to infer sharding strategy. * @param drivingDataSource Data on which to shard. * @param intervals Intervals to use when limiting sharding. * @param maxIterations the maximum number of iterations to run through * * @return Sharding strategy for this driving data source. */ protected ShardStrategy getShardStrategy(Walker walker, ReferenceSequenceFile drivingDataSource, GenomeLocSortedSet intervals, Integer maxIterations) { final long SHARD_SIZE = 100000L; ShardStrategy shardStrategy = null; ShardStrategyFactory.SHATTER_STRATEGY shardType; if (walker instanceof LocusWalker) { if (intervals != null) { shardType = (walker.isReduceByInterval()) ? ShardStrategyFactory.SHATTER_STRATEGY.INTERVAL : ShardStrategyFactory.SHATTER_STRATEGY.LINEAR; shardStrategy = ShardStrategyFactory.shatter(shardType, drivingDataSource.getSequenceDictionary(), SHARD_SIZE, intervals, maxIterations); } else shardStrategy = ShardStrategyFactory.shatter(ShardStrategyFactory.SHATTER_STRATEGY.LINEAR, drivingDataSource.getSequenceDictionary(), SHARD_SIZE, maxIterations); } else if (walker instanceof ReadWalker || walker instanceof DuplicateWalker) { shardType = ShardStrategyFactory.SHATTER_STRATEGY.READS; if (intervals != null) { shardStrategy = ShardStrategyFactory.shatter(shardType, drivingDataSource.getSequenceDictionary(), SHARD_SIZE, intervals, maxIterations); } else { shardStrategy = ShardStrategyFactory.shatter(shardType, drivingDataSource.getSequenceDictionary(), SHARD_SIZE, maxIterations); } } else if (walker instanceof LocusWindowWalker) { if( intervals == null ) throw new StingException("Unable to shard: walker is of type LocusWindow, but no intervals were provided"); shardStrategy = ShardStrategyFactory.shatter(ShardStrategyFactory.SHATTER_STRATEGY.INTERVAL, drivingDataSource.getSequenceDictionary(), SHARD_SIZE, intervals, maxIterations); } else throw new StingException("Unable to support walker of type" + walker.getClass().getName()); return shardStrategy; } /** * Gets a data source for the given set of reads. * * @param reads the read source information * * @return A data source for the given set of reads. */ private SAMDataSource createReadsDataSource(Reads reads) { // By reference traversals are happy with no reads. Make sure that case is handled. if (reads.getReadsFiles().size() == 0) return null; SAMDataSource dataSource = new SAMDataSource(reads); return dataSource; } /** * Opens a reference sequence file paired with an index. * * @param refFile Handle to a reference sequence file. Non-null. * * @return A thread-safe file wrapper. */ private IndexedFastaSequenceFile openReferenceSequenceFile(File refFile) { IndexedFastaSequenceFile ref = null; try { ref = new IndexedFastaSequenceFile(refFile); } catch (FileNotFoundException ex) { throw new StingException("I/O error while opening fasta file: " + ex.getMessage(), ex); } GenomeLocParser.setupRefContigOrdering(ref); return ref; } /** * Open the reference-ordered data sources. * * @param rods the reference order data to execute using * * @return A list of reference-ordered data sources. */ private List getReferenceOrderedDataSources(List> rods) { List dataSources = new ArrayList(); for (ReferenceOrderedData rod : rods) dataSources.add(new ReferenceOrderedDataSource(rod)); return dataSources; } /** * Initialize the output streams as specified by the user. * * @param walker the walker to initialize output streams for */ private void initializeOutputStreams(Walker walker, OutputTracker outputTracker) { if( argCollection.outErrFileName != null ) outputTracker.initializeCoreIO( argCollection.outErrFileName, argCollection.outErrFileName ); else outputTracker.initializeCoreIO( argCollection.outFileName, argCollection.errFileName ); for( Map.Entry input: inputs.entrySet() ) outputTracker.addInput(input.getKey(),input.getValue()); for( Stub stub: outputs ) outputTracker.addOutput(stub); outputTracker.prepareWalker(walker); } public SAMFileHeader getSAMFileHeader() { return readsDataSource.getHeader(); } /** * Returns data source object encapsulating all essential info and handlers used to traverse * reads; header merger, individual file readers etc can be accessed through the returned data source object. * @return */ public SAMDataSource getDataSource() { return this.readsDataSource; } /** * Gets the collection of GATK main application arguments for enhanced walker validation. * * @return the GATK argument collection */ public GATKArgumentCollection getArguments() { return this.argCollection; } }