package org.broadinstitute.sting.gatk; import net.sf.picard.reference.ReferenceSequenceFile; import net.sf.picard.reference.ReferenceSequenceFileFactory; import net.sf.samtools.SAMFileReader; import net.sf.samtools.SAMFileReader.ValidationStringency; import org.apache.log4j.Logger; 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.refdata.RODIterator; import org.broadinstitute.sting.gatk.refdata.IntervalRodIterator; import org.broadinstitute.sting.gatk.traversals.*; import org.broadinstitute.sting.gatk.walkers.*; import org.broadinstitute.sting.utils.*; import org.broadinstitute.sting.utils.cmdLine.ArgumentException; import java.util.ArrayList; import java.util.List; import java.io.File; 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; // our traversal engine private TraversalEngine engine = null; // the level of debugging we're using public boolean DEBUGGING = false; // our argument collection private final GATKArgumentCollection argCollection; /** Collection of output streams used by the walker. */ private OutputTracker outputTracker = null; /** our log, which we want to capture anything from this class */ private static Logger logger = Logger.getLogger(GenomeAnalysisEngine.class); /** the return value from our walker */ private Object walkerReturn = null; /** * 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. * * @param args the argument collection, where we get all our setup information from * @param my_walker the walker we're running with */ public GenomeAnalysisEngine(GATKArgumentCollection args, Walker my_walker) { // validate our parameters if (args == null || my_walker == null) { throw new StingException("Neither the GATKArgumentCollection or the Walker passed to GenomeAnalysisEngine can be null."); } // save our argument parameter this.argCollection = args; // make sure our instance variable points to this analysis engine instance = this; // 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); if ( argCollection.intervals != null ) bindConvenienceRods("interval", "Intervals", argCollection.intervals.replaceAll(",", "")); // parse out the rod bindings ReferenceOrderedData.parseBindings(logger, argCollection.RODBindings, rods); // Validate the walker inputs against the walker. validateInputsAgainstWalker(my_walker, argCollection, rods); // create the output streams initializeOutputStreams( my_walker ); // our microscheduler, which is in charge of running everything MicroScheduler microScheduler = null; // if we're a read or a locus walker, we use the new system. Right now we have complicated // branching based on the input data, but this should disapear when all the traversals are switched over if (my_walker instanceof LocusWalker || my_walker instanceof ReadWalker) { microScheduler = createMicroscheduler(my_walker, rods); } else { // we have an old style traversal, once we're done return legacyTraversal(my_walker, rods); return; } // Prepare the sort ordering w.r.t. the sequence dictionary if (argCollection.referenceFile != null) { final ReferenceSequenceFile refFile = ReferenceSequenceFileFactory.getReferenceSequenceFile(argCollection.referenceFile); GenomeLoc.setupRefContigOrdering(refFile); } // Determine the validation stringency. Default to ValidationStringency.STRICT. ValidationStringency strictness = getValidationStringency(); logger.info("Strictness is " + strictness); // perform validation steps that are common to all the engines genericEngineSetup(strictness); // parse out any genomic location they've provided //List locationsList = setupIntervalRegion(); List locationsList = engine.getLocations(); GenomeLocSortedSet locs = null; if (locationsList != null) locs = GenomeLocSortedSet.createSetFromList(locationsList); // excute the microscheduler, storing the results walkerReturn = microScheduler.execute(my_walker, locs, argCollection.maximumEngineIterations); } /** * this is to accomdate the older style traversals, that haven't been converted over to the new system. Putting them * into their own function allows us to deviate in the two behaviors so the new style traversals aren't limited to what * the old style does. As traversals are converted, this function should disappear. * * @param my_walker * @param rods */ private void legacyTraversal(Walker my_walker, List> rods) { if (my_walker instanceof LocusWindowWalker) { this.engine = new TraverseByLocusWindows(argCollection.samFiles, argCollection.referenceFile, rods); } else if (my_walker instanceof DuplicateWalker) { // we're a duplicate walker this.engine = new TraverseDuplicates(argCollection.samFiles, argCollection.referenceFile, rods); } else { throw new RuntimeException("Unexpected walker type: " + my_walker); } // Prepare the sort ordering w.r.t. the sequence dictionary if (argCollection.referenceFile != null) { final ReferenceSequenceFile refFile = ReferenceSequenceFileFactory.getReferenceSequenceFile(argCollection.referenceFile); GenomeLoc.setupRefContigOrdering(refFile); } // Determine the validation stringency. Default to ValidationStringency.STRICT. ValidationStringency strictness = getValidationStringency(); logger.info("Strictness is " + strictness); genericEngineSetup(strictness); // store the results of the walker run walkerReturn = engine.traverse(my_walker); } /** * setup a microscheduler * * @param my_walker our walker of type LocusWalker * @param rods the reference order data * @return a new microscheduler */ private MicroScheduler createMicroscheduler(Walker my_walker, List> rods) { // the mircoscheduler to return MicroScheduler microScheduler = null; // we need to verify different parameter based on the walker type if (my_walker instanceof LocusWalker) { // create the MicroScheduler if( argCollection.walkAllLoci ) Utils.scareUser("Argument --all_loci is deprecated. Please annotate your walker with @By(DataSource.REFERENCE) to perform a by-reference traversal."); microScheduler = MicroScheduler.create(my_walker, new Reads(argCollection), argCollection.referenceFile, rods, argCollection.numberOfThreads); engine = microScheduler.getTraversalEngine(); } else if (my_walker instanceof ReadWalker) { if (argCollection.referenceFile == null) Utils.scareUser(String.format("Locus-based traversals require a reference file but none was given")); microScheduler = MicroScheduler.create(my_walker, new Reads(argCollection), argCollection.referenceFile, rods, argCollection.numberOfThreads); engine = microScheduler.getTraversalEngine(); } return microScheduler; } /** * commands that get executed for each engine, regardless of the type * * @param strictness our current strictness level */ private void genericEngineSetup(ValidationStringency strictness) { engine.setStrictness(strictness); engine.setMaxReads(argCollection.maximumEngineIterations); engine.setFilterZeroMappingQualityReads(argCollection.filterZeroMappingQualityReads); // we default interval files over the genome region string if (argCollection.intervals != null) { engine.setLocation(parseIntervalRegion(argCollection.intervals, false)); } engine.setReadFilters(new Reads(argCollection)); engine.setThreadedIO(argCollection.enabledThreadedIO); engine.setWalkOverAllSites(argCollection.walkAllLoci); engine.initialize(); } /** * setup the interval regions, from either the interval file of the genome region string * * @return a list of genomeLoc representing the interval file */ public static List parseIntervalRegion(final String intervalsString, boolean quiet ) { List locs = null; if ( intervalsString != null) { if (new File(intervalsString).exists()) { if (! quiet) logger.info("Intervals argument specifies a file. Loading intervals from file."); locs = GenomeLoc.IntervalFileToList(intervalsString); } else { if (! quiet) logger.info("Intervals argument does not specify a file. Trying to parse it as a simple string."); locs = GenomeLoc.parseGenomeLocs(intervalsString); } } return locs; } private void validateInputsAgainstWalker(Walker walker, GATKArgumentCollection arguments, List> rods) { String walkerName = WalkerManager.getWalkerName(walker.getClass()); // 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(String.format("Walker %s requires reads but none were provided. If this is incorrect, alter the walker's @Requires annotation.",walkerName)); if( WalkerManager.isRequired(walker,DataSource.REFERENCE) && arguments.referenceFile == null ) throw new ArgumentException(String.format("Walker %s requires a reference but none was provided. If this is incorrect, alter the walker's @Requires annotation.",walkerName)); // 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(String.format("Walker %s does not allow reads but reads were provided. If this is incorrect, alter the walker's @Allows annotation",walkerName)); if( arguments.referenceFile != null && !WalkerManager.isAllowed(walker,DataSource.REFERENCE) ) throw new ArgumentException(String.format("Walker %s does not allow a reference but one was provided. If this is incorrect, alter the walker's @Allows annotation",walkerName)); // 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 correct, change the @Allows metadata",rod.getName())); } } /** * Default to ValidationStringency.STRICT. * * @return the validation stringency */ private ValidationStringency getValidationStringency() { ValidationStringency strictness; try { strictness = Enum.valueOf(ValidationStringency.class, argCollection.strictnessLevel); } catch (IllegalArgumentException ex) { strictness = ValidationStringency.STRICT; } return strictness; } /** * Default to 5 (based on research by Alec Wysoker) * * @return the BAM compression */ public int getBAMCompression() { return (argCollection.BAMcompression == null || argCollection.BAMcompression < 1 || argCollection.BAMcompression > 8) ? 5 : argCollection.BAMcompression; } /** * Convenience function that binds RODs using the old-style command line parser to the new style list for * a uniform processing. * * @param name * @param type * @param file */ private void bindConvenienceRods(final String name, final String type, final String file) { argCollection.RODBindings.add(Utils.join(",", new String[]{name, type, file})); } /** Initialize the output streams as specified by the user. */ private void initializeOutputStreams( Walker walker ) { outputTracker = (argCollection.outErrFileName != null) ? new OutputTracker(argCollection.outErrFileName, argCollection.outErrFileName) : new OutputTracker(argCollection.outFileName, argCollection.errFileName); walker.initializeOutputStreams(outputTracker); } /** * Gets the output tracker. Tracks data available to a given walker. * * @return The output tracker. */ public OutputTracker getOutputTracker() { return outputTracker; } /** * This function is deprecated in the new traversal engines, if you need to get the header * please get the engine and then use the getHeader function * * @return */ @Deprecated public SAMFileReader getSamReader() { return this.engine.getSamReader(); } public TraversalEngine getEngine() { return this.engine; } /** Gets the collection of GATK main application arguments for enhanced walker validation. */ public GATKArgumentCollection getArguments() { return this.argCollection; } /** * Get's the return value of the walker * * @return an Object representing the return value of the walker */ public Object getWalkerReturn() { return walkerReturn; } }