477 lines
21 KiB
Java
477 lines
21 KiB
Java
package org.broadinstitute.sting.gatk;
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import edu.mit.broad.picard.reference.ReferenceSequence;
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import edu.mit.broad.picard.reference.ReferenceSequenceFile;
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import edu.mit.broad.picard.reference.ReferenceSequenceFileFactory;
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import net.sf.samtools.SAMFileReader;
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import net.sf.samtools.SAMFileReader.ValidationStringency;
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import net.sf.samtools.SAMSequenceRecord;
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import net.sf.samtools.util.RuntimeIOException;
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import org.apache.log4j.Logger;
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import org.apache.commons.cli.OptionBuilder;
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import org.apache.commons.cli.Option;
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import org.broadinstitute.sting.gatk.refdata.ReferenceOrderedData;
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import org.broadinstitute.sting.gatk.refdata.ReferenceOrderedDatum;
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import org.broadinstitute.sting.gatk.refdata.rodDbSNP;
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import org.broadinstitute.sting.gatk.refdata.rodGFF;
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import org.broadinstitute.sting.gatk.refdata.HapMapAlleleFrequenciesROD;
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import org.broadinstitute.sting.gatk.refdata.rodSAMPileup;
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import org.broadinstitute.sting.gatk.walkers.LocusWalker;
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import org.broadinstitute.sting.gatk.walkers.ReadWalker;
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import org.broadinstitute.sting.gatk.walkers.Walker;
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import org.broadinstitute.sting.gatk.traversals.*;
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import org.broadinstitute.sting.gatk.executive.MicroManager;
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import org.broadinstitute.sting.utils.FastaSequenceFile2;
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import org.broadinstitute.sting.utils.GenomeLoc;
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import org.broadinstitute.sting.utils.Utils;
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import org.broadinstitute.sting.utils.cmdLine.CommandLineProgram;
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import java.io.File;
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import java.io.PrintStream;
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import java.io.FileNotFoundException;
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import java.util.ArrayList;
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import java.util.List;
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public class GenomeAnalysisTK extends CommandLineProgram {
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public static GenomeAnalysisTK Instance = null;
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// parameters and their defaults
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public File INPUT_FILE = null;
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public String MAX_READS_ARG = "-1";
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public String STRICTNESS_ARG = "strict";
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public File REF_FILE_ARG = null;
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public String DEBUGGING_STR = null;
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public String REGION_STR = null;
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public String Analysis_Name = null;
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public String DBSNP_FILE = null;
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public String HAPMAP_FILE = null;
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public String HAPMAP_CHIP_FILE = null;
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public Boolean ENABLED_THREADED_IO = false;
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public Boolean UNSAFE = false;
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public String MAX_ON_FLY_SORTS = null;
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public String DOWNSAMPLE_FRACTION = null;
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public String DOWNSAMPLE_COVERAGE = null;
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public String INTERVALS_FILE = null;
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// added for mendelian walker.
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//TODO: when walkers can ask for their tracks this should be removed!
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public String MOTHER_GENOTYPE_FILE = null;
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public String FATHER_GENOTYPE_FILE = null;
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public String DAUGHTER_GENOTYPE_FILE = null;
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// our walker manager
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private WalkerManager walkerManager = null;
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public String pluginPathName = null;
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private TraversalEngine engine = null;
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public boolean DEBUGGING = false;
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public Boolean WALK_ALL_LOCI = false;
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public Boolean ENABLE_THREADING = false;
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/**
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* An output file presented to the walker.
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*/
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public String outFileName = null;
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/**
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* An error output file presented to the walker.
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*/
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public String errFileName = null;
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/**
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* A joint file for both 'normal' and error output presented to the walker.
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*/
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public String outErrFileName = null;
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/**
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* How many threads should be allocated to this analysis.
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*/
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public int numThreads = 1;
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/**
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* The output stream, initialized from OUTFILENAME / OUTERRFILENAME.
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* Used by the walker.
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*/
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public PrintStream out = System.out;
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/**
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* The output stream, initialized from ERRFILENAME / OUTERRFILENAME.
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* Used by the walker.
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*/
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public PrintStream err = System.err;
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/**
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* our log, which we want to capture anything from this class
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*/
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private static Logger logger = Logger.getLogger(GenomeAnalysisTK.class);
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public static ArrayList<String> ROD_BINDINGS = null;
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/**
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* setup our arguments, both required and optional
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* <p/>
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* Flags don't take an argument, the associated Boolean gets set to true if the flag appears on the command line.
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*/
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protected void setupArgs() {
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m_parser.addOptionalArg("input_file", "I", "SAM or BAM file", "INPUT_FILE");
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//m_parser.addRequiredArg("input_file", "I", "SAM or BAM file", "INPUT_FILE");
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m_parser.addOptionalArg("maximum_reads", "M", "Maximum number of reads to process before exiting", "MAX_READS_ARG");
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m_parser.addOptionalArg("validation_strictness", "S", "How strict should we be with validation (lenient|silent|strict)", "STRICTNESS_ARG");
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m_parser.addOptionalArg("reference_sequence", "R", "Reference sequence file", "REF_FILE_ARG");
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m_parser.addOptionalArg("genome_region", "L", "Genome region to operation on: from chr:start-end", "REGION_STR");
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m_parser.addRequiredArg("analysis_type", "T", "Type of analysis to run", "Analysis_Name");
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m_parser.addOptionalArg("DBSNP", "D", "DBSNP file", "DBSNP_FILE");
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m_parser.addOptionalArg("hapmap", "H", "Hapmap file", "HAPMAP_FILE");
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m_parser.addOptionalArg("hapmap_chip", "hc", "Hapmap chip file", "HAPMAP_CHIP_FILE");
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m_parser.addOptionalFlag("threaded_IO", "P", "If set, enables threaded I/O operations", "ENABLED_THREADED_IO");
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m_parser.addOptionalFlag("unsafe", "U", "If set, enables unsafe operations, nothing will be checked at runtime.", "UNSAFE");
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m_parser.addOptionalArg("sort_on_the_fly", "sort", "Maximum number of reads to sort on the fly", "MAX_ON_FLY_SORTS");
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m_parser.addOptionalArg("downsample_to_fraction", "dfrac", "Fraction [0.0-1.0] of reads to downsample to", "DOWNSAMPLE_FRACTION");
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m_parser.addOptionalArg("downsample_to_coverage", "dcov", "Coverage [integer] to downsample to", "DOWNSAMPLE_COVERAGE");
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m_parser.addOptionalArg("intervals_file", "V", "File containing list of genomic intervals to operate on. line := <contig> <start> <end>", "INTERVALS_FILE");
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m_parser.addOptionalFlag("all_loci", "A", "Should we process all loci, not just those covered by reads", "WALK_ALL_LOCI");
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m_parser.addOptionalArg("out", "o", "An output file presented to the walker. Will overwrite contents if file exists.", "outFileName" );
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m_parser.addOptionalArg("err", "e", "An error output file presented to the walker. Will overwrite contents if file exists.", "errFileName" );
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m_parser.addOptionalArg("outerr", "oe", "A joint file for 'normal' and error output presented to the walker. Will overwrite contents if file exists.", "outErrFileName");
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m_parser.addOptionalArg("numthreads", "nt", "How many threads should be allocated to running this analysis.", "numThreads");
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m_parser.addOptionalFlag("enablethreading", "et", "Enable experimental threading support.", "ENABLE_THREADING");
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//TODO: remove when walkers can ask for tracks
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m_parser.addOptionalArg("mother", "MOM", "Mother's genotype (SAM pileup)", "MOTHER_GENOTYPE_FILE");
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m_parser.addOptionalArg("father", "DAD", "Father's genotype (SAM pileup)", "FATHER_GENOTYPE_FILE");
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m_parser.addOptionalArg("daughter", "KID", "Daughter's genotype (SAM pileup)", "DAUGHTER_GENOTYPE_FILE");
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// --rodBind <name> <type> <file>
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Option rodBinder = OptionBuilder.withArgName("rodBind")
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.hasArgs()
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.withDescription( "Bind rod with <name> and <type> to <file>" )
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.create("B");
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m_parser.addOptionalArg(rodBinder, "ROD_BINDINGS");
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}
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/**
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* GATK can add arguments dynamically based on analysis type.
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* @return true
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*/
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@Override
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protected boolean canAddArgumentsDynamically() { return true; }
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/**
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* GATK provides the walker as an argument source. As a side-effect, initializes the walker variable.
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* @return List of walkers to load dynamically.
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*/
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@Override
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protected Class[] getArgumentSources() {
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if( Analysis_Name == null )
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throw new IllegalArgumentException("Must provide analysis name");
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walkerManager = new WalkerManager( pluginPathName );
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if( !walkerManager.doesWalkerExist(Analysis_Name) )
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throw new IllegalArgumentException("Invalid analysis name");
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return new Class[] { walkerManager.getWalkerClassByName(Analysis_Name) };
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}
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@Override
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protected String getArgumentSourceName( Class argumentSource ) {
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return WalkerManager.getWalkerName( (Class<Walker>)argumentSource );
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}
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/**
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* Required main method implementation.
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*/
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public static void main(String[] argv) {
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Instance = new GenomeAnalysisTK();
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start(Instance, argv);
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}
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protected int execute() {
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final boolean TEST_ROD = false;
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List<ReferenceOrderedData<? extends ReferenceOrderedDatum> > rods = new ArrayList<ReferenceOrderedData<? extends ReferenceOrderedDatum> >();
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if ( ROD_BINDINGS != null ) {
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System.out.printf("ROD BINDINGS are %s%n", Utils.join(":", ROD_BINDINGS));
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}
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if ( TEST_ROD ) {
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ReferenceOrderedData<rodGFF> gff = new ReferenceOrderedData<rodGFF>("test", new File("trunk/data/gFFTest.gff"), rodGFF.class );
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gff.testMe();
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//ReferenceOrderedData dbsnp = new ReferenceOrderedData(new File("trunk/data/dbSNP_head.txt"), rodDbSNP.class );
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ReferenceOrderedData<rodDbSNP> dbsnp = new ReferenceOrderedData<rodDbSNP>("dbSNP", new File("/Volumes/Users/mdepristo/broad/ATK/exampleSAMs/dbSNP_chr20.txt"), rodDbSNP.class );
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//dbsnp.testMe();
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rods.add(dbsnp); // { gff, dbsnp };
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} else {
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if ( DBSNP_FILE != null ) {
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ReferenceOrderedData<rodDbSNP> dbsnp = new ReferenceOrderedData<rodDbSNP>("dbSNP", new File(DBSNP_FILE), rodDbSNP.class );
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//dbsnp.testMe();
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rods.add(dbsnp); // { gff, dbsnp };
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}
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if ( HAPMAP_FILE != null ) {
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ReferenceOrderedData<HapMapAlleleFrequenciesROD> hapmap = new ReferenceOrderedData<HapMapAlleleFrequenciesROD>("hapmap", new File(HAPMAP_FILE), HapMapAlleleFrequenciesROD.class );
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//dbsnp.testMe();
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rods.add(hapmap); // { gff, dbsnp };
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}
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if ( HAPMAP_CHIP_FILE != null ) {
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ReferenceOrderedData<rodGFF> hapmapChip = new ReferenceOrderedData<rodGFF>("hapmap-chip", new File(HAPMAP_CHIP_FILE), rodGFF.class );
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rods.add(hapmapChip);
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}
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//TODO: remove when walkers can ask for tracks
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if ( MOTHER_GENOTYPE_FILE != null )
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rods.add( new ReferenceOrderedData<rodSAMPileup>("mother", new File(MOTHER_GENOTYPE_FILE), rodSAMPileup.class ) );
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if ( FATHER_GENOTYPE_FILE != null )
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rods.add( new ReferenceOrderedData<rodSAMPileup>("father", new File(FATHER_GENOTYPE_FILE), rodSAMPileup.class ) );
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if ( DAUGHTER_GENOTYPE_FILE != null )
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rods.add( new ReferenceOrderedData<rodSAMPileup>("daughter", new File(DAUGHTER_GENOTYPE_FILE), rodSAMPileup.class ) );
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}
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initializeOutputStreams();
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Walker<?,?> my_walker = null;
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try {
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my_walker = walkerManager.createWalkerByName( Analysis_Name );
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}
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catch( InstantiationException ex ) {
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throw new RuntimeException( "Unable to instantiate walker.", ex );
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}
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catch( IllegalAccessException ex ) {
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throw new RuntimeException( "Unable to access walker", ex );
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}
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MicroManager microManager = null;
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// Try to get the walker specified
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try {
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LocusWalker<?, ?> walker = (LocusWalker<?, ?>) my_walker;
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if ( INPUT_FILE == null ) {
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if ( walker.requiresReads() )
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Utils.scareUser(String.format("Analysis %s requires reads, but none were given", Analysis_Name));
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this.engine = new TraverseByReference(null, REF_FILE_ARG, rods);
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} else {
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if ( walker.cannotHandleReads() )
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Utils.scareUser(String.format("Analysis %s doesn't support SAM/BAM reads, but a read file %s was provided", Analysis_Name, INPUT_FILE));
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if ( WALK_ALL_LOCI ) {
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// TODO: Temporary debugging code. Activate the new debugging code only when the MicroManager
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// is not filtered.
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if( ENABLE_THREADING && REGION_STR == null ) {
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logger.warn("Preliminary threading support enabled");
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microManager = new MicroManager( INPUT_FILE, REF_FILE_ARG, numThreads );
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this.engine = microManager.getTraversalEngine();
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}
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else {
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this.engine = new TraverseByLociByReference(INPUT_FILE, REF_FILE_ARG, rods);
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}
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}
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else
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this.engine = new TraverseByLoci(INPUT_FILE, REF_FILE_ARG, rods);
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}
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}
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catch (java.lang.ClassCastException e) {
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// I guess we're a read walker LOL
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ReadWalker<?, ?> walker = (ReadWalker<?, ?>) my_walker;
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this.engine = new TraverseByReads(INPUT_FILE, REF_FILE_ARG, rods);
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}
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// Prepare the sort ordering w.r.t. the sequence dictionary
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if (REF_FILE_ARG != null) {
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final ReferenceSequenceFile refFile = ReferenceSequenceFileFactory.getReferenceSequenceFile(REF_FILE_ARG);
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GenomeLoc.setupRefContigOrdering(refFile);
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}
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// Determine the validation stringency. Default to ValidationStringency.STRICT.
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ValidationStringency strictness;
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try {
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strictness = Enum.valueOf(ValidationStringency.class, STRICTNESS_ARG);
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}
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catch( IllegalArgumentException ex ) {
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strictness = ValidationStringency.STRICT;
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}
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logger.info("Strictness is " + strictness);
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engine.setStrictness(strictness);
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engine.setDebugging(!(DEBUGGING_STR == null || DEBUGGING_STR.toLowerCase().equals("true")));
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engine.setMaxReads(Integer.parseInt(MAX_READS_ARG));
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if (REGION_STR != null) {
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engine.setLocation(REGION_STR);
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}
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if (INTERVALS_FILE != null) {
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engine.setLocationFromFile(INTERVALS_FILE);
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}
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if (MAX_ON_FLY_SORTS != null) {
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engine.setSortOnFly(Integer.parseInt(MAX_ON_FLY_SORTS));
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}
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if (DOWNSAMPLE_FRACTION != null) {
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engine.setDownsampleByFraction(Double.parseDouble(DOWNSAMPLE_FRACTION));
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}
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if (DOWNSAMPLE_COVERAGE != null) {
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engine.setDownsampleByCoverage(Integer.parseInt(DOWNSAMPLE_COVERAGE));
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}
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engine.setSafetyChecking(!UNSAFE);
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engine.setThreadedIO(ENABLED_THREADED_IO);
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engine.setWalkOverAllSites(WALK_ALL_LOCI);
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engine.initialize();
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if( microManager != null ) {
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List<GenomeLoc> locations = GenomeLoc.parseGenomeLocs( REGION_STR );
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microManager.execute( my_walker, locations );
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}
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else
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engine.traverse(my_walker);
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return 0;
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}
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/**
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* Initialize the output streams as specified by the user.
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*/
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private void initializeOutputStreams() {
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if( outErrFileName != null && (outFileName != null || errFileName != null) )
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throw new IllegalArgumentException("Can't set output/error output file with either out file name or err file name");
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try {
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if( outErrFileName != null ) {
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PrintStream outErrStream = new PrintStream( outErrFileName );
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out = outErrStream;
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err = outErrStream;
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}
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if ( outFileName != null ) {
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out = new PrintStream( outFileName );
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}
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if( errFileName != null ) {
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err = new PrintStream( errFileName );
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}
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}
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catch( FileNotFoundException ex ) {
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throw new RuntimeException("Unable to open a walker output file.", ex);
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}
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}
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/**
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* An inappropriately placed validation and performance testing routine for jumping
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* around in the fasta sequence file.
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* @param refFileName
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*/
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private static void testNewReferenceFeatures(final File refFileName) {
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final FastaSequenceFile2 refFile = new FastaSequenceFile2(refFileName);
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GenomeLoc.setupRefContigOrdering(refFile);
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List<SAMSequenceRecord> refContigs = refFile.getSequenceDictionary().getSequences();
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/*
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for ( SAMSequenceRecord refContig: refContigs ) {
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System.out.printf(" Traversing from chr1 to %s would require jumping %d bytes%n",
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refContig.getSequenceName(), refFile.getDistanceBetweenContigs("chr1", refContig.getSequenceName()));
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}
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*/
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String lastContig = null;
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List<Double> timings = new ArrayList<Double>();
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for ( SAMSequenceRecord startContig : refFile.getSequenceDictionary().getSequences() ) {
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final String startContigName = startContig.getSequenceName();
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for ( SAMSequenceRecord targetContig : refFile.getSequenceDictionary().getSequences() ) {
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refFile.seekToContig(startContigName, true);
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logger.info(String.format("Seeking: current=%s, target=%s%n", startContigName, targetContig.getSequenceName()));
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long lastTime = System.currentTimeMillis();
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final boolean success = refFile.seekToContig(targetContig.getSequenceName(), true);
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long curTime = System.currentTimeMillis();
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final double elapsed = (curTime - lastTime) / 1000.0;
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timings.add(elapsed);
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logger.info(String.format(" -> Elapsed time %.2f, averaging %.2f sec / seek for %d seeks%n",
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elapsed, Utils.averageDouble(timings), timings.size()));
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if ( ! success ) {
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logger.error(String.format("Failured to seek to %s from %s%n", targetContig.getSequenceName(), lastContig ));
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}
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//System.exit(1);
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}
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}
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System.exit(1);
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// code for randomly sampling the seeks
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// Random rnd = new Random();
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// String lastContig = null;
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// List<Double> timings = new ArrayList<Double>();
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// final int N_SAMPLES = 1000;
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// //try { refFile.seekToContig("chr3"); } catch ( IOException e ) {}
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// for ( int i = 0; i < N_SAMPLES; i++ ) {
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// final int nextIndex = rnd.nextInt(refContigs.size());
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// String nextContig = refFile.getSequenceDictionary().getSequence(nextIndex).getSequenceName();
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// //nextContig = "chr2";
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// try {
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// System.out.printf("Seeking: current=%s, target=%s%n", refFile.getContigName(), nextContig);
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// long lastTime = System.currentTimeMillis();
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// final boolean success = refFile.seekToContig(nextContig, true);
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// long curTime = System.currentTimeMillis();
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// final double elapsed = (curTime - lastTime) / 1000.0;
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// timings.add(elapsed);
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// System.out.printf(" -> Elapsed time %.2f, averaging %.2f sec / seek for %d seeks%n",
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// elapsed, Utils.averageDouble(timings), timings.size());
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//
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// if ( ! success ) {
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// System.out.printf("Failured to seek to %s from %s%n", nextContig, lastContig );
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// }
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// //System.exit(1);
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// } catch ( IOException e ) {
|
|
// System.out.printf("Failured to seek to %s from %s%n", nextContig, lastContig );
|
|
// e.printStackTrace();
|
|
// }
|
|
//
|
|
// lastContig = nextContig;
|
|
// }
|
|
// System.exit(1);
|
|
|
|
/*
|
|
final String targetChr = "chr10";
|
|
try {
|
|
refFile.seekToContig(targetChr);
|
|
} catch ( IOException e ){
|
|
System.out.printf("Failured to seek to %s%n", targetChr);
|
|
e.printStackTrace();
|
|
}
|
|
System.exit(1);
|
|
*/
|
|
|
|
//List<Double> timings = new ArrayList<Double>();
|
|
final long startTime = System.currentTimeMillis();
|
|
long lastTime = System.currentTimeMillis();
|
|
|
|
int i = 0;
|
|
String prevNextContigName = null;
|
|
logger.info(String.format("Walking reference sequence:%n"));
|
|
for ( SAMSequenceRecord refContig: refContigs ) {
|
|
long curTime = System.currentTimeMillis();
|
|
ReferenceSequence contig = refFile.nextSequence();
|
|
final double elapsed = (curTime - lastTime) / 1000.0;
|
|
timings.add(elapsed);
|
|
logger.info(String.format("%2d : expected %s contig, found %s with next of %s after %.2f seconds, average is %.2f%n", i,
|
|
refContig.getSequenceName(), contig.getName(), refFile.getNextContigName(), elapsed, Utils.averageDouble(timings)));
|
|
if ( prevNextContigName != null && contig.getName() != null && ! prevNextContigName.equals(contig.getName()) )
|
|
throw new RuntimeIOException(String.format("Unexpected contig ordering %s was expected next, but I found %s?",
|
|
prevNextContigName, contig.getName()));
|
|
|
|
prevNextContigName = refFile.getNextContigName();
|
|
lastTime = curTime;
|
|
i++;
|
|
|
|
logger.info(String.format(" Traversing from chr1 to %s would require jumping %d bytes%n",
|
|
contig.getName(), refFile.getDistanceBetweenContigs("chr1", contig.getName())));
|
|
}
|
|
}
|
|
|
|
public SAMFileReader getSamReader() { return this.engine.getSamReader(); }
|
|
}
|