/**
* Instantiates multiple underlying SAM writes, one per input SAM reader registered with GATK
* engine (those will be retrieved from <code>toolkit</code>). The output file names will be
* generated automatically by stripping ".sam" or ".bam" off the input file name and adding ext
* instead (e.g. ".cleaned.bam"). onto a unique output file name.
*
* @param toolkit
* @param ext
*/
public void setupByReader(
GenomeAnalysisEngine toolkit,
String ext,
SAMFileHeader.SortOrder order,
boolean presorted,
boolean indexOnTheFly,
boolean generateMD5,
SAMProgramRecord pRecord) {
for (SAMReaderID rid : toolkit.getReadsDataSource().getReaderIDs()) {
String fName = toolkit.getReadsDataSource().getSAMFile(rid).getName();
String outName;
int pos;
if (fName.toUpperCase().endsWith(".BAM")) pos = fName.toUpperCase().lastIndexOf(".BAM");
else {
if (fName.toUpperCase().endsWith(".SAM")) pos = fName.toUpperCase().lastIndexOf(".SAM");
else
throw new UserException.BadInput(
"Input file name " + fName + " does not end with .sam or .bam");
}
String prefix = fName.substring(0, pos);
outName = prefix + ext;
if (writerMap.containsKey(rid))
throw new StingException(
"nWayOut mode: Reader id for input sam file " + fName + " is already registered");
addWriter(rid, outName, order, presorted, indexOnTheFly, generateMD5, pRecord);
}
}
@Requires({
"toolkit != null",
"UAC != null",
"logger != null",
"samples != null && samples.size() > 0",
"ploidy>0"
})
public UnifiedGenotyperEngine(
GenomeAnalysisEngine toolkit,
UnifiedArgumentCollection UAC,
Logger logger,
PrintStream verboseWriter,
VariantAnnotatorEngine engine,
Set<String> samples,
int ploidy) {
this.BAQEnabledOnCMDLine = toolkit.getArguments().BAQMode != BAQ.CalculationMode.OFF;
genomeLocParser = toolkit.getGenomeLocParser();
this.samples = new TreeSet<String>(samples);
// note that, because we cap the base quality by the mapping quality, minMQ cannot be less than
// minBQ
this.UAC = UAC;
this.logger = logger;
this.verboseWriter = verboseWriter;
this.annotationEngine = engine;
this.ploidy = ploidy;
this.N = samples.size() * ploidy;
log10AlleleFrequencyPriorsSNPs = new double[N + 1];
log10AlleleFrequencyPriorsIndels = new double[N + 1];
computeAlleleFrequencyPriors(N, log10AlleleFrequencyPriorsSNPs, UAC.heterozygosity);
computeAlleleFrequencyPriors(N, log10AlleleFrequencyPriorsIndels, UAC.INDEL_HETEROZYGOSITY);
filter.add(LOW_QUAL_FILTER_NAME);
}
/**
* Instantiates multiple underlying SAM writes, one per input SAM reader registered with GATK
* engine (those will be retrieved from <code>toolkit</code>). The <code>in2out</code> map must
* contain an entry for each input filename and map it onto a unique output file name.
*
* @param toolkit
* @param in2out
*/
public void setupByReader(
GenomeAnalysisEngine toolkit,
Map<String, String> in2out,
SAMFileHeader.SortOrder order,
boolean presorted,
boolean indexOnTheFly,
boolean generateMD5,
SAMProgramRecord pRecord) {
if (in2out == null)
throw new StingException("input-output bam filename map for n-way-out writing is NULL");
for (SAMReaderID rid : toolkit.getReadsDataSource().getReaderIDs()) {
String fName = toolkit.getReadsDataSource().getSAMFile(rid).getName();
String outName;
if (!in2out.containsKey(fName))
throw new UserException.BadInput(
"Input-output bam filename map does not contain an entry for the input file " + fName);
outName = in2out.get(fName);
if (writerMap.containsKey(rid))
throw new StingException(
"nWayOut mode: Reader id for input sam file "
+ fName
+ " is already registered; "
+ "map file likely contains multiple entries for this input file");
addWriter(rid, outName, order, presorted, indexOnTheFly, generateMD5, pRecord);
}
}
public void addAlignment(SAMRecord samRecord) {
final SAMReaderID id = toolkit.getReaderIDForRead(samRecord);
String rg = samRecord.getStringAttribute("RG");
if (rg != null) {
String rg_orig = toolkit.getReadsDataSource().getOriginalReadGroupId(rg);
samRecord.setAttribute("RG", rg_orig);
}
addAlignment(samRecord, id);
}
/**
* Create random read qualities
*
* @param length the length of the read
* @param allowNs whether or not to allow N's in the read
* @return an array with randomized bases (A-N) with equal probability
*/
public static byte[] createRandomReadBases(int length, boolean allowNs) {
Random random = GenomeAnalysisEngine.getRandomGenerator();
int numberOfBases = allowNs ? 5 : 4;
byte[] bases = new byte[length];
for (int i = 0; i < length; i++) {
switch (random.nextInt(numberOfBases)) {
case 0:
bases[i] = 'A';
break;
case 1:
bases[i] = 'C';
break;
case 2:
bases[i] = 'G';
break;
case 3:
bases[i] = 'T';
break;
case 4:
bases[i] = 'N';
break;
default:
throw new ReviewedStingException("Something went wrong, this is just impossible");
}
}
return bases;
}
/**
* Gets the appropriately formatted header for a VCF file describing this GATK run
*
* @param engine the GATK engine that holds the walker name, GATK version, and other information
* @param argumentSources contains information on the argument values provided to the GATK for
* converting to a command line string. Should be provided from the data in the parsing
* engine. Can be empty in which case the command line will be the empty string.
* @return VCF header line describing this run of the GATK.
*/
public static VCFHeaderLine getCommandLineArgumentHeaderLine(
final GenomeAnalysisEngine engine, final Collection<Object> argumentSources) {
if (engine == null) throw new IllegalArgumentException("engine cannot be null");
if (argumentSources == null)
throw new IllegalArgumentException("argumentSources cannot be null");
final Map<String, String> attributes = new LinkedHashMap<>();
attributes.put("ID", engine.getWalkerName());
attributes.put("Version", CommandLineGATK.getVersionNumber());
final Date date = new Date();
attributes.put("Date", date.toString());
attributes.put("Epoch", Long.toString(date.getTime()));
attributes.put(
"CommandLineOptions",
engine.createApproximateCommandLineArgumentString(argumentSources.toArray()));
return new VCFSimpleHeaderLine(GATK_COMMAND_LINE_KEY, attributes);
}
// ---------------------------------------------------------------------------------------------------------
//
// Public interface functions
//
// ---------------------------------------------------------------------------------------------------------
@Requires({"toolkit != null", "UAC != null"})
public UnifiedGenotyperEngine(GenomeAnalysisEngine toolkit, UnifiedArgumentCollection UAC) {
this(
toolkit,
UAC,
Logger.getLogger(UnifiedGenotyperEngine.class),
null,
null,
SampleUtils.getSAMFileSamples(toolkit.getSAMFileHeader()),
VariantContextUtils.DEFAULT_PLOIDY);
}
private void randomlyAddVariant(int rank, VariantContext vc, byte refBase) {
if (nVariantsAdded < numRandom) variantArray[nVariantsAdded++] = new RandomVariantStructure(vc);
else {
double v = GenomeAnalysisEngine.getRandomGenerator().nextDouble();
double t = (1.0 / (rank - numRandom + 1));
if (v < t) {
variantArray[positionToAdd].set(vc);
nVariantsAdded++;
positionToAdd = nextCircularPosition(positionToAdd);
}
}
}
/**
* Creates a program record (@PG) tag
*
* @param toolkit the engine
* @param walker the walker object (so we can extract the command line)
* @param PROGRAM_RECORD_NAME the name for the PG tag
* @return a program record for the tool
*/
public static SAMProgramRecord createProgramRecord(
GenomeAnalysisEngine toolkit, Object walker, String PROGRAM_RECORD_NAME) {
final SAMProgramRecord programRecord = new SAMProgramRecord(PROGRAM_RECORD_NAME);
final ResourceBundle headerInfo = TextFormattingUtils.loadResourceBundle("StingText");
try {
final String version = headerInfo.getString("org.broadinstitute.sting.gatk.version");
programRecord.setProgramVersion(version);
} catch (MissingResourceException e) {
// couldn't care less if the resource is missing...
}
programRecord.setCommandLine(
toolkit.createApproximateCommandLineArgumentString(toolkit, walker));
return programRecord;
}
public static Map<String, VCFHeader> getVCFHeadersFromRodPrefix(
GenomeAnalysisEngine toolkit, String prefix) {
Map<String, VCFHeader> data = new HashMap<String, VCFHeader>();
// iterate to get all of the sample names
List<ReferenceOrderedDataSource> dataSources = toolkit.getRodDataSources();
for (ReferenceOrderedDataSource source : dataSources) {
// ignore the rod if lacks the prefix
if (!source.getName().startsWith(prefix)) continue;
if (source.getHeader() != null && source.getHeader() instanceof VCFHeader)
data.put(source.getName(), (VCFHeader) source.getHeader());
}
return data;
}
public static Map<String, VCFHeader> getVCFHeadersFromRods(
GenomeAnalysisEngine toolkit, Collection<String> rodNames) {
Map<String, VCFHeader> data = new HashMap<String, VCFHeader>();
// iterate to get all of the sample names
List<ReferenceOrderedDataSource> dataSources = toolkit.getRodDataSources();
for (ReferenceOrderedDataSource source : dataSources) {
// ignore the rod if it's not in our list
if (rodNames != null && !rodNames.contains(source.getName())) continue;
if (source.getHeader() != null && source.getHeader() instanceof VCFHeader)
data.put(source.getName(), (VCFHeader) source.getHeader());
}
return data;
}
@Test(enabled = true)
public void testGetBasesReverseComplement() {
int iterations = 1000;
Random random = GenomeAnalysisEngine.getRandomGenerator();
while (iterations-- > 0) {
final int l = random.nextInt(1000);
GATKSAMRecord read = GATKSAMRecord.createRandomRead(l);
byte[] original = read.getReadBases();
byte[] reconverted = new byte[l];
String revComp = ReadUtils.getBasesReverseComplement(read);
for (int i = 0; i < l; i++) {
reconverted[l - 1 - i] = BaseUtils.getComplement((byte) revComp.charAt(i));
}
Assert.assertEquals(reconverted, original);
}
}
private void addWriter(
SAMReaderID id,
String outName,
SAMFileHeader.SortOrder order,
boolean presorted,
boolean indexOnTheFly,
boolean generateMD5,
SAMProgramRecord programRecord) {
File f = new File(outName);
SAMFileHeader header =
Utils.setupWriter(
toolkit, toolkit.getSAMFileHeader(id), KEEP_ALL_PG_RECORDS, programRecord);
SAMFileWriterFactory factory = new SAMFileWriterFactory();
factory.setCreateIndex(indexOnTheFly);
factory.setCreateMd5File(generateMD5);
SAMFileWriter sw = factory.makeSAMOrBAMWriter(header, presorted, f);
writerMap.put(id, sw);
}
private void testPerSampleEqualsFlat(
final String bam1,
final String bam2,
final String persampleFile,
final Double downsampling,
final String md5) {
final String command =
baseCommand3
+ " -I "
+ ArtificalBAMLocation
+ bam1
+ " -I "
+ ArtificalBAMLocation
+ bam2
+ " -o %s ";
WalkerTestSpec spec =
new WalkerTestSpec(command + " -contaminationFile " + persampleFile, 1, Arrays.asList(md5));
final Random rnd = GenomeAnalysisEngine.getRandomGenerator();
rnd.setSeed(123451); // so that the two test cases have a hope of giving the same result
executeTest(
"test contamination on Artificial Contamination, with per-sample file on "
+ bam1
+ " and "
+ bam2
+ " with "
+ persampleFile,
spec);
spec =
new WalkerTestSpec(
command + "-contamination " + downsampling.toString(), 1, Arrays.asList(md5));
rnd.setSeed(123451); // so that the two test cases have a hope of giving the same result
executeTest(
"test contamination on Artificial Contamination, with flat contamination on "
+ bam1
+ " and "
+ bam2
+ " with "
+ downsampling.toString(),
spec);
}
/**
* Gets the header fields from all VCF rods input by the user
*
* @param toolkit GATK engine
* @param rodNames names of rods to use, or null if we should use all possible ones
* @return a set of all fields
*/
public static Set<VCFHeaderLine> getHeaderFields(
GenomeAnalysisEngine toolkit, Collection<String> rodNames) {
// keep a map of sample name to occurrences encountered
TreeSet<VCFHeaderLine> fields = new TreeSet<VCFHeaderLine>();
// iterate to get all of the sample names
List<ReferenceOrderedDataSource> dataSources = toolkit.getRodDataSources();
for (ReferenceOrderedDataSource source : dataSources) {
// ignore the rod if it's not in our list
if (rodNames != null && !rodNames.contains(source.getName())) continue;
if (source.getRecordType().equals(VariantContext.class)) {
VCFHeader header = (VCFHeader) source.getHeader();
if (header != null) fields.addAll(header.getMetaDataInSortedOrder());
}
}
return fields;
}
/**
* Create random read qualities
*
* @param length the length of the read
* @return an array with randomized base qualities between 0 and 50
*/
public static byte[] createRandomReadQuals(int length) {
Random random = GenomeAnalysisEngine.getRandomGenerator();
byte[] quals = new byte[length];
for (int i = 0; i < length; i++) quals[i] = (byte) random.nextInt(50);
return quals;
}
public SAMFileHeader getFileHeader() {
return toolkit.getSAMFileHeader();
}
/**
* Add / replace the contig header lines in the VCFHeader with the information in the GATK engine
*
* @param header the header to update
* @param engine the GATK engine containing command line arguments and the master sequence
* dictionary
*/
public static VCFHeader withUpdatedContigs(
final VCFHeader header, final GenomeAnalysisEngine engine) {
return VCFUtils.withUpdatedContigs(
header, engine.getArguments().referenceFile, engine.getMasterSequenceDictionary());
}
/**
* Subset VC record if necessary and emit the modified record (provided it satisfies criteria for
* printing)
*
* @param tracker the ROD tracker
* @param ref reference information
* @param context alignment info
* @return 1 if the record was printed to the output file, 0 if otherwise
*/
@Override
public Integer map(RefMetaDataTracker tracker, ReferenceContext ref, AlignmentContext context) {
if (tracker == null) return 0;
Collection<VariantContext> vcs =
tracker.getValues(variantCollection.variants, context.getLocation());
if (vcs == null || vcs.size() == 0) {
return 0;
}
for (VariantContext vc : vcs) {
if (MENDELIAN_VIOLATIONS) {
boolean foundMV = false;
for (MendelianViolation mv : mvSet) {
if (mv.isViolation(vc)) {
foundMV = true;
// System.out.println(vc.toString());
if (outMVFile != null)
outMVFileStream.format(
"MV@%s:%d. REF=%s, ALT=%s, AC=%d, momID=%s, dadID=%s, childID=%s, momG=%s, momGL=%s, dadG=%s, dadGL=%s, "
+ "childG=%s childGL=%s\n",
vc.getChr(),
vc.getStart(),
vc.getReference().getDisplayString(),
vc.getAlternateAllele(0).getDisplayString(),
vc.getChromosomeCount(vc.getAlternateAllele(0)),
mv.getSampleMom(),
mv.getSampleDad(),
mv.getSampleChild(),
vc.getGenotype(mv.getSampleMom()).toBriefString(),
vc.getGenotype(mv.getSampleMom()).getLikelihoods().getAsString(),
vc.getGenotype(mv.getSampleDad()).toBriefString(),
vc.getGenotype(mv.getSampleMom()).getLikelihoods().getAsString(),
vc.getGenotype(mv.getSampleChild()).toBriefString(),
vc.getGenotype(mv.getSampleChild()).getLikelihoods().getAsString());
}
}
if (!foundMV) break;
}
if (DISCORDANCE_ONLY) {
Collection<VariantContext> compVCs =
tracker.getValues(discordanceTrack, context.getLocation());
if (!isDiscordant(vc, compVCs)) return 0;
}
if (CONCORDANCE_ONLY) {
Collection<VariantContext> compVCs =
tracker.getValues(concordanceTrack, context.getLocation());
if (!isConcordant(vc, compVCs)) return 0;
}
if (alleleRestriction.equals(NumberAlleleRestriction.BIALLELIC) && !vc.isBiallelic())
continue;
if (alleleRestriction.equals(NumberAlleleRestriction.MULTIALLELIC) && vc.isBiallelic())
continue;
if (!selectedTypes.contains(vc.getType())) continue;
VariantContext sub = subsetRecord(vc, samples);
if ((sub.isPolymorphic() || !EXCLUDE_NON_VARIANTS)
&& (!sub.isFiltered() || !EXCLUDE_FILTERED)) {
for (VariantContextUtils.JexlVCMatchExp jexl : jexls) {
if (!VariantContextUtils.match(sub, jexl)) {
return 0;
}
}
if (SELECT_RANDOM_NUMBER) {
randomlyAddVariant(++variantNumber, sub, ref.getBase());
} else if (!SELECT_RANDOM_FRACTION
|| (GenomeAnalysisEngine.getRandomGenerator().nextDouble() < fractionRandom)) {
vcfWriter.add(sub);
}
}
}
return 1;
}
public void writeBeagleOutput(
VariantContext preferredVC, VariantContext otherVC, boolean isValidationSite, double prior) {
GenomeLoc currentLoc =
VariantContextUtils.getLocation(getToolkit().getGenomeLocParser(), preferredVC);
StringBuffer beagleOut = new StringBuffer();
String marker = String.format("%s:%d ", currentLoc.getContig(), currentLoc.getStart());
beagleOut.append(marker);
if (markers != null)
markers.append(marker).append("\t").append(Integer.toString(markerCounter++)).append("\t");
for (Allele allele : preferredVC.getAlleles()) {
String bglPrintString;
if (allele.isNoCall() || allele.isNull()) bglPrintString = "-";
else bglPrintString = allele.getBaseString(); // get rid of * in case of reference allele
beagleOut.append(String.format("%s ", bglPrintString));
if (markers != null) markers.append(bglPrintString).append("\t");
}
if (markers != null) markers.append("\n");
GenotypesContext preferredGenotypes = preferredVC.getGenotypes();
GenotypesContext otherGenotypes = goodSite(otherVC) ? otherVC.getGenotypes() : null;
for (String sample : samples) {
boolean isMaleOnChrX = CHECK_IS_MALE_ON_CHR_X && getSample(sample).getGender() == Gender.MALE;
Genotype genotype;
boolean isValidation;
// use sample as key into genotypes structure
if (preferredGenotypes.containsSample(sample)) {
genotype = preferredGenotypes.get(sample);
isValidation = isValidationSite;
} else if (otherGenotypes != null && otherGenotypes.containsSample(sample)) {
genotype = otherGenotypes.get(sample);
isValidation = !isValidationSite;
} else {
// there is magically no genotype for this sample.
throw new StingException(
"Sample "
+ sample
+ " arose with no genotype in variant or validation VCF. This should never happen.");
}
/*
* Use likelihoods if: is validation, prior is negative; or: is not validation, has genotype key
*/
double[] log10Likelihoods = null;
if ((isValidation && prior < 0.0) || genotype.hasLikelihoods()) {
log10Likelihoods = genotype.getLikelihoods().getAsVector();
// see if we need to randomly mask out genotype in this position.
if (GenomeAnalysisEngine.getRandomGenerator().nextDouble() <= insertedNoCallRate) {
// we are masking out this genotype
log10Likelihoods =
isMaleOnChrX ? HAPLOID_FLAT_LOG10_LIKELIHOODS : DIPLOID_FLAT_LOG10_LIKELIHOODS;
}
if (isMaleOnChrX) {
log10Likelihoods[1] = -255; // todo -- warning this is dangerous for multi-allele case
}
}
/** otherwise, use the prior uniformly */
else if (!isValidation && genotype.isCalled() && !genotype.hasLikelihoods()) {
// hack to deal with input VCFs with no genotype likelihoods. Just assume the called
// genotype
// is confident. This is useful for Hapmap and 1KG release VCFs.
double AA = (1.0 - prior) / 2.0;
double AB = (1.0 - prior) / 2.0;
double BB = (1.0 - prior) / 2.0;
if (genotype.isHomRef()) {
AA = prior;
} else if (genotype.isHet()) {
AB = prior;
} else if (genotype.isHomVar()) {
BB = prior;
}
log10Likelihoods = MathUtils.toLog10(new double[] {AA, isMaleOnChrX ? 0.0 : AB, BB});
} else {
log10Likelihoods =
isMaleOnChrX ? HAPLOID_FLAT_LOG10_LIKELIHOODS : DIPLOID_FLAT_LOG10_LIKELIHOODS;
}
writeSampleLikelihoods(beagleOut, preferredVC, log10Likelihoods);
}
beagleWriter.println(beagleOut.toString());
}
