Java Code Examples for htsjdk.samtools.SAMRecord#setReadString()

@Test(dataProvider="clipPairedTestData")
public void testPairedEndClip(final String testName, final String read1, final String read2, final AdapterPair expected) {

    final SAMRecord rec1 = new SAMRecord(new SAMFileHeader());
    rec1.setReadString(read1);
    rec1.setFirstOfPairFlag(true);
    final SAMRecord rec2 = new SAMRecord(new SAMFileHeader());
    rec2.setReadString(read2);
    rec2.setSecondOfPairFlag(true);

    final AdapterPair result = ClippingUtility.adapterTrimIlluminaPairedReads(rec1, rec2,
            IlluminaAdapterPair.INDEXED, IlluminaAdapterPair.PAIRED_END);
    if (result != null) {
        Assert.assertEquals(result.getName(), expected.getName(), testName);
    }
    else {
        Assert.assertNull(expected, testName);
    }
}
 

private SAMRecord createSamRecord(final SAMFileHeader header, final String baseName, final FastqRecord frec, final boolean paired) {
    final SAMRecord srec = new SAMRecord(header);
    srec.setReadName(baseName);
    srec.setReadString(frec.getReadString());
    srec.setReadUnmappedFlag(true);
    srec.setAttribute(ReservedTagConstants.READ_GROUP_ID, READ_GROUP_NAME);
    final byte[] quals = StringUtil.stringToBytes(frec.getBaseQualityString());
    convertQuality(quals, QUALITY_FORMAT);
    for (final byte qual : quals) {
        final int uQual = qual & 0xff;
        if (uQual < MIN_Q || uQual > MAX_Q) {
            throw new PicardException("Base quality " + uQual + " is not in the range " + MIN_Q + ".." +
            MAX_Q + " for read " + frec.getReadHeader());
        }
    }
    srec.setBaseQualities(quals);

    if (paired) {
        srec.setReadPairedFlag(true);
        srec.setMateUnmappedFlag(true);
    }
    return srec ;
}
 

private static SAMRecord createRecord(int position, String ref) {
  final SAMFileHeader header = new SAMFileHeader();
  header.getSequenceDictionary().addSequence(new SAMSequenceRecord("A", 100000));
  header.getSequenceDictionary().addSequence(new SAMSequenceRecord("B", 100000));
  final SAMRecord rec = new SAMRecord(header);
  rec.setReferenceName(ref);
  rec.setReadName("read");
  rec.setReadString("ACGT");
  rec.setBaseQualityString("####");
  if (ref.equals(SAMRecord.NO_ALIGNMENT_REFERENCE_NAME)) {
    rec.setReadUnmappedFlag(true);
  } else {
    rec.setCigarString("4=");
    rec.setAlignmentStart(position);
  }
  return rec;
}
 

private SAMRecord createSamRecordWithAdapterSequence(final int readLength, final IlluminaAdapterPair adapterPair, final int adapterPosition) {
    final String adapterString = adapterPair.get3PrimeAdapterInReadOrder();
    final int replacementLength = Math.min(adapterString.length(), readLength - adapterPosition);
    final String adapterSubstring = adapterString.substring(0, replacementLength);
    final String readBases = replaceSubstring(makeBogusReadString(readLength), adapterSubstring, adapterPosition, adapterSubstring.length());
    final SAMRecord rec = new SAMRecord(null);
    rec.setReadString(readBases);
    return rec;
}
 

@NotNull
public static SAMRecord buildSamRecord(final int alignmentStart, @NotNull final String cigar, @NotNull final String readString,
        @NotNull final String qualities, final boolean negativeStrand) {
    final SAMRecord record = new SAMRecord(null);
    record.setAlignmentStart(alignmentStart);
    record.setCigarString(cigar);
    record.setReadString(readString);
    record.setReadNegativeStrandFlag(negativeStrand);
    record.setBaseQualityString(qualities);
    record.setMappingQuality(20);
    record.setDuplicateReadFlag(false);
    record.setReadUnmappedFlag(false);
    return record;
}
 

public void addAlignment(int sampleIdx, SAMRecordWrapper samRecord, int chromosomeChunkIdx) {
	Feature chunk = this.chromosomeChunker.getChunks().get(chromosomeChunkIdx);
	
	SAMRecord read = samRecord.getSamRecord();
	
	// Only output reads with original start pos within specified chromosomeChunk
	// Avoids reads being written in 2 different chunks
	
	int origAlignmentStart = read.getAlignmentStart();
	String yo = read.getStringAttribute("YO");
	if (yo != null) {
		String[] fields = yo.split(":");
		origAlignmentStart = Integer.parseInt(fields[1]);
	}
	
	if (origAlignmentStart >= chunk.getStart() && origAlignmentStart <= chunk.getEnd()) {
		
		if (samRecord.isUnalignedRc() && read.getReadUnmappedFlag()) {
			// This read was reverse complemented, but not updated.
			// Change it back to its original state.
			read.setReadString(rc.reverseComplement(read.getReadString()));
			read.setBaseQualityString(rc.reverse(read.getBaseQualityString()));
			read.setReadNegativeStrandFlag(!read.getReadNegativeStrandFlag());
		}
		
		writers[sampleIdx][chromosomeChunkIdx].addAlignment(read);
	}
}
 

/**
 * Replace hard clips with soft clips.
 */
public static void replaceHardClips(SAMRecord read) {
	Cigar cigar = read.getCigar();
	
	if (cigar.getCigarElements().size() > 0) {
		CigarElement firstElement = cigar.getCigarElement(0);
		CigarElement lastElement  = cigar.getCigarElement(cigar.numCigarElements()-1);
		
		if ((firstElement.getOperator() == CigarOperator.H) ||
			(lastElement.getOperator() == CigarOperator.H)) {
			
			Cigar newCigar = new Cigar();
			
			boolean isFirst = true;
			
			for (CigarElement element : cigar.getCigarElements()) {
				if (element.getOperator() != CigarOperator.H) {
					newCigar.add(element);
				} else {
					CigarElement softClip = new CigarElement(element.getLength(), CigarOperator.S);
					newCigar.add(softClip);
					
					if (isFirst) {
						read.setReadString(padBases(element.getLength()) + read.getReadString());
					} else {
						read.setReadString(read.getReadString() + padBases(element.getLength()));							
					}
				}
				
				isFirst = false;
			}
			
			read.setCigar(newCigar);
		}
	}
}
 

/**
 * Remove leading or trailing soft clips from the input read.
 * Does not modify a read entirely comprised of soft clips.
 */
public static void removeSoftClips(SAMRecord read) {
	
	Cigar cigar = read.getCigar();
	
	CigarElement firstElement = cigar.getCigarElement(0);
	CigarElement lastElement  = cigar.getCigarElement(cigar.numCigarElements()-1);
	
	if ((firstElement.getOperator() == CigarOperator.S) ||
		(lastElement.getOperator() == CigarOperator.S)) {
	
		Cigar newCigar = new Cigar();
		
		String bases = read.getReadString();
		//String qualities = read.getBaseQualityString();
				
		if (firstElement.getOperator() == CigarOperator.S) {
			bases = bases.substring(firstElement.getLength(), bases.length());
			//qualities = qualities.substring(firstElement.getLength(), qualities.length()-1);
		} else {
			newCigar.add(firstElement);
		}
		
		for (int i=1; i<cigar.numCigarElements()-1; i++) {
			newCigar.add(cigar.getCigarElement(i));
		}
		
		if (lastElement.getOperator() == CigarOperator.S) {
			bases = bases.substring(0, bases.length() - lastElement.getLength());
			//qualities = qualities.substring(0, qualities.length() - lastElement.getLength() - 1);
		} else {
			newCigar.add(lastElement);
		}
		
		read.setCigar(newCigar);
		read.setReadString(bases);
		//read.setBaseQualityString(qualities);
	}
}
 

@Test (groups = "unit" )
public void testRemoveSoftClips_withDeletionAndSoftClipAtEnd() {
	SAMRecord read = new SAMRecord(null);
	
	read.setReadName("TEST1");
	read.setCigarString("5M2D3M2S");
	read.setReadString("CCCCCCAGCC");
	
	SAMRecordUtils.removeSoftClips(read);
	
	Assert.assertEquals(read.getCigarString(), "5M2D3M");
	Assert.assertEquals(read.getReadString(), "CCCCCCAG");
}
 

@Test (groups = "unit" )
public void testRemoveSoftClips_withDeletionAndSoftClipAtStart() {
	SAMRecord read = new SAMRecord(null);
	
	read.setReadName("TEST1");
	read.setCigarString("2S5M2D3M");
	read.setReadString("CCCCCCAGCC");
	
	SAMRecordUtils.removeSoftClips(read);
	
	Assert.assertEquals(read.getCigarString(), "5M2D3M");
	Assert.assertEquals(read.getReadString(), "CCCCAGCC");
}
 

@NotNull
public static SAMRecord buildSamRecord(final int alignmentStart, @NotNull final String cigar, @NotNull final String readString,
        @NotNull final String qualities) {
    final SAMRecord record = new SAMRecord(null);
    record.setAlignmentStart(alignmentStart);
    record.setCigarString(cigar);
    record.setReadString(readString);
    record.setReadNegativeStrandFlag(false);
    record.setBaseQualityString(qualities);
    record.setMappingQuality(20);
    record.setDuplicateReadFlag(false);
    record.setReadUnmappedFlag(false);
    return record;
}
 

public void testCheckNhPicard() {
  final SAMRecord rec = new SAMRecord(null);
  rec.setReadString("TATTAGGATTGAGACTGGTAAAATGGNCCACCAAG");
  assertEquals(null, SamUtils.getNHOrIH(rec));
  rec.setAttribute("IH", 1);
  assertEquals(Integer.valueOf(1), SamUtils.getNHOrIH(rec));
  rec.setAttribute("NH", 2);
  assertEquals(Integer.valueOf(2), SamUtils.getNHOrIH(rec));
}
 

public void test() throws IOException {
  final SAMFileHeader header = new SAMFileHeader();
  header.setSortOrder(SAMFileHeader.SortOrder.coordinate);
  header.getSequenceDictionary().addSequence(new SAMSequenceRecord("seq", 1000000));
  final SAMRecord rec = new SAMRecord(header);
  rec.setAlignmentStart(500000);
  rec.setReferenceName("seq");
  rec.setReadName("read");
  rec.setReadString("acgt");
  rec.setBaseQualityString("####");
  rec.setCigarString("4=");
  try (TestDirectory dir = new TestDirectory()) {
    final MemoryPrintStream mps = new MemoryPrintStream();
    try (SamOutput out = SamOutput.getSamOutput(new File(dir, "foo"), mps.outputStream(), header, true, true, null)) {
      out.getWriter().addAlignment(rec);
    }
    mNano.check("samoutput_expected_1.sam", MainResult.run(new ExtractCli(), "--header", new File(dir, "foo.bam").getPath(), "seq:500000+1").out());
    assertTrue(new File(dir, "foo.bam.bai").exists());
    assertEquals("", mps.toString());

    try (SamOutput out = SamOutput.getSamOutput(new File(dir, "foo.sam"), mps.outputStream(), header, true, true, null)) {
      out.getWriter().addAlignment(rec);
    }
    mNano.check("samoutput_expected_1.sam", MainResult.run(new ExtractCli(), "--header", new File(dir, "foo.sam.gz").getPath(), "seq:500000+1").out());
    assertTrue(new File(dir, "foo.sam.gz.tbi").exists());
    assertEquals("", mps.toString());

    try (SamOutput out = SamOutput.getSamOutput(new File(dir, "foo.sam"), mps.outputStream(), header, false, true, null)) {
      out.getWriter().addAlignment(rec);
    }
    mNano.check("samoutput_expected_1.sam", FileHelper.fileToString(new File(dir, "foo.sam")));
    assertFalse(new File(dir, "foo.sam.tbi").exists());
    assertEquals("", mps.toString());

    try (SamOutput out = SamOutput.getSamOutput(new File("-"), mps.outputStream(), header, true, true, null)) {
      out.getWriter().addAlignment(rec);
    }
    mNano.check("samoutput_expected_1.sam", mps.toString());
  }
}
 

public void testFilterReadLength() {
  final SamFilterParams.SamFilterParamsBuilder builder = SamFilterParams.builder().minLength(10);
  final DefaultSamFilter f = new DefaultSamFilter(builder.create());
  final DefaultSamFilter notf = new DefaultSamFilter(builder.invertFilters(true).create());
  final SAMRecord rec = new SAMRecord(new SAMFileHeader());
  assertFalse(f.acceptRecord(rec));
  assertTrue(notf.acceptRecord(rec));
  rec.setReadString("atgcatgcatgcatgcatgcatgcatgc");
  assertTrue(f.acceptRecord(rec));
  assertFalse(notf.acceptRecord(rec));
}
 

@NotNull
static SAMRecord buildSamRecord(@NotNull final String cigar, @NotNull final String readString, @NotNull final String qualities) {
    final SAMRecord record = new SAMRecord(null);
    record.setAlignmentStart(1000);
    record.setCigarString(cigar);
    record.setReadString(readString);
    record.setReadNegativeStrandFlag(false);
    record.setBaseQualityString(qualities);
    record.setMappingQuality(20);
    record.setDuplicateReadFlag(false);
    record.setReadUnmappedFlag(false);
    return record;
}
 

/**
 * @return a 2-element array in which the first element is the unmapped SAM, and the second the mapped SAM.
 */
private File[] createSamFilesToBeMerged(final MultipleAlignmentSpec[] specs) {
    try {
        final File unmappedSam = File.createTempFile("unmapped.", ".sam");
        unmappedSam.deleteOnExit();
        final SAMFileWriterFactory factory = new SAMFileWriterFactory();
        final SAMFileHeader header = new SAMFileHeader();
        header.setSortOrder(SAMFileHeader.SortOrder.queryname);
        final SAMRecord unmappedRecord = new SAMRecord(header);

        unmappedRecord.setReadName("theRead");
        unmappedRecord.setReadString("ACGTACGTACGTACGT");
        unmappedRecord.setBaseQualityString("5555555555555555");
        unmappedRecord.setReadUnmappedFlag(true);

        final SAMFileWriter unmappedWriter = factory.makeSAMWriter(header, false, unmappedSam);
        unmappedWriter.addAlignment(unmappedRecord);
        unmappedWriter.close();

        final File alignedSam = File.createTempFile("aligned.", ".sam");
        alignedSam.deleteOnExit();

        final String sequence = "chr1";
        // Populate the header with SAMSequenceRecords
        header.setSequenceDictionary(SAMSequenceDictionaryExtractor.extractDictionary(sequenceDict2.toPath()));

        final SAMFileWriter alignedWriter = factory.makeSAMWriter(header, false, alignedSam);
        for (final MultipleAlignmentSpec spec : specs) {
            final SAMRecord alignedRecord = new SAMRecord(header);
            alignedRecord.setReadName(unmappedRecord.getReadName());
            alignedRecord.setReadBases(unmappedRecord.getReadBases());
            alignedRecord.setBaseQualities(unmappedRecord.getBaseQualities());
            alignedRecord.setReferenceName(sequence);
            alignedRecord.setAlignmentStart(1);
            alignedRecord.setReadNegativeStrandFlag(spec.reverseStrand);
            alignedRecord.setCigarString(spec.cigar);
            alignedRecord.setMappingQuality(spec.mapQ);
            if (spec.oneOfTheBest) {
                alignedRecord.setAttribute(ONE_OF_THE_BEST_TAG, 1);
            }
            alignedWriter.addAlignment(alignedRecord);
        }
        alignedWriter.close();

        return new File[]{unmappedSam, alignedSam};
    } catch (IOException e) {
        throw new PicardException(e.getMessage(), e);
    }
}
 

private void testBestFragmentMapqStrategy(final String testName, final int[] firstMapQs, final int[] secondMapQs,
                                          final boolean includeSecondary, final int expectedFirstMapq,
                                          final int expectedSecondMapq) throws Exception {
    final File unmappedSam = File.createTempFile("unmapped.", ".sam");
    unmappedSam.deleteOnExit();
    final SAMFileWriterFactory factory = new SAMFileWriterFactory();
    final SAMFileHeader header = new SAMFileHeader();
    header.setSortOrder(SAMFileHeader.SortOrder.queryname);

    final String readName = "theRead";
    final SAMRecord firstUnmappedRead = new SAMRecord(header);
    firstUnmappedRead.setReadName(readName);
    firstUnmappedRead.setReadString("ACGTACGTACGTACGT");
    firstUnmappedRead.setBaseQualityString("5555555555555555");
    firstUnmappedRead.setReadUnmappedFlag(true);
    firstUnmappedRead.setMateUnmappedFlag(true);
    firstUnmappedRead.setReadPairedFlag(true);
    firstUnmappedRead.setFirstOfPairFlag(true);

    final SAMRecord secondUnmappedRead = new SAMRecord(header);
    secondUnmappedRead.setReadName(readName);
    secondUnmappedRead.setReadString("TCGAACGTTCGAACTG");
    secondUnmappedRead.setBaseQualityString("6666666666666666");
    secondUnmappedRead.setReadUnmappedFlag(true);
    secondUnmappedRead.setMateUnmappedFlag(true);
    secondUnmappedRead.setReadPairedFlag(true);
    secondUnmappedRead.setSecondOfPairFlag(true);

    final SAMFileWriter unmappedWriter = factory.makeSAMWriter(header, false, unmappedSam);
    unmappedWriter.addAlignment(firstUnmappedRead);
    unmappedWriter.addAlignment(secondUnmappedRead);
    unmappedWriter.close();

    final File alignedSam = File.createTempFile("aligned.", ".sam");
    alignedSam.deleteOnExit();

    final String sequence = "chr1";
    // Populate the header with SAMSequenceRecords
    header.setSequenceDictionary(SAMSequenceDictionaryExtractor.extractDictionary(sequenceDict2.toPath()));

    final SAMFileWriter alignedWriter = factory.makeSAMWriter(header, false, alignedSam);

    addAlignmentsForBestFragmentMapqStrategy(alignedWriter, firstUnmappedRead, sequence, firstMapQs);
    addAlignmentsForBestFragmentMapqStrategy(alignedWriter, secondUnmappedRead, sequence, secondMapQs);
    alignedWriter.close();

    final File output = File.createTempFile("testBestFragmentMapqStrategy." + testName, ".sam");
    output.deleteOnExit();
    doMergeAlignment(unmappedSam, Collections.singletonList(alignedSam),
            null, null, null, null,
            false, true, false, 1,
            "0", "1.0", "align!", "myAligner",
            true,
            fasta, output,
            SamPairUtil.PairOrientation.FR,
            MergeBamAlignment.PrimaryAlignmentStrategy.BestEndMapq,
            null, includeSecondary, null, null);
    final SamReader reader = SamReaderFactory.makeDefault().open(output);

    int numFirstRecords = 0;
    int numSecondRecords = 0;
    int firstPrimaryMapq = -1;
    int secondPrimaryMapq = -1;
    for (final SAMRecord rec: reader) {
        Assert.assertTrue(rec.getReadPairedFlag());
        if (rec.getFirstOfPairFlag()) ++numFirstRecords;
        else if (rec.getSecondOfPairFlag()) ++ numSecondRecords;
        else Assert.fail("unpossible!");
        if (!rec.getReadUnmappedFlag() && !rec.getNotPrimaryAlignmentFlag()) {
            if (rec.getFirstOfPairFlag()) {
                Assert.assertEquals(firstPrimaryMapq, -1);
                firstPrimaryMapq = rec.getMappingQuality();
            } else {
                Assert.assertEquals(secondPrimaryMapq, -1);
                secondPrimaryMapq = rec.getMappingQuality();
            }
        } else if (rec.getNotPrimaryAlignmentFlag()) {
            Assert.assertTrue(rec.getMateUnmappedFlag());
        }
    }
    reader.close();
    Assert.assertEquals(firstPrimaryMapq, expectedFirstMapq);
    Assert.assertEquals(secondPrimaryMapq, expectedSecondMapq);
    if (!includeSecondary) {
        Assert.assertEquals(numFirstRecords, 1);
        Assert.assertEquals(numSecondRecords, 1);
    } else {
        // If no alignments for an end, there will be a single unmapped record
        Assert.assertEquals(numFirstRecords, Math.max(1, firstMapQs.length));
        Assert.assertEquals(numSecondRecords, Math.max(1, secondMapQs.length));
    }
}
 

/**
 * Confirm that paired reads are rejected by PrimaryAlignmentStrategy.EarliestFragment.
 */
@Test(expectedExceptions = UnsupportedOperationException.class)
public void testEarliestFragmentStrategyPaired() throws Exception {
    final File output = File.createTempFile("mergeTest", ".sam");
    output.deleteOnExit();

    final File unmappedSam = File.createTempFile("unmapped.", ".sam");
    unmappedSam.deleteOnExit();
    final SAMFileWriterFactory factory = new SAMFileWriterFactory();
    final SAMFileHeader header = new SAMFileHeader();
    header.setSortOrder(SAMFileHeader.SortOrder.queryname);
    final String cigar = "16M";

    final SAMRecord firstOfPair = new SAMRecord(header);
    firstOfPair.setReadName("theRead");
    firstOfPair.setReadString("ACGTACGTACGTACGT");
    firstOfPair.setBaseQualityString("5555555555555555");
    firstOfPair.setReadUnmappedFlag(true);
    firstOfPair.setReadPairedFlag(true);
    firstOfPair.setFirstOfPairFlag(true);

    final SAMRecord secondOfPair = new SAMRecord(header);
    secondOfPair.setReadName("theRead");
    secondOfPair.setReadString("ACGTACGTACGTACGT");
    secondOfPair.setBaseQualityString("5555555555555555");
    secondOfPair.setReadUnmappedFlag(true);
    secondOfPair.setReadPairedFlag(true);
    secondOfPair.setSecondOfPairFlag(true);
    SamPairUtil.setMateInfo(firstOfPair, secondOfPair);

    final SAMFileWriter unmappedWriter = factory.makeSAMWriter(header, false, unmappedSam);
    unmappedWriter.addAlignment(firstOfPair);
    unmappedWriter.addAlignment(secondOfPair);
    unmappedWriter.close();

    final File alignedSam = File.createTempFile("aligned.", ".sam");
    alignedSam.deleteOnExit();

    // Populate the header with SAMSequenceRecords
    header.setSequenceDictionary(SAMSequenceDictionaryExtractor.extractDictionary(sequenceDict2.toPath()));

    // Create 2 alignments for each end of pair
    final SAMFileWriter alignedWriter = factory.makeSAMWriter(header, false, alignedSam);
    for (int i = 1; i <= 2; ++i) {
        final SAMRecord firstOfPairAligned = new SAMRecord(header);
        firstOfPairAligned.setReadName(firstOfPair.getReadName());
        firstOfPairAligned.setReadBases(firstOfPair.getReadBases());
        firstOfPairAligned.setBaseQualities(firstOfPair.getBaseQualities());
        firstOfPairAligned.setReferenceName("chr1");
        firstOfPairAligned.setAlignmentStart(i);
        firstOfPairAligned.setCigarString(cigar);
        firstOfPairAligned.setMappingQuality(100);
        firstOfPairAligned.setReadPairedFlag(true);
        firstOfPairAligned.setFirstOfPairFlag(true);
        firstOfPairAligned.setAttribute(SAMTag.HI.name(), i);

        final SAMRecord secondOfPairAligned = new SAMRecord(header);
        secondOfPairAligned.setReadName(secondOfPair.getReadName());
        secondOfPairAligned.setReadBases(secondOfPair.getReadBases());
        secondOfPairAligned.setBaseQualities(secondOfPair.getBaseQualities());
        secondOfPairAligned.setReferenceName("chr1");
        secondOfPairAligned.setAlignmentStart(i + 10);
        secondOfPairAligned.setCigarString(cigar);
        secondOfPairAligned.setMappingQuality(100);
        secondOfPairAligned.setReadPairedFlag(true);
        secondOfPairAligned.setSecondOfPairFlag(true);
        secondOfPairAligned.setAttribute(SAMTag.HI.name(), i);

        SamPairUtil.setMateInfo(firstOfPairAligned, secondOfPairAligned);

        alignedWriter.addAlignment(firstOfPairAligned);
        alignedWriter.addAlignment(secondOfPairAligned);
    }
    alignedWriter.close();

    doMergeAlignment(unmappedSam, Collections.singletonList(alignedSam),
            null, null, null, null,
            false, true, false, 1,
            "0", "1.0", "align!", "myAligner",
            true,
            fasta, output,
            SamPairUtil.PairOrientation.FR,
            MergeBamAlignment.PrimaryAlignmentStrategy.EarliestFragment,
            null, null, null, null);

    Assert.fail("Exception was not thrown");
}
 

/**
 * Takes an individual SAMRecord and applies the set of changes/reversions to it that
 * have been requested by program level options.
 */
public void revertSamRecord(final SAMRecord rec) {
    if (RESTORE_ORIGINAL_QUALITIES) {
        final byte[] oq = rec.getOriginalBaseQualities();
        if (oq != null) {
            rec.setBaseQualities(oq);
            rec.setOriginalBaseQualities(null);
        }
    }

    if (REMOVE_DUPLICATE_INFORMATION) {
        rec.setDuplicateReadFlag(false);
    }

    if (REMOVE_ALIGNMENT_INFORMATION) {
        if (rec.getReadNegativeStrandFlag()) {
            rec.reverseComplement(true);
            rec.setReadNegativeStrandFlag(false);
        }

        // Remove all alignment based information about the read itself
        rec.setReferenceIndex(SAMRecord.NO_ALIGNMENT_REFERENCE_INDEX);
        rec.setAlignmentStart(SAMRecord.NO_ALIGNMENT_START);
        rec.setCigarString(SAMRecord.NO_ALIGNMENT_CIGAR);
        rec.setMappingQuality(SAMRecord.NO_MAPPING_QUALITY);

        rec.setInferredInsertSize(0);
        rec.setNotPrimaryAlignmentFlag(false);
        rec.setProperPairFlag(false);
        rec.setReadUnmappedFlag(true);

        // Then remove any mate flags and info related to alignment
        rec.setMateAlignmentStart(SAMRecord.NO_ALIGNMENT_START);
        rec.setMateNegativeStrandFlag(false);
        rec.setMateReferenceIndex(SAMRecord.NO_ALIGNMENT_REFERENCE_INDEX);
        rec.setMateUnmappedFlag(rec.getReadPairedFlag());

        if (RESTORE_HARDCLIPS) {
            String hardClippedBases = rec.getStringAttribute(AbstractAlignmentMerger.HARD_CLIPPED_BASES_TAG);
            String hardClippedQualities = rec.getStringAttribute(AbstractAlignmentMerger.HARD_CLIPPED_BASE_QUALITIES_TAG);
            if (hardClippedBases != null && hardClippedQualities != null) {
                // Record has already been reverse complemented if this was on the negative strand
                rec.setReadString(rec.getReadString() + hardClippedBases);
                rec.setBaseQualities(SAMUtils.fastqToPhred(SAMUtils.phredToFastq(rec.getBaseQualities()) + hardClippedQualities));

                // Remove hard clipping storage tags
                rec.setAttribute(AbstractAlignmentMerger.HARD_CLIPPED_BASES_TAG, null);
                rec.setAttribute(AbstractAlignmentMerger.HARD_CLIPPED_BASE_QUALITIES_TAG, null);
            }
        }

        // And then remove any tags that are calculated from the alignment
        ATTRIBUTE_TO_CLEAR.forEach(tag -> rec.setAttribute(tag, null));
    }
}
 

/**
 * Confirm that after the requisite number of sightings of adapter, the list is trimmed
 */
@Test(dataProvider = "testAdapterListTruncationDataProvider")
public void testAdapterListTruncation(final IlluminaAdapterPair adapterPair) {
    final int thresholdForTruncatingAdapterList = 20;
    final int readLength = 100;

    // Throw all the adapter pairs into the marker.  There is some danger in doing this because
    // IlluminaAdapterPair.ALTERNATIVE_SINGLE_END has 3' that is a substring of some of the others.  In the enum it appears
    // last so it is tested last.
    final AdapterMarker marker =
            new AdapterMarker(IlluminaUtil.IlluminaAdapterPair.values()).
                    setThresholdForSelectingAdaptersToKeep(thresholdForTruncatingAdapterList);
    int adapterPosition = 1;

    Assert.assertTrue(adapterPosition + thresholdForTruncatingAdapterList < readLength - ClippingUtility.MIN_MATCH_BASES,
            "Test is configured improperly -- eventually will not be enough adapter bases in read.");

    int originalNumberOfAdapters = marker.getAdapters().length;
    for (int i = 0; i < thresholdForTruncatingAdapterList; ++i) {

        // First, a read with no adapter in it.
        SAMRecord rec = new SAMRecord(null);
        rec.setReadString(makeBogusReadString(readLength));
        AdapterPair matchedPair = marker.adapterTrimIlluminaSingleRead(rec);
        Assert.assertNull(matchedPair);
        Assert.assertNull(rec.getAttribute(ReservedTagConstants.XT));

        // Adapter list should not be truncated yet
        Assert.assertEquals(marker.getAdapters().length, originalNumberOfAdapters);

        // Then, a record with some adapter sequence in it.
        rec = createSamRecordWithAdapterSequence(readLength, adapterPair, adapterPosition);
        matchedPair = marker.adapterTrimIlluminaSingleRead(rec);
        Assert.assertNotNull(matchedPair);
        Assert.assertEquals(rec.getIntegerAttribute(ReservedTagConstants.XT).intValue(), adapterPosition + 1,
                rec.getReadString() + " matched " + matchedPair);

        // Put adapter in different place next time.
        adapterPosition++;
    }

    Assert.assertEquals(marker.getAdapters().length, 1, "Did not truncate adapter list to 1 element");
    Assert.assertTrue(marker.getAdapters()[0].getName().contains(adapterPair.getName()),
            String.format("Expected '%s' to contain '%s'", marker.getAdapters()[0].getName(), adapterPair.getName()));
}