Fix for Adam K's reported bug: we weren't handling reads that were entirely insertions properly in LIBS. Specifically, the event bases were off-by-one (which was disasterous in Adam's case with a 1bp read). Added a unit test to cover this case.
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@ -301,6 +301,7 @@ public class LocusIteratorByState extends LocusIterator {
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final CigarOperator op = state.getCurrentCigarOperator(); // current cigar operator
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final CigarElement nextElement = state.peekForwardOnGenome(); // next cigar element
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final CigarElement lastElement = state.peekBackwardOnGenome(); // last cigar element
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final boolean isSingleElementCigar = nextElement == lastElement;
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final CigarOperator nextOp = nextElement.getOperator(); // next cigar operator
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final CigarOperator lastOp = lastElement.getOperator(); // last cigar operator
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final int readOffset = state.getReadOffset(); // the base offset on this read
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@ -308,7 +309,7 @@ public class LocusIteratorByState extends LocusIterator {
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final boolean isBeforeDeletion = nextOp == CigarOperator.DELETION;
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final boolean isAfterDeletion = lastOp == CigarOperator.DELETION;
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final boolean isBeforeInsertion = nextOp == CigarOperator.INSERTION;
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final boolean isAfterInsertion = lastOp == CigarOperator.INSERTION;
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final boolean isAfterInsertion = lastOp == CigarOperator.INSERTION && !isSingleElementCigar;
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final boolean isNextToSoftClip = nextOp == CigarOperator.S || (state.getGenomeOffset() == 0 && read.getSoftStart() != read.getAlignmentStart());
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int nextElementLength = nextElement.getLength();
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@ -328,8 +329,10 @@ public class LocusIteratorByState extends LocusIterator {
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else {
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if (!filterBaseInRead(read, location.getStart())) {
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String insertedBaseString = null;
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if (nextOp == CigarOperator.I)
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insertedBaseString = new String(Arrays.copyOfRange(read.getReadBases(), readOffset + 1, readOffset + 1 + nextElement.getLength()));
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if (nextOp == CigarOperator.I) {
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final int insertionOffset = isSingleElementCigar ? 0 : 1;
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insertedBaseString = new String(Arrays.copyOfRange(read.getReadBases(), readOffset + insertionOffset, readOffset + insertionOffset + nextElement.getLength()));
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}
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pile.add(new PileupElement(read, readOffset, false, isBeforeDeletion, isAfterDeletion, isBeforeInsertion, isAfterInsertion, isNextToSoftClip, insertedBaseString, nextElementLength));
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size++;
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@ -104,7 +104,7 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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after.setBaseQualities(new byte[] {20,20,20,20,20,20,20,20,20,20});
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after.setCigarString("10M");
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List<SAMRecord> reads = Arrays.asList(before,during,after);
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List<SAMRecord> reads = Arrays.asList(before, during, after);
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// create the iterator by state with the fake reads and fake records
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li = makeLTBS(reads,readAttributes);
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@ -134,9 +134,9 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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// create a test version of the Reads object
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ReadProperties readAttributes = createTestReadProperties();
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SAMRecord indelOnlyRead = ArtificialSAMUtils.createArtificialRead(header,"indelOnly",0,firstLocus,76);
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SAMRecord indelOnlyRead = ArtificialSAMUtils.createArtificialRead(header, "indelOnly", 0, firstLocus, 76);
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indelOnlyRead.setReadBases(Utils.dupBytes((byte)'A',76));
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indelOnlyRead.setBaseQualities(Utils.dupBytes((byte)'@',76));
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indelOnlyRead.setBaseQualities(Utils.dupBytes((byte) '@', 76));
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indelOnlyRead.setCigarString("76I");
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List<SAMRecord> reads = Arrays.asList(indelOnlyRead);
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@ -148,10 +148,10 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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// and considers it to be an indel-containing read.
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Assert.assertTrue(li.hasNext(),"Should have found a whole-indel read in the normal base pileup without extended events enabled");
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AlignmentContext alignmentContext = li.next();
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Assert.assertEquals(alignmentContext.getLocation().getStart(),firstLocus,"Base pileup is at incorrect location.");
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Assert.assertEquals(alignmentContext.getLocation().getStart(), firstLocus, "Base pileup is at incorrect location.");
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ReadBackedPileup basePileup = alignmentContext.getBasePileup();
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Assert.assertEquals(basePileup.getReads().size(),1,"Pileup is of incorrect size");
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Assert.assertSame(basePileup.getReads().get(0),indelOnlyRead,"Read in pileup is incorrect");
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Assert.assertSame(basePileup.getReads().get(0), indelOnlyRead, "Read in pileup is incorrect");
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}
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/**
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@ -168,7 +168,7 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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leadingRead.setCigarString("1M75I");
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SAMRecord indelOnlyRead = ArtificialSAMUtils.createArtificialRead(header,"indelOnly",0,secondLocus,76);
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indelOnlyRead.setReadBases(Utils.dupBytes((byte)'A',76));
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indelOnlyRead.setReadBases(Utils.dupBytes((byte) 'A', 76));
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indelOnlyRead.setBaseQualities(Utils.dupBytes((byte)'@',76));
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indelOnlyRead.setCigarString("76I");
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@ -177,7 +177,7 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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fullMatchAfterIndel.setBaseQualities(Utils.dupBytes((byte)'@',76));
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fullMatchAfterIndel.setCigarString("75I1M");
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List<SAMRecord> reads = Arrays.asList(leadingRead,indelOnlyRead,fullMatchAfterIndel);
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List<SAMRecord> reads = Arrays.asList(leadingRead, indelOnlyRead, fullMatchAfterIndel);
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// create the iterator by state with the fake reads and fake records
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li = makeLTBS(reads, createTestReadProperties());
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@ -204,7 +204,36 @@ public class LocusIteratorByStateUnitTest extends BaseTest {
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numAlignmentContextsFound++;
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}
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Assert.assertEquals(numAlignmentContextsFound,2,"Found incorrect number of alignment contexts");
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Assert.assertEquals(numAlignmentContextsFound, 2, "Found incorrect number of alignment contexts");
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}
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/**
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* Test to make sure that reads supporting only an indel (example cigar string: 76I) do
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* not negatively influence the ordering of the pileup.
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*/
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@Test
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public void testWholeIndelReadTest2() {
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final int firstLocus = 44367788, secondLocus = firstLocus + 1;
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SAMRecord read = ArtificialSAMUtils.createArtificialRead(header,"read",0,secondLocus,1);
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read.setReadBases(Utils.dupBytes((byte) 'A', 1));
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read.setBaseQualities(Utils.dupBytes((byte) '@', 1));
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read.setCigarString("1I");
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List<SAMRecord> reads = Arrays.asList(read);
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// create the iterator by state with the fake reads and fake records
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li = makeLTBS(reads, createTestReadProperties());
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while(li.hasNext()) {
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AlignmentContext alignmentContext = li.next();
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ReadBackedPileup p = alignmentContext.getBasePileup();
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Assert.assertTrue(p.getNumberOfElements() == 1);
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PileupElement pe = p.iterator().next();
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Assert.assertTrue(pe.isBeforeInsertion());
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Assert.assertFalse(pe.isAfterInsertion());
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Assert.assertEquals(pe.getEventBases(), "A");
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}
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}
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private static ReadProperties createTestReadProperties() {
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