public void TryStitch_NoXC_Stitchable()
{
//Reads without XC tags that do overlap should be added as one merged read in basic stitcher
var basicStitcher = StitcherTestHelpers.GetStitcher(10);
var alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
basicStitcher.TryStitch(alignmentSet);
Assert.Equal(1, alignmentSet.ReadsForProcessing.Count);
}
public void TryStitch_NoXC_Stitchable()
{
var xcStitcherXcRequired = GetXCStitcher();
//Reads without XC tags that do overlap should be added separately in XC stitcher when xc is required
var alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
StitcherTestHelpers.TryStitchAndAssertFailed(xcStitcherXcRequired, alignmentSet);
}
public void TryStitch_NoXC_Unstitchable()
{
var read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12345, new CigarAlignment("8M"), qualityForAll: 30);
var read2_noOverlap = TestHelper.CreateRead("chr1", "A", 2384, new CigarAlignment("1M"), qualityForAll: 30);
var read2_overlap = TestHelper.CreateRead("chr1", "ATCGTT", 12349, new CigarAlignment("6M"), qualityForAll: 30);
var read2_diffChrom = TestHelper.CreateRead("chr2", "ATCGTT", 12349, new CigarAlignment("6M"), qualityForAll: 30);
var read2_nonOverlap_border = TestHelper.CreateRead("chr1", "AT", 12343, new CigarAlignment("2M"), qualityForAll: 30);
var stitcher = StitcherTestHelpers.GetStitcher(10, true);
// -----------------------------------------------
// Either of the partner reads is missing*
// *(only read that could be missing is read 2, if read 1 was missing couldn't create alignment set)
// -----------------------------------------------
// Should throw an exception
var alignmentSet = new AlignmentSet(read1, null);
Assert.Throws <ArgumentException>(() => stitcher.TryStitch(alignmentSet));
// -----------------------------------------------
// No overlap, reads are far away
// -----------------------------------------------
// Shouldn't stitch
alignmentSet = new AlignmentSet(read1, read2_noOverlap);
StitcherTestHelpers.TryStitchAndAssertFailed(stitcher, alignmentSet);
// -----------------------------------------------
// No overlap, reads are directly neighboring
// -----------------------------------------------
// Shouldn't stitch
alignmentSet = new AlignmentSet(read1, read2_nonOverlap_border);
StitcherTestHelpers.TryStitchAndAssertFailed(stitcher, alignmentSet);
// -----------------------------------------------
// No overlap, reads on diff chromosomes
// -----------------------------------------------
// Should throw exception
alignmentSet = new AlignmentSet(read1, read2_diffChrom);
var ex = Assert.Throws <ArgumentException>(() => stitcher.TryStitch(alignmentSet));
Assert.Contains("Partner reads are from different chromosomes", ex.Message, StringComparison.InvariantCultureIgnoreCase); // This is brittle but since a variety of exceptions can happen in this process want to make sure it's this specific one
}
public void TryStitch_CalculateStitchedCigar()
{
// -----------------------------------------------
// Read position maps disagree
// -----------------------------------------------
// Should throw out the pair
var read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12345,
new CigarAlignment("2M2D3M1D3M"), qualityForAll: 30); //Within the overlap, we have a deletion so there will be a shifting of positions from that point on
var read2 = TestHelper.CreateRead("chr1", "ATCGATCG", 12349,
new CigarAlignment("8M"), qualityForAll: 30);
var stitcher = StitcherTestHelpers.GetStitcher(10);
var alignmentSet = new AlignmentSet(read1, read2);
Assert.True(!alignmentSet.ReadsForProcessing.Any());
// -----------------------------------------------
// When calculating stitched cigar, stitched cigar should have
// - everything from read1 before the overlap
// - everything from read2 starting from the overlap
// But since we ensure that the position maps agree in the overlap region, it's really not a matter of one taking precedence over the other
// 1234... 1 - - 2 3 4 5 6 - - 7 8 9 0
// Read1 X X X X X X X X - - - - -
// Read1 M I I M M M M M - - - - -
// Read2 - - - X X X X X X X X - -
// Read2 - - - M M M M M I M M - -
// -----------------------------------------------
// Stitched cigar should have R1's insertion from before the overlap and R2's insertion from after the overlap
read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12341,
new CigarAlignment("1M2I5M"), qualityForAll: 30);
read2 = TestHelper.CreateRead("chr1", "ATCGATCG", 12342,
new CigarAlignment("5M1I2M"), qualityForAll: 30);
stitcher = StitcherTestHelpers.GetStitcher(10);
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
Assert.Equal("1M2I5M1I2M", StitcherTestHelpers.GetMergedRead(alignmentSet).CigarData.ToString());
}
public static void TestSuccesfullyStitchedRead(Read read1, Read read2, int minQscore, string xcTag, Action <Read> assertions)
{
var alignmentSet = new AlignmentSet(read1, read2);
var stitcher = StitcherTestHelpers.GetStitcher(minQscore);
stitcher.TryStitch(alignmentSet);
// -----------------------------------------------
// Basic Stitcher, No XC tag
// -----------------------------------------------
CheckMergedRead(xcTag, assertions, alignmentSet);
// -----------------------------------------------
// XC Stitcher (XC Req), No XC tag
// -----------------------------------------------
alignmentSet = new AlignmentSet(read1, read2);
stitcher = StitcherTestHelpers.GetStitcher(minQscore, true);
StitcherTestHelpers.TryStitchAndAssertFailed(stitcher, alignmentSet);
// -----------------------------------------------
// Basic Stitcher, With XC tag
// -----------------------------------------------
read1.StitchedCigar = new CigarAlignment(xcTag);
read2.StitchedCigar = new CigarAlignment(xcTag);
alignmentSet = new AlignmentSet(read1, read2);
stitcher = StitcherTestHelpers.GetStitcher(minQscore);
stitcher.TryStitch(alignmentSet);
CheckMergedRead(xcTag, assertions, alignmentSet);
// -----------------------------------------------
// XC Stitcher (XC Req), With XC tag
// -----------------------------------------------
alignmentSet = new AlignmentSet(read1, read2);
stitcher = StitcherTestHelpers.GetStitcher(minQscore, true);
stitcher.TryStitch(alignmentSet);
CheckMergedRead(xcTag, assertions, alignmentSet);
}
public void TryStitch_CalculateStitchedCigar()
{
// -----------------------------------------------
// Read position maps disagree
// -----------------------------------------------
var read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12345, new CigarAlignment("2M2D3M1D3M"), qualityForAll: 30);
var read2 = TestHelper.CreateRead("chr1", "ATCGATCG", 12349, new CigarAlignment("8M"), qualityForAll: 30);
// [If require XC]
// We never calculate stitched cigar anyway. Bounce out to processing separately.
var stitcher = GetXCStitcher();
var alignmentSet = new AlignmentSet(read1, read2);
StitcherTestHelpers.TryStitchAndAssertFailed(stitcher, alignmentSet);
// -----------------------------------------------
// When calculating stitched cigar, stitched cigar should have
// - everything from read1 before the overlap
// - everything from read2 starting from the overlap
// But since we ensure that the position maps agree in the overlap region, it's really not a matter of one taking precedence over the other
// 1234... 1 - - 2 3 4 5 6 - - 7 8 9 0
// Read1 X X X X X X X X - - - - -
// Read1 M I I M M M M M - - - - -
// Read2 - - - X X X X X X X X - -
// Read2 - - - M M M M M I M M - -
// -----------------------------------------------
// [If require XC]
// No stitched cigar, and separate reads
read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12341, new CigarAlignment("1M2I5M"), qualityForAll: 30);
read2 = TestHelper.CreateRead("chr1", "ATCGATCG", 12342, new CigarAlignment("5M1I2M"), qualityForAll: 30);
stitcher = GetXCStitcher();
alignmentSet = new AlignmentSet(read1, read2);
StitcherTestHelpers.TryStitchAndAssertFailed(stitcher, alignmentSet);
}
public void TryStitch_SoftclipScenarios()
{
//Migrated from old Pisces: Originally called CallSomaticVariants_MergeRead2First
var sequence =
"GGGGCCACGCGGGGAGCAGCCTCTGGCATTCTGGGAGCTTCATCTGGACCTGGGTCTTCAGTGAACCATTGTTCAATATCGTCCGGGGACAGCATCAAATCATCCATTGCTTGGGACGGCAAGGGGGACTGTAGATGGGTGAAAAGAGCA";
var read1 = TestHelper.CreateRead("chr1",
sequence,
7579464,
new CigarAlignment("2S122M26S"),
Enumerable.Repeat((byte)30, sequence.Length).ToArray(),
7579464);
StitcherTestHelpers.SetReadDirections(read1, DirectionType.Forward);
sequence =
"GTGTAGGAGCTGCTGGTGCAGGGGCCACGCGGGGAGCAGCCTCTGGCATTCTGGGAGCTTCATCTGGACCTGGGTCTTCAGTGAACAATTGTTCAATATCGTCCGGGGCCAGCATCAAATCATCCATTGCTTGGGACGGCAAGGGGGACT";
var read2 = TestHelper.CreateRead("chr1",
sequence,
7579464,
new CigarAlignment("22S122M6S"),
Enumerable.Repeat((byte)30, sequence.Length).ToArray(),
7579464);
StitcherTestHelpers.SetReadDirections(read2, DirectionType.Reverse);
string expected =
"GTGTAGGAGCTGCTGGTGCAGGGGCCACGCGGGGAGCAGCCTCTGGCATTCTGGGAGCTTCATCTGGACCTGGGTCTTCAGTGAACNATTGTTCAATATCGTCCGGGGNCAGCATCAAATCATCCATTGCTTGGGACGGCAAGGGGGACTGTAGATGGGTGAAAAGAGCA";
// both reads have the same reference position, but read2 really starts earlier
// make sure we behave properly
TestSuccesfullyStitchedRead(read1, read2, 0, "22S122M26S", (mergedRead) =>
{
Assert.NotNull(mergedRead);
Assert.Equal(mergedRead.Sequence, expected);
});
}
public void TryStitch_ConsensusSequence()
{
// 1234... 1 - - 2 3 4 5 6 - - 7 8 9 0 //Reference Positions
// Read1 X X X X X X X X - - - - -
// Read1 M I I M M M M M - - - - -
// Read1 A T C G A T C G - - - - -
// Read2 - - - X X X X X X X X - -
// Read2 - - - M M M M M I M M - -
// Read2 - - - A T C G A T C G - -
var r1qualities = 30;
var r2qualities = 20;
var read1 = TestHelper.CreateRead("chr1", "TTTTTTTT", 12341,
new CigarAlignment("1M2I5M"), qualityForAll: (byte)r1qualities);
var read2 = TestHelper.CreateRead("chr1", "AAAAAAAA", 12342,
new CigarAlignment("5M1I2M"), qualityForAll: (byte)r2qualities);
var stitcher = StitcherTestHelpers.GetStitcher(10);
var alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
// Merged A T C ? ? ? ? ? T C G - -
// Merged M I I M M M M M I M M - -
// Merged 0 1 2 3 4 5 6 7 8 9 0 1 2
var overlapStart = 3;
var overlapEnd = 8;
var overlapLength = 5;
//Consensus sequence should have everything from read1 for positions before overlap
var mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal("TTT", mergedRead.Sequence.Substring(0, overlapStart));
//Consensus sequence should have everything from read2 for positions after overlap
Assert.Equal("AAA", mergedRead.Sequence.Substring(overlapEnd, 3));
//Consensus sequence should have an N where we have two high-quality (both above min) disagreeing bases
Assert.Equal("NNNNN", mergedRead.Sequence.Substring(overlapStart, 5));
//Consensus sequence should have 0 quality where we have two high-quality (both above min) disagreeing bases
Assert.True(mergedRead.Qualities.Take(overlapStart).All(q => q == r1qualities));
Assert.True(mergedRead.Qualities.Skip(overlapStart).Take(overlapLength).All(q => q == 0));
Assert.True(mergedRead.Qualities.Skip(overlapEnd).Take(mergedRead.Sequence.Length - overlapEnd).All(q => q == r2qualities));
//Consensus sequence should take higher quality base if one or more of the bases is below min quality
//Read 2 trumps whole overlap
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 5, 5, 5, 5, 5 };
read2.BamAlignment.Qualities = new byte[] { 40, 40, 40, 40, 40, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(read2.Sequence.Substring(0, 5), mergedRead.Sequence.Substring(overlapStart, 5));
Assert.Equal("TTTAAAAAAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 40, 40, 40, 40, 40, 20, 19, 18 }, mergedRead.Qualities);
//Read 1 trumps whole overlap
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 40, 40, 40, 40, 40 };
read2.BamAlignment.Qualities = new byte[] { 5, 5, 5, 5, 5, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(read1.Sequence.Substring(3, 5), mergedRead.Sequence.Substring(overlapStart, 5));
Assert.Equal("TTTTTTTTAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 40, 40, 40, 40, 40, 20, 19, 18 }, mergedRead.Qualities);
//Little bit of each
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 5, 45, 5, 45, 5 };
read2.BamAlignment.Qualities = new byte[] { 40, 5, 40, 5, 40, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal("TTTATATAAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 40, 45, 40, 45, 40, 20, 19, 18 }, mergedRead.Qualities);
//Consensus sequence should take base and assign the higher quality if both bases agree
var read2_agreeingBases = TestHelper.CreateRead("chr1", "TTTTTTTT", 12342,
new CigarAlignment("5M1I2M"), new byte[] { 40, 5, 40, 5, 40, 20, 19, 18 });
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 5, 45, 5, 45, 5 };
alignmentSet = new AlignmentSet(read1, read2_agreeingBases);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal("TTTTTTTTTTT", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 40, 45, 40, 45, 40, 20, 19, 18 }, mergedRead.Qualities);
//Bases disagree and both are below minimum quality, read1>read2 : take base/q from read1
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 8, 8, 8, 8, 8 };
read2.BamAlignment.Qualities = new byte[] { 5, 5, 5, 5, 5, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(read1.Sequence.Substring(3, 5), mergedRead.Sequence.Substring(overlapStart, 5));
Assert.Equal("TTTTTTTTAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 8, 8, 8, 8, 8, 20, 19, 18 }, mergedRead.Qualities);
//Bases disagree and both are below minimum quality, read2>read1 : take base/q from read2
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 5, 5, 5, 5, 5 };
read2.BamAlignment.Qualities = new byte[] { 8, 8, 8, 8, 8, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(read2.Sequence.Substring(0, 5), mergedRead.Sequence.Substring(overlapStart, 5));
Assert.Equal("TTTAAAAAAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 8, 8, 8, 8, 8, 20, 19, 18 }, mergedRead.Qualities);
//Bases disagree and both are below minimum quality, read1==read2 : take base/q from read1
read1.BamAlignment.Qualities = new byte[] { 30, 30, 30, 5, 5, 5, 5, 5 };
read2.BamAlignment.Qualities = new byte[] { 5, 5, 5, 5, 5, 20, 19, 18 };
alignmentSet = new AlignmentSet(read1, read2);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(read1.Sequence.Substring(3, 5), mergedRead.Sequence.Substring(overlapStart, 5));
Assert.Equal("TTTTTTTTAAA", mergedRead.Sequence);
StitcherTestHelpers.CompareQuality(new byte[] { 30, 30, 30, 5, 5, 5, 5, 5, 20, 19, 18 }, mergedRead.Qualities);
}
public void TryStitch_WithXCTag()
{
const string xcTagDiffFromCalculated = "4M2I4M";
const string expectedCalculatedCigar = "10M";
var read1 = TestHelper.CreateRead("chr1", "ATCGATCG", 12345,
new CigarAlignment("8M"), qualityForAll: 30);
var read2_overlap = TestHelper.CreateRead("chr1", "ATCGTT", 12349,
new CigarAlignment("6M"), qualityForAll: 30);
var stitcher = StitcherTestHelpers.GetStitcher(10);
// -----------------------------------------------
// XC tag is available, and matching between R1 and R2, and expected length,
// but is different from the cigar string we would have calculated
// -----------------------------------------------
// XC tag should be taken
read1.StitchedCigar = new CigarAlignment(xcTagDiffFromCalculated);
read2_overlap.StitchedCigar = new CigarAlignment(xcTagDiffFromCalculated);
var alignmentSet = new AlignmentSet(read1, read2_overlap);
stitcher.TryStitch(alignmentSet);
var mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(xcTagDiffFromCalculated, mergedRead.CigarData.ToString());
// -----------------------------------------------
// XC tag is there, and matching between R1 and R2, but not expected length
// -----------------------------------------------
// XC tag should be ignored if it is not expected length, and new cigar should be calculated
read1.StitchedCigar = new CigarAlignment("8M");
read2_overlap.StitchedCigar = new CigarAlignment("8M");
alignmentSet = new AlignmentSet(read1, read2_overlap);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(expectedCalculatedCigar, mergedRead.CigarData.ToString());
// -----------------------------------------------
// XC tag is there on one read but not the other
// -----------------------------------------------
// XC tag should be ignored, and new cigar should be calculated
read1.StitchedCigar = null;
read2_overlap.StitchedCigar = new CigarAlignment("9M1I");
alignmentSet = new AlignmentSet(read1, read2_overlap);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(expectedCalculatedCigar, mergedRead.CigarData.ToString());
read1.StitchedCigar = new CigarAlignment("9M1I");
read2_overlap.StitchedCigar = null;
alignmentSet = new AlignmentSet(read1, read2_overlap);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(expectedCalculatedCigar, mergedRead.CigarData.ToString());
// -----------------------------------------------
// XC tag is not there
// -----------------------------------------------
// New cigar should be calculated
read1.StitchedCigar = null;
read2_overlap.StitchedCigar = null;
alignmentSet = new AlignmentSet(read1, read2_overlap);
stitcher.TryStitch(alignmentSet);
mergedRead = StitcherTestHelpers.GetMergedRead(alignmentSet);
Assert.Equal(expectedCalculatedCigar, mergedRead.CigarData.ToString());
}
public void TryStitch_WithXCTag()
{
const string xcTagDiffFromCalculated = "4M2I4M";
const string expectedCalculatedCigar = "10M";
var xcRequiredStitcher = GetXCStitcher();
// -----------------------------------------------
// XC tag is available, and matching between R1 and R2, and expected length,
// but is different from the cigar string we would have calculated
// -----------------------------------------------
// [If require XC]
// XC tag should be taken
var alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
alignmentSet.PartnerRead1.StitchedCigar = new CigarAlignment(xcTagDiffFromCalculated);
alignmentSet.PartnerRead2.StitchedCigar = new CigarAlignment(xcTagDiffFromCalculated);
xcRequiredStitcher.TryStitch(alignmentSet);
Assert.Equal(1, alignmentSet.ReadsForProcessing.Count);
var mergedRead = alignmentSet.ReadsForProcessing.First();
Assert.Equal(xcTagDiffFromCalculated, mergedRead.CigarData.ToString());
// -----------------------------------------------
// XC tag is there, and matching between R1 and R2, but not expected length
// -----------------------------------------------
// [If not require XC]
//bounce back to processing separately?
//alignmentSet = GetOverlappingReadSet();
//alignmentSet.PartnerRead1.StitchedCigarString = "8M";
//alignmentSet.PartnerRead2.StitchedCigarString = "8M";
//stitcher.TryStitch(alignmentSet);
//Assert.Equal(2, alignmentSet.ReadsForProcessing.Count);
// [If require XC]
//???
// -----------------------------------------------
// XC tag is there on one read but not the other
// -----------------------------------------------
// [If require XC]
// should bounce back to separate processing
alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
alignmentSet.PartnerRead1.StitchedCigar = new CigarAlignment("4M2I4M");
alignmentSet.PartnerRead2.StitchedCigar = null;
StitcherTestHelpers.TryStitchAndAssertFailed(xcRequiredStitcher, alignmentSet);
alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
alignmentSet.PartnerRead1.StitchedCigar = null;
alignmentSet.PartnerRead2.StitchedCigar = new CigarAlignment("4M2I4M");
StitcherTestHelpers.TryStitchAndAssertFailed(xcRequiredStitcher, alignmentSet);
// -----------------------------------------------
// XC tag is not there
// -----------------------------------------------
// [If require XC]
// should bounce back to separate processing
alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
alignmentSet.PartnerRead1.StitchedCigar = null;
alignmentSet.PartnerRead2.StitchedCigar = null;
StitcherTestHelpers.TryStitchAndAssertFailed(xcRequiredStitcher, alignmentSet);
// -----------------------------------------------
// XC tag does not match between read1 and read2
// -----------------------------------------------
// [If require XC]
// should bounce back to separate processing
alignmentSet = StitcherTestHelpers.GetOverlappingReadSet();
alignmentSet.PartnerRead1.StitchedCigar = new CigarAlignment("4M2I4M");
alignmentSet.PartnerRead2.StitchedCigar = new CigarAlignment("9M1I");
StitcherTestHelpers.TryStitchAndAssertFailed(xcRequiredStitcher, alignmentSet);
}
private IAlignmentStitcher GetXCStitcher()
{
return(StitcherTestHelpers.GetStitcher(10, true));
}
public void CallSomaticVariants_MergeReadsWithInsertion_BoundaryTests()
{
//Migrated from old Pisces: Originally called CallSomaticVariants_MergeReadsWithInsertion_BoundaryTests
// insertion at edge of read
//0 1 2 3 - - - 4 5 6 7 8 9
//- C A T A T A G G
//- - - - A T A G G T A A
var read1 = TestHelper.CreateRead("chr1",
"CATATAGG",
1,
new CigarAlignment("3M3I2M"),
new byte[8],
4);
var read2 = TestHelper.CreateRead("chr1",
"ATAGGTAA",
4,
new CigarAlignment("3S5M"),
new byte[8],
1);
TestSuccesfullyStitchedRead(read1, read2, 0, "3M3I5M", (mergedRead) =>
{
Assert.NotNull(mergedRead);
Assert.Equal(mergedRead.Sequence, "CATATAGGTAA");
});
//0 1 2 3 - - - 4 5 6 7 8 9
//- C A T A T A G G
//- - - - - T A G G T A A
read1 = TestHelper.CreateRead("chr1", "CATATAGG", 1, new CigarAlignment("3M3I2M"),
new byte[8], 4);
read2 = TestHelper.CreateRead("chr1", "TAGGTAA", 4, new CigarAlignment("2S5M"),
new byte[8], 1);
TestSuccesfullyStitchedRead(read1, read2, 0, "3M3I5M", (mergedRead) =>
{
Assert.NotNull(mergedRead);
Assert.Equal(mergedRead.Sequence, "CATATAGGTAA");
});
// shouldnt be able to stitch soft clipped areas only if XC tag not provided
//0 1 2 3 - - - 4 5 6 7 8 9
//- C A T A T A G G
//- - - - - T A G G T A A
read1 = TestHelper.CreateRead("chr1", "CATATAGG", 1, new CigarAlignment("3M5S"),
new byte[8], 4);
read2 = TestHelper.CreateRead("chr1", "TAGGTAA", 4, new CigarAlignment("2S5M"),
new byte[8], 1);
StitcherTestHelpers.TestUnstitchableReads(read1, read2, 0, (unStitchableReads) =>
{
Assert.Equal(1, unStitchableReads.Count(x => StitcherTestHelpers.VerifyReadsEqual(read1, x)));
Assert.Equal(1, unStitchableReads.Count(x => StitcherTestHelpers.VerifyReadsEqual(read2, x)));
});
}
public void TryStitch_MergeReadsSmall()
{
//Migrated from old Pisces: Originally called CallSomaticVariants_MergeReadsSmall
//test1: happy path
//0 1 2 3 4 5 6 7 8 9
//- C A T A T
//- - - - A T A G G
var read1 = TestHelper.CreateRead("chr1", "CATAT", 1, new CigarAlignment("5M"), new byte[] { 1, 2, 3, 4, 5 }, 4);
StitcherTestHelpers.SetReadDirections(read1, DirectionType.Forward);
var read2 = TestHelper.CreateRead("chr1", "ATAGG", 4, new CigarAlignment("5M"), new byte[] { 1, 20, 30, 40, 50 }, 1);
StitcherTestHelpers.SetReadDirections(read2, DirectionType.Reverse);
var alignmentSet = new AlignmentSet(read1, read2);
var stitcher = StitcherTestHelpers.GetStitcher(10);
stitcher.TryStitch(alignmentSet);
TestSuccesfullyStitchedRead(read1, read2, 0, "8M", (mergedRead) =>
{
Assert.Equal(mergedRead.Sequence, "CATATAGG");
StitcherTestHelpers.CompareQuality(new byte[] { 1, 2, 3, 4, 20, 30, 40, 50 }, mergedRead.Qualities);
var expectedDirections = StitcherTestHelpers.BuildDirectionMap(new List <IEnumerable <DirectionType> >
{
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Forward, 3),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Stitched, 2),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Reverse, 3)
});
StitcherTestHelpers.VerifyDirectionType(expectedDirections, mergedRead.DirectionMap);
});
//test2: different bases, one with low Q
//0 1 2 3 4 5 6 7 8 9
//- C A T A G
//- - - - A T A G G
read1 = TestHelper.CreateRead("chr1", "CATAG", 1, new CigarAlignment("5M"), new byte[] { 1, 2, 3, 4, 5 }, 4);
StitcherTestHelpers.SetReadDirections(read1, DirectionType.Reverse);
read2 = TestHelper.CreateRead("chr1", "ATAGG", 4, new CigarAlignment("5M"), new byte[] { 1, 20, 30, 40, 50 },
1);
StitcherTestHelpers.SetReadDirections(read2, DirectionType.Forward);
TestSuccesfullyStitchedRead(read1, read2, 10, "8M", (mergedRead) =>
{
Assert.NotNull(mergedRead);
Assert.Equal(mergedRead.Sequence, "CATATAGG");
StitcherTestHelpers.CompareQuality(new byte[] { 1, 2, 3, 4, 20, 30, 40, 50 }, mergedRead.Qualities);
var expectedDirections = StitcherTestHelpers.BuildDirectionMap(new List <IEnumerable <DirectionType> >
{
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Reverse, 3),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Stitched, 2),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Forward, 3)
});
StitcherTestHelpers.VerifyDirectionType(expectedDirections, mergedRead.DirectionMap);
});
//test3: different bases, both with high Q
//0 1 2 3 4 5 6 7 8 9
//- C A T A G
//- - - - A T A G G
read1 = TestHelper.CreateRead("chr1", "CATAG", 1, new CigarAlignment("5M"), new byte[] { 100, 200, 200, 200, 200 }, 4);
read2 = TestHelper.CreateRead("chr1", "ATAGG", 4, new CigarAlignment("5M"), new byte[] { 1, 20, 30, 40, 50 }, 1);
StitcherTestHelpers.SetReadDirections(read1, DirectionType.Forward);
StitcherTestHelpers.SetReadDirections(read2, DirectionType.Reverse);
TestSuccesfullyStitchedRead(read1, read2, 10, "8M", (mergedRead) =>
{
Assert.NotNull(mergedRead);
Assert.Equal(mergedRead.Sequence, "CATANAGG");
StitcherTestHelpers.CompareQuality(new byte[] { 100, 200, 200, 200, 0, 30, 40, 50 }, mergedRead.Qualities);
var expectedDirections = StitcherTestHelpers.BuildDirectionMap(new List <IEnumerable <DirectionType> >
{
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Forward, 3),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Stitched, 2),
StitcherTestHelpers.BuildDirectionSegment(DirectionType.Reverse, 3)
});
StitcherTestHelpers.VerifyDirectionType(expectedDirections, mergedRead.DirectionMap);
});
}