private static BiasResults GetProbePoolBiasScore(VariantComparisonCase Case, CalledAllele Consensus,
float ProbePoolBiasThreshold, VariantCallingParameters variantCallingParameters, int AltCountA, int AltCountB, int DepthA, int DepthB, Genotype Genotype, bool AltChangeToRef)
{
double ProbePoolPScore = 0; //no bias;
double ProbePoolGATKBiasScore = -100; //no bias;
int NoiseLevel = Consensus.NoiseLevelApplied;
BiasResults PB = new BiasResults();
if ((AltChangeToRef) || (Case == VariantComparisonCase.AgreedOnReference))
{
PB.GATKBiasScore = ProbePoolGATKBiasScore;
PB.BiasScore = ProbePoolPScore;
return(PB);
}
if ((Case == VariantComparisonCase.OneReferenceOneAlternate) ||
(Case == VariantComparisonCase.CanNotCombine))
{
Consensus.Filters.Add(FilterType.PoolBias);
PB.GATKBiasScore = 0;
PB.BiasScore = 1;
return(PB);
}
if (Case == VariantComparisonCase.AgreedOnAlternate)
{
int[] supportByPool = new int[]
{
AltCountA, AltCountB, 0
};
int[] covByPool = new int[]
{
DepthA, DepthB, 0
};
BiasResults ProbePoolBiasResults =
StrandBiasCalculator.CalculateStrandBiasResults(
covByPool, supportByPool,
NoiseLevel, variantCallingParameters.MinimumFrequency, ProbePoolBiasThreshold, StrandBiasModel.Extended);
ProbePoolGATKBiasScore = Math.Min(0, ProbePoolBiasResults.GATKBiasScore); //just cap it at upperbound 0, dont go higher.
ProbePoolGATKBiasScore = Math.Max(-100, ProbePoolGATKBiasScore); //just cap it at lowerbound -100, dont go higher.
ProbePoolPScore = Math.Min(1, ProbePoolBiasResults.BiasScore);
if (!ProbePoolBiasResults.BiasAcceptable)
{
Consensus.Filters.Add(FilterType.PoolBias);
}
}
PB.GATKBiasScore = ProbePoolGATKBiasScore;
PB.BiasScore = ProbePoolPScore;
return(PB);
}
public void TestSBCalculationsForForcedVariants()
{
var CoverageByStrandDirection = new int[] { 70038, 65998, 0 }; //forward,reverse,stitched
var SupportByStrandDirection = new int[] { 54, 11, 0 };
BiasResults SB = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, SupportByStrandDirection, 20, 0.01, 0.5, StrandBiasModel.Poisson);
Assert.Equal(SB.BiasScore, 1.0);
Assert.Equal(SB.GATKBiasScore, 0);
}
public CalledAllele()
{
Filters = new List <FilterType>();
EstimatedCoverageByDirection = new int[Constants.NumDirectionTypes];
StrandBiasResults = new BiasResults();
SupportByDirection = new int[Constants.NumDirectionTypes];
WellAnchoredSupportByDirection = new int[Constants.NumDirectionTypes];
ReadCollapsedCountsMut = new int[Constants.NumReadCollapsedTypes];
ReadCollapsedCountTotal = new int[Constants.NumReadCollapsedTypes];
Genotype = Genotype.HomozygousRef;
Type = AlleleCategory.Reference;
}
/// <summary>
/// Assign a strandbias-score to a SNP.
/// (using only forward and reverse SNP counts.)
/// </summary>
public static BiasResults CalculateStrandBiasResults(int[] coverageByStrandDirection,
int[] supportByStrandDirection,
int qNoise, double minVariantFreq, double acceptanceCriteria, StrandBiasModel strandBiasModel)
{
var forwardSupport = supportByStrandDirection[(int)DirectionType.Forward];
var forwardCoverage = coverageByStrandDirection[(int)DirectionType.Forward];
var reverseSupport = supportByStrandDirection[(int)DirectionType.Reverse];
var reverseCoverage = coverageByStrandDirection[(int)DirectionType.Reverse];
var stitchedSupport = supportByStrandDirection[(int)DirectionType.Stitched];
var stitchedCoverage = coverageByStrandDirection[(int)DirectionType.Stitched];
var errorRate = Math.Pow(10, -1 * qNoise / 10f);
var overallStats = CreateStats(forwardSupport + reverseSupport + stitchedSupport,
forwardCoverage + reverseCoverage + stitchedCoverage, errorRate, minVariantFreq, strandBiasModel);
var forwardStats = CreateStats(forwardSupport + stitchedSupport / 2,
forwardCoverage + stitchedCoverage / 2,
errorRate, minVariantFreq, strandBiasModel);
var reverseStats = CreateStats(reverseSupport + stitchedSupport / 2,
reverseCoverage + stitchedCoverage / 2,
errorRate, minVariantFreq, strandBiasModel);
var results = new BiasResults
{
ForwardStats = forwardStats,
ReverseStats = reverseStats,
OverallStats = overallStats
};
results.StitchedStats = CreateStats(stitchedSupport, stitchedCoverage, errorRate, minVariantFreq,
strandBiasModel);
var biasResults = AssignBiasScore(overallStats, forwardStats, reverseStats);
results.BiasScore = biasResults[0];
results.GATKBiasScore = biasResults[1];
results.CovPresentOnBothStrands = ((forwardStats.Coverage > 0) && (reverseStats.Coverage > 0));
results.VarPresentOnBothStrands = ((forwardStats.Support > 0) && (reverseStats.Support > 0));
//not really fair to call it biased if coverage is in one direction..
//its ambiguous if variant is found in only one direction.
if (!results.CovPresentOnBothStrands)
{
results.BiasScore = 0;
results.GATKBiasScore = double.NegativeInfinity;
}
results.BiasAcceptable = (results.BiasScore < acceptanceCriteria);
return(results);
}
public CalledAllele(CalledAllele originalAllele)
{
Chromosome = originalAllele.Chromosome;
ReferencePosition = originalAllele.ReferencePosition;
ReferenceAllele = originalAllele.ReferenceAllele;
AlternateAllele = originalAllele.AlternateAllele;
Genotype = originalAllele.Genotype;
GenotypeQscore = originalAllele.GenotypeQscore;
VariantQscore = originalAllele.VariantQscore;
NumNoCalls = originalAllele.NumNoCalls;
NoiseLevelApplied = originalAllele.NoiseLevelApplied;
TotalCoverage = originalAllele.TotalCoverage;
AlleleSupport = originalAllele.AlleleSupport;
WellAnchoredSupport = originalAllele.WellAnchoredSupport;
ReferenceSupport = originalAllele.ReferenceSupport;
Type = originalAllele.Type;
SupportByDirection = new int[Constants.NumDirectionTypes];
WellAnchoredSupportByDirection = new int[Constants.NumDirectionTypes];
EstimatedCoverageByDirection = new int[Constants.NumDirectionTypes];
ReadCollapsedCountsMut = new int[Constants.NumReadCollapsedTypes];
ReadCollapsedCountTotal = new int[Constants.NumReadCollapsedTypes];
StrandBiasResults = BiasResults.DeepCopy(originalAllele.StrandBiasResults);
UnanchoredCoverageWeight = originalAllele.UnanchoredCoverageWeight;
ConfidentCoverageStart = originalAllele.ConfidentCoverageStart;
ConfidentCoverageEnd = originalAllele.ConfidentCoverageEnd;
SuspiciousCoverageStart = originalAllele.SuspiciousCoverageStart;
SuspiciousCoverageEnd = originalAllele.SuspiciousCoverageEnd;
Filters = new List <FilterType>();
foreach (var filter in originalAllele.Filters)
{
Filters.Add(filter);
}
for (int i = 0; i < Constants.NumDirectionTypes; i++)
{
SupportByDirection[i] = originalAllele.SupportByDirection[i];
WellAnchoredSupportByDirection[i] = originalAllele.WellAnchoredSupportByDirection[i];
EstimatedCoverageByDirection[i] = originalAllele.EstimatedCoverageByDirection[i];
}
for (int i = 0; i < Constants.NumReadCollapsedTypes; i++)
{
ReadCollapsedCountsMut[i] = originalAllele.ReadCollapsedCountsMut[i];
ReadCollapsedCountTotal[i] = originalAllele.ReadCollapsedCountTotal[i];
}
}
private static BiasResults GetCombinedSBValue(CalledAllele VariantA, CalledAllele VariantB, SampleAggregationParameters SampleAggregationOptions)
{
BiasResults StrandBiasResults = new BiasResults();
if (VariantA == null)
{
return(VariantB.StrandBiasResults);
}
if (VariantB == null)
{
return(VariantA.StrandBiasResults);
}
StrandBiasResults.GATKBiasScore = Math.Max(VariantA.StrandBiasResults.GATKBiasScore, VariantB.StrandBiasResults.GATKBiasScore);
return(StrandBiasResults);
}
public static string StatsToString(BiasResults stat)
{
var delimiter = "\t";
StringBuilder builder = new StringBuilder();
string[] statsData = new string[3 * 5];
StringHelper(stat.OverallStats, statsData, 0, 3, delimiter);
StringHelper(stat.ForwardStats, statsData, 1, 3, delimiter);
StringHelper(stat.ReverseStats, statsData, 2, 3, delimiter);
foreach (string t in statsData)
{
builder.Append(t);
}
//raw data
//for (int i = 0; i < Constants.NumDirectionTypes; i++)
// builder.Append(stat.CoverageByStrandDirection[i] + "\t");
builder.Append(stat.ForwardStats.Coverage + delimiter);
builder.Append(stat.ReverseStats.Coverage + delimiter);
builder.Append(stat.StitchedStats.Coverage + delimiter);
//for (int i = 0; i < Constants.NumDirectionTypes; i++)
// builder.Append(SupportByStrandDirection[i] + "\t");
builder.Append(stat.ForwardStats.Support + delimiter);
builder.Append(stat.ReverseStats.Support + delimiter);
builder.Append(stat.StitchedStats.Support + delimiter);
//results
builder.Append(stat.BiasScore + delimiter);
builder.Append(stat.BiasAcceptable + delimiter);
builder.Append(stat.VarPresentOnBothStrands + delimiter);
builder.Append(stat.CovPresentOnBothStrands + delimiter);
return(builder.ToString());
}
public static BiasResults DeepCopy(BiasResults originalSBresults)
{
if (originalSBresults == null)
{
return(null);
}
var sb = new BiasResults()
{
BiasAcceptable = originalSBresults.BiasAcceptable,
BiasScore = originalSBresults.BiasScore,
GATKBiasScore = originalSBresults.GATKBiasScore,
VarPresentOnBothStrands = originalSBresults.VarPresentOnBothStrands,
CovPresentOnBothStrands = originalSBresults.CovPresentOnBothStrands,
TestAcceptable = originalSBresults.TestAcceptable,
TestScore = originalSBresults.TestScore,
ForwardStats = StrandBiasStats.DeepCopy(originalSBresults.ForwardStats),
OverallStats = StrandBiasStats.DeepCopy(originalSBresults.OverallStats),
ReverseStats = StrandBiasStats.DeepCopy(originalSBresults.ReverseStats),
StitchedStats = StrandBiasStats.DeepCopy(originalSBresults.StitchedStats),
};
return(sb);
}
public void TestSBCalculationsForSomaticAndDiploidSettings()
{
double fwdCov = 10000;
double revCov = 10000;
double testVariantFreqA = 0.05;
double testVariantFreqB = 0.25;
double testVariantFreqC = 0.020;
double testVariantFreqD = 0.005;
var CoverageByStrandDirection = new int[] { (int)fwdCov, (int)revCov, 0 }; //forward,reverse,stitched
var EqualSupportByStrandDirectionA = new int[] { (int)(fwdCov * testVariantFreqA), (int)(revCov * testVariantFreqA), 0 };
var EqualSupportByStrandDirectionB = new int[] { (int)(fwdCov * testVariantFreqB), (int)(revCov * testVariantFreqB), 0 };
//happy path, no bias
BiasResults SB_somatic = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, EqualSupportByStrandDirectionB, 20, 0.01, 0.5, StrandBiasModel.Extended);
BiasResults SB_diploid = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, EqualSupportByStrandDirectionB, 20, 0.20, 0.5, StrandBiasModel.Diploid);
Assert.Equal(SB_somatic.BiasScore, 0);
Assert.Equal(SB_somatic.GATKBiasScore, double.NegativeInfinity);
Assert.Equal(SB_somatic.BiasAcceptable, true);
Assert.Equal(SB_diploid.BiasScore, 0);
Assert.Equal(SB_diploid.GATKBiasScore, double.NegativeInfinity);
Assert.Equal(SB_diploid.BiasAcceptable, true);
//bias if you are looking for a 20% variant (only one side is sufficient to call),
//but not biased in the somatic case (both show up sufficiently)
var SupportByStrandDirection_bias20 = new int[] { (int)(fwdCov * testVariantFreqA), (int)(revCov * testVariantFreqB), 0 };
SB_somatic = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, SupportByStrandDirection_bias20, 20, 0.01, 0.5, StrandBiasModel.Extended);
SB_diploid = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, SupportByStrandDirection_bias20, 20, 0.20, 0.5, StrandBiasModel.Diploid);
Assert.Equal(SB_somatic.BiasScore, 0);
Assert.Equal(SB_somatic.GATKBiasScore, double.NegativeInfinity);
Assert.Equal(SB_somatic.BiasAcceptable, true);
Assert.Equal(Math.Log10(SB_diploid.BiasScore), 74.3, 1); // a great big bias
Assert.Equal(SB_diploid.GATKBiasScore, 743.5, 1);
Assert.Equal(SB_diploid.BiasAcceptable, false);
//bias if you are looking for even a 1% variant or a 20% variant
var SupportByStrandDirection_bias01 = new int[] { (int)(fwdCov * testVariantFreqC), (int)(revCov * testVariantFreqD), 0 };
SB_somatic = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, SupportByStrandDirection_bias01, 20, 0.01, 0.5, StrandBiasModel.Extended);
SB_diploid = StrandBiasCalculator.CalculateStrandBiasResults(
CoverageByStrandDirection, SupportByStrandDirection_bias01, 20, 0.20, 0.5, StrandBiasModel.Diploid);
Assert.Equal(SB_somatic.BiasScore, 1.000, 3);
Assert.Equal(SB_somatic.GATKBiasScore, 0.002, 3);
Assert.Equal(SB_somatic.BiasAcceptable, false);
Assert.Equal(SB_diploid.BiasScore, 1.000, 3);// a great big bias
Assert.Equal(SB_diploid.GATKBiasScore, 0.000, 3);
Assert.Equal(SB_diploid.BiasAcceptable, false);
}
/// <summary>
/// populates a called allele object given an array of vcf columns
/// </summary>
protected static void ConvertColumnsToVariant(bool shouldTrimComplexAlleles, string[] cols, CalledAllele allele, int alleleIndex)
{
bool shouldOutputRcCounts = true;
bool shouldOutputTsCounts = true;
if ((cols == null) || (cols.Length == 0))
{
allele = null;
return;
}
//set defaults.
var genotypeQscore = 0;
var referenceSupport = 0;
var altSupport = 0;
var genotypeString = "";
var totalCoverage = 0;
var variantQuality = 0.0;
var numAlts = 1;
var noiseLevel = 0;
var fractionNocalls = 0f;
var strandBiasInGATKScaleCoords = -100f;
var tsCounts = new List <string>();
//
//read in simple data
allele.Chromosome = cols[VcfCommon.ChromIndex];
allele.ReferencePosition = int.Parse(cols[VcfCommon.PosIndex]);
allele.ReferenceAllele = cols[VcfCommon.RefIndex];
allele.Filters = VcfVariantUtilities.MapFilterString(cols[VcfCommon.FilterIndex]);
bool gotQual = double.TryParse(cols[VcfCommon.QualIndex], out variantQuality); // CFTR uses a ".", which is not actually legal... (actually, vcf 4.1 does allow the missing value "." here. Strelka uses it)
if (gotQual)
{
allele.VariantQscore = (int)variantQuality;
}
// parse the variant alleles
var variantAlleles = cols[VcfCommon.AltIndex].Split(',');
allele.AlternateAllele = variantAlleles[alleleIndex];
var isRef = (allele.AlternateAllele == ".");
if (isRef)
{
numAlts = 0;
}
else
{
numAlts = variantAlleles.Count();
}
// parse the info field data (presume, single sample)
Dictionary <string, string> InfoFields = ParseInfoFields(cols);
// parse the genotype data (presume, single sample)
List <Dictionary <string, string> > Genotypes = ParseGenotypeData(cols);
//get more complex allele data...
if (InfoFields.ContainsKey("DP"))
{
totalCoverage = Int32.Parse(InfoFields["DP"]);
}
if ((Genotypes.Count > 0) && (Genotypes[0] != null))
{
if (Genotypes[0].ContainsKey("GQ"))
{
genotypeQscore = Int32.Parse(Genotypes[0]["GQ"]);
}
else if (Genotypes[0].ContainsKey("GQX"))
{
genotypeQscore = Int32.Parse(Genotypes[0]["GQX"]);
}
if (Genotypes[0].ContainsKey("GT"))
{
genotypeString = Genotypes[0]["GT"];
}
if (Genotypes[0].ContainsKey("NL"))
{
noiseLevel = Int32.Parse(Genotypes[0]["NL"]);
}
if (Genotypes[0].ContainsKey("NC"))
{
fractionNocalls = float.Parse(Genotypes[0]["NC"]);
}
if (Genotypes[0].ContainsKey("SB"))
{
strandBiasInGATKScaleCoords = float.Parse(Genotypes[0]["SB"]);
}
var ADstrings = new string[] { "0", "0" };
if (Genotypes[0].ContainsKey("AD"))
{
ADstrings = Genotypes[0]["AD"].Split(',');
}
referenceSupport = int.Parse(ADstrings[0]);
//by default alt support is 0.
if ((!isRef) && (ADstrings.Length > 1))
{
altSupport = int.Parse(ADstrings[1]);
}
if (shouldOutputRcCounts)
{
if (Genotypes[0].ContainsKey("US"))
{
tsCounts = Genotypes[0]["US"].Split(',').ToList();
}
}
allele.Genotype = VcfVariantUtilities.MapGTString(genotypeString, numAlts);
//note this awkward vcf line (pisces)
//"chr4\t10\t.\tAA\tGA,G\t0\tPASS\tDP=5394\tGT:GQ:AD:DP:VF:NL:SB:NC\t1/2:0:2387,2000:5394:0.8133:23:0.0000:0.0000";
//and this one
//chr2 19946216.ATGTGTG ATG,ATGTG,A 0 PASS metal = platinum; cgi =.; bwa_freebayes = HD:0,LOOHD: 0; bwa_platypus =.; bwa_gatk3 = HD:2,LOOHD: 2; cortex =.; isaac2 = HD:1,LOOHD: 1; dist2closest = 192 GT 1 | 2
if ((numAlts >= 2) && (Genotypes[0].ContainsKey("AD")))
{
if (ADstrings.Count() <= numAlts) //in this case we never expressedly gave the ref support, so we have to derive it.
{
int totalAltCount = 0;
for (int altIndex = 0; altIndex < numAlts; altIndex++)
{
var altSupportAtIndex = int.Parse(ADstrings[altIndex]);
totalAltCount += altSupportAtIndex;
if (altIndex == alleleIndex)
{
altSupport = altSupportAtIndex;
}
}
referenceSupport = Math.Max(0, totalCoverage - totalAltCount);
}
}
}
var strandBiasResults = new BiasResults();
strandBiasResults.GATKBiasScore = strandBiasInGATKScaleCoords;
//set the remaining data
allele.TotalCoverage = totalCoverage;
allele.AlleleSupport = isRef ? referenceSupport : altSupport;
allele.ReferenceSupport = referenceSupport;
allele.GenotypeQscore = genotypeQscore;
allele.NoiseLevelApplied = noiseLevel;
allele.StrandBiasResults = strandBiasResults;
allele.IsForcedToReport = allele.Filters.Contains(FilterType.ForcedReport);
//set the derived values
allele.SetType();
allele.ForceFractionNoCalls(fractionNocalls);
//rescue attempt for complex types, ie ACGT -> ACGTGG.
//Get the simplest form of the allele
if ((allele.Type == AlleleCategory.Unsupported) && shouldTrimComplexAlleles)
{
VcfVariantUtilities.TrimUnsupportedAlleleType(allele);
}
if (tsCounts.Count != 0)
{
VcfVariantUtilities.FillInCollapsedReadsCount(shouldOutputRcCounts, shouldOutputTsCounts, allele, tsCounts);
}
}
public static CalledAllele ConvertUnpackedVariant(VcfVariant v, bool shouldOutputRcCounts = false,
bool shouldOutputTsCounts = false, bool shouldTrimComplexAlleles = true)
{
if (v == null)
{
return(null);
}
if (v.VariantAlleles.Count() > 1)
{
throw new ArgumentException("This method does not handle crushed vcf format. Use Convert(IEnumerable<VcfVariant> vcfVariants)");
}
var genotypeQscore = 0;
var referenceSupport = 0;
var altSupport = 0;
var genotypeString = "";
var totalCoverage = 0;
var isRef = ((v.VariantAlleles.Count() == 1) && v.VariantAlleles[0] == ".");
var variantQuality = v.Quality;
var numAlts = 1;
var noiseLevel = 1;
var fractionNocalls = 0f;
var strandBiasInGATKScaleCoords = -100f;
var tsCounts = new List <string>();
if (v.InfoFields.ContainsKey("DP"))
{
totalCoverage = Int32.Parse(v.InfoFields["DP"]);
}
if (v.Genotypes.Count > 0)
{
if (v.Genotypes[0].ContainsKey("GQ"))
{
genotypeQscore = Int32.Parse(v.Genotypes[0]["GQ"]);
}
else if (v.Genotypes[0].ContainsKey("GQX"))
{
genotypeQscore = Int32.Parse(v.Genotypes[0]["GQX"]);
}
genotypeString = v.Genotypes[0]["GT"];
if (v.Genotypes[0].ContainsKey("NL"))
{
noiseLevel = Int32.Parse(v.Genotypes[0]["NL"]);
}
if (v.Genotypes[0].ContainsKey("NC"))
{
fractionNocalls = float.Parse(v.Genotypes[0]["NC"]);
}
if (v.Genotypes[0].ContainsKey("SB"))
{
strandBiasInGATKScaleCoords = float.Parse(v.Genotypes[0]["SB"]);
}
var ADstring = new string[] { "0", "0" };
if (v.Genotypes[0].ContainsKey("AD"))
{
ADstring = v.Genotypes[0]["AD"].Split(',');
}
var VFstring = new string[] { "0", "0" };
if (v.Genotypes[0].ContainsKey("VF"))
{
VFstring = v.Genotypes[0]["VF"].Split(',');
}
referenceSupport = int.Parse(ADstring[0]);
altSupport = isRef ? 0 : int.Parse(ADstring[1]);
if (shouldOutputRcCounts)
{
if (v.Genotypes[0].ContainsKey("US"))
{
tsCounts = v.Genotypes[0]["US"].Split(',').ToList();
}
}
if (isRef)
{
numAlts = 0;
}
else
{
numAlts = 1; //note this, method should never get a value here >1. these should be UNPACKED variants
}
}
var strandBiasResults = new BiasResults();
strandBiasResults.GATKBiasScore = strandBiasInGATKScaleCoords;
var filters = MapFilterString(v.Filters);
var allele = new CalledAllele()
{
Chromosome = v.ReferenceName,
ReferencePosition = v.ReferencePosition,
ReferenceAllele = v.ReferenceAllele,
AlternateAllele = v.VariantAlleles[0],
TotalCoverage = totalCoverage,
AlleleSupport = isRef ? referenceSupport : altSupport,
ReferenceSupport = referenceSupport,
VariantQscore = (int)variantQuality,
GenotypeQscore = genotypeQscore,
Genotype = MapGTString(genotypeString, numAlts),
Filters = filters,
NoiseLevelApplied = noiseLevel,
StrandBiasResults = strandBiasResults,
IsForcedToReport = filters.Contains(FilterType.ForcedReport)
};
allele.SetType();
allele.ForceFractionNoCalls(fractionNocalls);
//rescue attempt for complex types, ie ACGT -> ACGTGG
if ((allele.Type == AlleleCategory.Unsupported) && shouldTrimComplexAlleles)
{
TrimUnsupportedAlleleType(allele);
}
FillInCollapsedReadsCount(shouldOutputRcCounts, shouldOutputTsCounts, allele, tsCounts);
return(allele);
}