public void Expander()
{
var directionString = "2F3S2R";
var directions = new CigarDirection(directionString);
var expandedDirections = directions.Expand();
int i = 0;
for (var expander = new CigarDirectionExpander(directions); expander.IsNotEnd(); expander.MoveNext())
{
Assert.Equal(expandedDirections[i], expander.Current);
++i;
}
}
public Read GenerateConsensusReadForSimple(Read read1, Read read2, StitchingInfo stitchingInfo, bool isOutie)
{
_stitchedBases.Clear();
_stitchedQualities.Clear();
var startIndexInR1 = 0;
var startIndexInR2 = 0;
var r1PrefixClip = (int)read1.CigarData.GetPrefixClip();
var r2PrefixClip = (int)read2.CigarData.GetPrefixClip();
var r1SuffixClipEnd = (int)read1.CigarData.GetReadSpan();
var r2SuffixClipEnd = (int)read2.CigarData.GetReadSpan();
var r1SuffixClipBegin = r1SuffixClipEnd - read1.CigarData.GetSuffixClip();
var r2SuffixClipBegin = r2SuffixClipEnd - read2.CigarData.GetSuffixClip();
if (!_useSoftclippedBases)
{
if (r2PrefixClip == 0)
{
// If we're not using softclipped bases to count toward stitching-ness, we need to fast-forward ahead of the sofctclip bases in R2
// That way, we essentially ignore the R2 softclipped bases and start from the "real" calls once we get to the Reverse-only region
startIndexInR2 += (int)read2.CigarData.GetPrefixClip();
}
}
if (_ignoreProbeSoftclips)
{
if (isOutie)
{
startIndexInR2 += stitchingInfo.IgnoredProbePrefixBases;
}
else if (r2PrefixClip == 0)
{
startIndexInR1 += stitchingInfo.IgnoredProbePrefixBases;
}
}
var r1Indexer = new ReadIndexer(startIndexInR1);
var r2Indexer = new ReadIndexer(startIndexInR2);
ReadIndexer forwardReadIndexer;
ReadIndexer reverseReadIndexer;
var read1Reverse = false;
// Assumption is that exactly one read is forward and one read is reverse, and each component read is only one direction
if (read1.SequencedBaseDirectionMap.First() == DirectionType.Forward)
{
forwardReadIndexer = r1Indexer;
reverseReadIndexer = r2Indexer;
}
else
{
read1Reverse = true;
forwardReadIndexer = r2Indexer;
reverseReadIndexer = r1Indexer;
}
var r1SoftclipBeforeR2 = read2.ClipAdjustedPosition - read1.ClipAdjustedPosition;
var r2SoftclipBeforeR1 = read1.ClipAdjustedPosition - read2.ClipAdjustedPosition;
CigarDirectionExpander cigarDirectionExpander = new CigarDirectionExpander(stitchingInfo.StitchedDirections);
for (CigarExtensions.CigarOpExpander cigarExpander = new CigarExtensions.CigarOpExpander(stitchingInfo.StitchedCigar);
cigarExpander.IsNotEnd() && cigarDirectionExpander.IsNotEnd();
cigarExpander.MoveNext(), cigarDirectionExpander.MoveNext())
{
var cigarType = cigarExpander.Current;
var direction = cigarDirectionExpander.Current;
if (cigarType == 'D')
{
continue;
}
var r1Index = r1Indexer.Index;
var r2Index = r2Indexer.Index;
if (r1SoftclipBeforeR2 > 0)
{
if (r1Index == r1SoftclipBeforeR2 && !r2Indexer.StartedIndexing)
{
r2Indexer.StartIndexing();
}
}
else if (r2SoftclipBeforeR1 > 0)
{
if (r2Index == r2SoftclipBeforeR1 && !r1Indexer.StartedIndexing)
{
r1Indexer.StartIndexing();
}
}
else
{
r1Indexer.StartIndexing();
r2Indexer.StartIndexing();
}
// Start moving in read if needed
switch (direction)
{
case DirectionType.Forward:
if (!forwardReadIndexer.StartedIndexing)
{
forwardReadIndexer.StartIndexing();
}
break;
case DirectionType.Reverse:
if (!reverseReadIndexer.StartedIndexing)
{
reverseReadIndexer.StartIndexing();
}
break;
case DirectionType.Stitched:
if (!forwardReadIndexer.StartedIndexing)
{
forwardReadIndexer.StartIndexing();
}
if (!reverseReadIndexer.StartedIndexing)
{
reverseReadIndexer.StartIndexing();
}
break;
}
var forwardIndex = read1Reverse ? r2Index : r1Index;
var reverseIndex = read1Reverse ? r1Index : r2Index;
var forwardPrefixClip = read1Reverse ? r2PrefixClip : r1PrefixClip;
var reversePrefixClip = read1Reverse ? r1PrefixClip : r2PrefixClip;
var reverseSuffixClipEnd = read1Reverse ? r1SuffixClipEnd : r2SuffixClipEnd;
var forwardSuffixClipEnd = read1Reverse ? r2SuffixClipEnd : r1SuffixClipEnd;
var forwardSuffixClipBegin = read1Reverse ? r2SuffixClipBegin : r1SuffixClipBegin;
var reverseSuffixClipBegin = read1Reverse ? r1SuffixClipBegin : r2SuffixClipBegin;
// If R1 & R2 are both in prefix softclips, favor R2 as more "real" and skip over the R1 base
if (forwardReadIndexer.StartedIndexing && forwardIndex >= 0 && forwardIndex < forwardPrefixClip)
{
if (reverseReadIndexer.StartedIndexing && reverseIndex >= 0 && reverseIndex < reversePrefixClip)
{
direction = DirectionType.Reverse;
forwardReadIndexer.Increment();
}
}
// If R1 & R2 are both in suffix softclips, favor R1 as more "real" and skip over the R2 base
if (reverseReadIndexer.StartedIndexing && reverseIndex >= reverseSuffixClipBegin && reverseIndex < reverseSuffixClipEnd)
{
if (forwardReadIndexer.StartedIndexing && forwardIndex >= forwardSuffixClipBegin && forwardIndex < forwardSuffixClipEnd)
{
direction = DirectionType.Forward;
reverseReadIndexer.Increment();
}
}
if (r1Index >= 0 && r1Index < read1.BamAlignment.Bases.Length)
{
r1Indexer.BaseAtIndex = read1.BamAlignment.Bases[r1Index];
r1Indexer.QualityAtIndex = read1.Qualities[r1Index];
}
else
{
r1Indexer.BaseAtIndex = null;
r1Indexer.QualityAtIndex = null;
}
if (r2Index >= 0 && r2Index < read2.BamAlignment.Bases.Length)
{
r2Indexer.BaseAtIndex = read2.BamAlignment.Bases[r2Index];
r2Indexer.QualityAtIndex = read2.Qualities[r2Index];
}
else
{
r2Indexer.BaseAtIndex = null;
r2Indexer.QualityAtIndex = null;
}
switch (direction)
{
case DirectionType.Forward:
if (forwardReadIndexer.BaseAtIndex == null)
{
throw new InvalidDataException("Forward base at index " + forwardReadIndexer.Index + " is null.");
}
_stitchedBases.Add((char)forwardReadIndexer.BaseAtIndex);
_stitchedQualities.Add((byte)forwardReadIndexer.QualityAtIndex);
forwardReadIndexer.Increment();
break;
case DirectionType.Reverse:
if (reverseReadIndexer.BaseAtIndex == null)
{
throw new InvalidDataException("Reverse base at index " + reverseReadIndexer.Index + " is null.");
}
_stitchedBases.Add((char)reverseReadIndexer.BaseAtIndex); // TODO - stringbuilder instead?
_stitchedQualities.Add((byte)reverseReadIndexer.QualityAtIndex);
reverseReadIndexer.Increment();
break;
case DirectionType.Stitched:
if (forwardReadIndexer.BaseAtIndex != null && reverseReadIndexer.BaseAtIndex != null)
{
if (forwardReadIndexer.BaseAtIndex == reverseReadIndexer.BaseAtIndex)
{
_stitchedBases.Add((char)forwardReadIndexer.BaseAtIndex);
var sumQuality = Convert.ToInt32((byte)forwardReadIndexer.QualityAtIndex) +
Convert.ToInt32((byte)reverseReadIndexer.QualityAtIndex);
var sticheredQuality = sumQuality > MaxBaseQuality ? MaxBaseQuality : sumQuality;
_stitchedQualities.Add((byte)sticheredQuality);
}
else //the bases disagree...
{
if (_nifyDisagreements)
{
// we have disagreeing bases AND we chose to always Nify them
_stitchedBases.Add('N');
_stitchedQualities.Add(0);
}
else
{
if ((byte)forwardReadIndexer.QualityAtIndex >= reverseReadIndexer.QualityAtIndex)
// Original stitching implementation -- TODO, reconcile this with new reqs.
{
_stitchedBases.Add((char)forwardReadIndexer.BaseAtIndex);
if (reverseReadIndexer.QualityAtIndex < _minBasecallQuality)
{
_stitchedQualities.Add((byte)forwardReadIndexer.QualityAtIndex);
}
else
{
_stitchedQualities.Add(0);
}
//this was a high Q disagreement, and dangerous! we will filter this base.
}
else
//if ((byte)forwardReadIndexer.QualityAtIndex < reverseReadIndexer.QualityAtIndex) // Original stitching implementation
{
_stitchedBases.Add((char)reverseReadIndexer.BaseAtIndex);
if (forwardReadIndexer.QualityAtIndex < _minBasecallQuality)
{
_stitchedQualities.Add((byte)reverseReadIndexer.QualityAtIndex);
}
else
{
_stitchedQualities.Add(0);
}
//this was a high Q disagreement, and dangerous! we will filter this base.
}
}
}
}
forwardReadIndexer.Increment();
reverseReadIndexer.Increment();
break;
default:
throw new ArgumentOutOfRangeException();
}
}
// Validate stitched cigar
var r2CigarLength = read2.CigarData.Cast <CigarOp>().Sum(op => (int)op.Length);
var r1CigarLength = read1.CigarData.Cast <CigarOp>().Sum(op => (int)op.Length);
var stitchedCigarLength = stitchingInfo.StitchedCigar.Cast <CigarOp>().Sum(op => (int)op.Length);
var earliestStart = Math.Min(read1.ClipAdjustedPosition, read2.ClipAdjustedPosition);
var latestEnd = Math.Max(read1.ClipAdjustedPosition + r1CigarLength, read2.ClipAdjustedPosition + r2CigarLength);
//var latestEnd = Math.Max(read1.ClipAdjustedPosition + read1.CigarData.GetReadSpan(), read2.ClipAdjustedPosition + read2.CigarData+ stitchingInfo.InsertionAdjustment);
if (stitchedCigarLength != (latestEnd - earliestStart))
{
// TODO what is really the point of this???
if (_debug)
{
Logger.WriteToLog(string.Format(
"Attempted stitched cigar {0} is not consistent with component reads {1}:{2} and {3}:{4}",
stitchingInfo.StitchedCigar, read1.Position, read1.CigarData, read2.Position,
read2.CigarData));
}
_statusCounter.AddDebugStatusCount("Attempted stitched cigar not consistent with component reads");
//return null;
}
// TODO investigate if these are ever worth handling
if (stitchingInfo.StitchedCigar.Count > 0 && stitchingInfo.StitchedCigar.GetReadSpan() != _stitchedBases.Count)
{
if (_debug)
{
Logger.WriteToLog(string.Format("Invalid cigar '{0}': does not match length {1} of read ({2})", stitchingInfo.StitchedCigar,
_stitchedBases.Count, read1.Name));
}
_statusCounter.AddDebugStatusCount("Invalid cigar does not match length of read");
return(null);
}
var mergedRead = new Read(read1.Chromosome, new BamAlignment
{
Name = read1.Name,
Bases = string.Join("", _stitchedBases),
Position = Math.Min(read1.Position - 1, read2.Position - 1),
Qualities = _stitchedQualities.ToArray(),
CigarData = stitchingInfo.StitchedCigar
})
{
StitchedCigar = stitchingInfo.StitchedCigar
};
return(mergedRead);
}