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/*==========================================================================
This program is free software; you can redistribute it and/or
modify it under the terms of the GNU Lesser General Public
License as published by the Free Software Foundation; either
version 3 of the License, or (at your options) any later version.
This program is distributed in the hope that it will be useful,
but WITHOUT ANY WARRANTY; without even the implied warranty of
MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU
Lesser General Public License for more details.
You should have received a copy of the GNU General Public License
along with this program. If not, see <https://www.gnu.org/licenses/>.
==========================================================================*/
// switches on corrected heterozygote variance in allele ratios (higher sensitivity for hets, at the cost of more false positive snp calls)
#define CORRECTED_HET // requires boost library
// for longer reads such as 454 data (will use different scoring scheme)
//#define READS_454
///// switch on extreme debug output
// #define SNPSTORE_DEBUG
//#define SNPSTORE_DEBUG_CANDPOS
#include <seqan/platform.h>
#include <seqan/sequence.h>
#include <seqan/file.h>
#include <seqan/align.h>
#include <seqan/store.h>
#include <seqan/consensus.h>
#include <seqan/stream.h>
#include <seqan/bam_io.h>
#ifdef STDLIB_VS
#define SEQAN_DEFAULT_TMPDIR "C:\\TEMP\\"
#else
#define SEQAN_DEFAULT_TMPDIR "./"
#endif
//#include "../../../apps/rep_sep/utils.h"
//#include "../../../apps/rep_sep/assembly_parser.h"
//#include "../../../apps/rep_sep/column_scanner.h"
//#include "../../../apps/rep_sep/rgraph.h"
//#include "../../../apps/rep_sep/rep_sep.h"
#include "snp_store.h"
#include <iostream>
#include <fstream>
#include <sstream>
#include <map>
#include <seqan/arg_parse.h>
using namespace std;
using namespace seqan2;
// load entire genome into memory
template <typename TGenomeSet, typename TGenomeSetSize, typename TGenomeNames>
bool loadGenomes(TGenomeSet &genomes, StringSet<CharString> &fileNameList, ::std::map<CharString, TGenomeSetSize> &gIdStringToIdNumMap, TGenomeNames & genomeNames)
{
unsigned gSeqNo = 0;
unsigned filecount = 0;
CharString temp;
clear(genomeNames);
while(filecount < length(fileNameList))
{
clear(temp);
SeqFileIn seqFileIn;
if (!open(seqFileIn, toCString(fileNameList[filecount])))
return false;
unsigned seqCount = 0;
Dna5String seq;
for (; !atEnd(seqFileIn); ++seqCount)
{
readRecord(temp, seq, seqFileIn);
// Trim sequence ID and insert into mapping.
cropAfterFirst(temp, NotFunctor<IsGraph>());
gIdStringToIdNumMap.insert(::std::make_pair(temp, length(genomes))); // keeps the whole fasta ID including white spaces
appendValue(genomeNames, temp);
// Register genome sequence.
appendValue(genomes, seq);
}
gSeqNo += seqCount;
++filecount;
}
resize(genomes, gSeqNo);
return (gSeqNo > 0);
}
// transform global cooridnates to coordinates relative to chromosomal segment
template<typename TFragmentStore, typename TContigPos, typename TOptions>
void
transformCoordinates(TFragmentStore &fragmentStore, TContigPos startCoord, TOptions&)
{
typedef typename TFragmentStore::TAlignedReadStore TMatches;
//typedef typename Value<TMatches>::Type TMatch;
typedef typename Iterator<TMatches,Standard>::Type TMatchIt;
TMatchIt mIt = begin(fragmentStore.alignedReadStore,Standard());
TMatchIt mItEnd = end(fragmentStore.alignedReadStore,Standard());
while(mIt != mItEnd)
{
(*mIt).endPos -= startCoord;
(*mIt).beginPos -= startCoord;
++mIt;
}
}
// copy matches relevant for next window
template<typename TFragmentStore, typename TReadCigars, typename TReadCounts, typename TReadClips, typename TSize, typename TContigPos, typename TOptions>
void
copyNextWindowMatchesAndReads(TFragmentStore &fragmentStore,
TReadCounts &readCounts,
TReadCigars &readCigars,
TReadClips &readClips,
TReadCounts &tmpReadCounts,
typename TFragmentStore::TReadSeqStore &tmpReads,
typename TFragmentStore::TReadStore &tmpRs,
typename TFragmentStore::TAlignedReadStore &tmpMatches,
typename TFragmentStore::TAlignQualityStore &tmpQualities,
TReadClips &tmpReadClips,
TReadCigars &tmpReadCigars,
TSize ,
TContigPos currentWindowEnd,
TOptions &options)
{
typedef typename TFragmentStore::TAlignedReadStore TMatches;
typedef typename Value<TMatches>::Type TMatch;
typedef typename Iterator<TMatches,Standard>::Type TMatchIt;
typedef typename Id<TFragmentStore>::Type TId;
//typedef typename Value<TReadClips>::Type TPair;
SEQAN_ASSERT_EQ(length(fragmentStore.readSeqStore),length(fragmentStore.alignQualityStore));
::std::sort(begin(fragmentStore.alignedReadStore, Standard()), end(fragmentStore.alignedReadStore, Standard()), LessGPos<TMatch>());
if(options._debugLevel > 1 )::std::cout << "Copying matches overlapping more than one window ... \n";
TMatchIt mIt = end(fragmentStore.alignedReadStore,Standard());
TMatchIt mItBegin = begin(fragmentStore.alignedReadStore,Standard());
--mIt;
// We will use minCoord/maxCoord to store the temporarily minimal and maximal coordinates in the window.
int minCoord = std::numeric_limits<int>::max();
int maxCoord = std::numeric_limits<int>::min();
//CharString str = "discBef";
//_dumpMatches(fragmentStore, str);
//std::cout << " max hit length = " << options.maxHitLength << std::endl;
// look for matches that are inside our window of interest, copy corresponding matches,reads,qualities
while(mIt >= mItBegin && _min((*mIt).beginPos,(*mIt).endPos) + (TContigPos)options.maxHitLength + (TContigPos)options.windowBuff >= currentWindowEnd )
{
if( _max((*mIt).beginPos,(*mIt).endPos) + (TContigPos)options.windowBuff > currentWindowEnd )
{
TId id = length(tmpMatches);
appendValue(tmpMatches,*mIt);
tmpMatches[id].id = id;
tmpMatches[id].readId = id;
appendValue(tmpReads,fragmentStore.readSeqStore[(*mIt).readId]);
appendValue(tmpRs,fragmentStore.readStore[(*mIt).readId]);
if(!empty(readCounts))appendValue(tmpReadCounts,readCounts[(*mIt).readId]);
appendValue(tmpQualities,fragmentStore.alignQualityStore[(*mIt).readId]);
appendValue(tmpReadCigars,readCigars[(*mIt).readId]);
// appendValue(tmpReadClips,TPair(0,0));
appendValue(tmpReadClips,readClips[(*mIt).readId]);
maxCoord = std::max(maxCoord, (int)std::max(mIt->beginPos, mIt->endPos));
minCoord = std::min(minCoord, (int)std::min(mIt->beginPos, mIt->endPos));
}
--mIt;
}
// Write minimal and maximal coordinate from reads in this window to options.minCoord/options.maxCoord.
if (minCoord != std::numeric_limits<int>::max())
options.minCoord = minCoord;
if (maxCoord != std::numeric_limits<int>::min())
options.maxCoord = maxCoord;
if(options._debugLevel > 1)
std::cout << length(tmpMatches)<<" matches left over from previous window.\n";
}
// little helper
template<typename TMatch>
char
orientation(TMatch & match)
{
if(match.endPos > match.beginPos)
return 'F';
else
return 'R';
}
// discard reads stacking up, give preference to high quality reads
template<typename TFragmentStore, typename TSize, typename TOptions>
void
applyPileupCorrection(TFragmentStore &fragmentStore,
TSize arrayBeginPos,
TSize arrayEndPos,
TOptions &options)
{
typedef StringSet<String<Dna5Q>, Owner<ConcatDirect<> > > TFalseType;
typedef typename TFragmentStore::TAlignedReadStore TMatches;
//typedef typename Value<TMatches>::Type TMatch;
typedef typename TFragmentStore::TAlignQualityStore TMatchQualities;
//typedef typename Value<TMatchQualities>::Type TMatchQuality;
typedef typename TFragmentStore::TReadSeqStore TReads;
//typedef typename Value<TReads>::Type TRead;
//typedef typename TFragmentStore::TContigStore TGenomeSet;
//typedef typename Value<TGenomeSet>::Type TGenome;
typedef typename TFragmentStore::TContigPos TContigPos;
typedef typename Iterator<TMatches,Standard>::Type TMatchIterator;
if(IsSameType<TReads, TFalseType >::VALUE)
std::cout << "Hier stimmt was nciht. strinsetspec concat direct\n";
if(options._debugLevel > 0) std::cout << arrayEndPos - arrayBeginPos << " matches subject to pile up correction." << std::endl;
//CharString str = "pileBef";
//_dumpMatches(fragmentStore, str);
if(options.orientationAware)
::std::sort(iter(fragmentStore.alignedReadStore, arrayBeginPos, Standard()),
iter(fragmentStore.alignedReadStore, arrayEndPos, Standard()),
LessGStackOaMQ<TMatches,TMatchQualities>(fragmentStore.alignQualityStore));
else
::std::sort(iter(fragmentStore.alignedReadStore, arrayBeginPos, Standard()),
iter(fragmentStore.alignedReadStore, arrayEndPos, Standard()),
LessGStackMQ<TMatches,TMatchQualities>(fragmentStore.alignQualityStore));
TMatchIterator matchIt = iter(fragmentStore.alignedReadStore, arrayBeginPos, Standard());
TMatchIterator matchRangeEnd = iter(fragmentStore.alignedReadStore, arrayEndPos, Standard());
TMatchIterator matchItKeep = matchIt;
while(matchIt != matchRangeEnd)
{
TContigPos currentBegin = _min((*matchIt).beginPos,(*matchIt).endPos);
TContigPos currentEnd = _max((*matchIt).beginPos,(*matchIt).endPos);
unsigned currentSeqno = (*matchIt).contigId;
char currentOrientation = orientation(*matchIt);
unsigned currPile = 0;
while(matchIt != matchRangeEnd
&& (*matchIt).contigId == currentSeqno
&& _min((*matchIt).beginPos,(*matchIt).endPos) == currentBegin
&& _max((*matchIt).beginPos,(*matchIt).endPos) == currentEnd
&& (!options.orientationAware || orientation(*matchIt) == currentOrientation)
&& currPile < options.maxPile)
{
*matchItKeep = *matchIt;
++currPile;
++matchIt;
++matchItKeep;
}
//if(matchRangeEnd > matchItEnd) ::std::cerr <<"neeeeeeee\n";
while(matchIt != matchRangeEnd
&& (*matchIt).contigId == currentSeqno
&& _min((*matchIt).beginPos,(*matchIt).endPos) == currentBegin
&& _max((*matchIt).beginPos,(*matchIt).endPos) == currentEnd
&& (!options.orientationAware || orientation(*matchIt) == currentOrientation))
++matchIt;
}
if(options._debugLevel > 0) std::cout << matchIt - matchItKeep << " matches discarded." << std::endl;
resize(fragmentStore.alignedReadStore,matchItKeep - begin(fragmentStore.alignedReadStore,Standard()));
// ::std::cout << "numMatches = " << length(fragmentStore.alignedReadStore) << ::std::endl;
SEQAN_ASSERT_LEQ(length(fragmentStore.alignedReadStore), length(fragmentStore.alignQualityStore));
SEQAN_ASSERT_EQ(length(fragmentStore.readSeqStore), length(fragmentStore.alignQualityStore));
// str="pileAft";
// _dumpMatches(fragmentStore,str);
}
// average quality of read is kept as extra info for each match
// used for prioritization in pile up correction
template<typename TFragmentStore, typename TSize, typename TOptions>
void
addReadQualityToMatches(TFragmentStore &fragmentStore,
TSize arrayBeginPos,
TSize arrayEndPos,
TOptions &)
{
typedef typename TFragmentStore::TAlignedReadStore TMatches;
//typedef typename Value<TMatches>::Type TMatch;
typedef typename TFragmentStore::TReadSeqStore TReads;
typedef typename Value<TReads>::Type TRead;
typedef typename Iterator<TMatches, Standard>::Type TIterator;
TIterator it = iter(fragmentStore.alignedReadStore, arrayBeginPos, Standard());
TIterator itEnd = iter(fragmentStore.alignedReadStore, arrayEndPos, Standard());
int avgRQ;
for (; it != itEnd; ++it)
{
TRead const &read = fragmentStore.readSeqStore[(*it).readId];
avgRQ = 0;
for(unsigned i = 0; i < length(read); ++i)
avgRQ += (int) getQualityValue(read[i]) ;
// watch out, this is new: use mapping quality if given
if((fragmentStore.alignQualityStore[(*it).id]).score == 0 || (char)(avgRQ/length(read))<(fragmentStore.alignQualityStore[(*it).id]).score)
(fragmentStore.alignQualityStore[(*it).id]).score = (char)(avgRQ/length(read));
}
}
// checks whether an alignment has indels
template<typename TValue, typename TSpec>
bool alignmentHasIndel(Align<TValue,TSpec> &align)
{
typedef Align<TValue,TSpec> TAlign;
typedef typename Row<TAlign>::Type TAlignRow;
typedef typename Iterator<TAlignRow>::Type TAlignIterator;
bool hasIndel = false;
for(unsigned i = 0; !hasIndel && i < length(rows(align)); ++i)
{
TAlignIterator it = begin(row(align,i));
TAlignIterator itEnd = end(row(align,i));
while (it != itEnd)
{
if(isGap(it))
{
hasIndel = true;
break;
}
++it;
}
}
return hasIndel;
}
// perform read clipping
template<typename TFragmentStore, typename TReadClips, typename TSize, typename TOptions>
void
clipReads(TFragmentStore &fragmentStore,
TReadClips &readClips,
TSize arrayBeginPos,
TSize arrayEndPos,
TOptions &options)
{
typedef typename TFragmentStore::TAlignedReadStore TMatches;
typedef typename Value<TMatches>::Type TMatch;
typedef typename TFragmentStore::TAlignQualityStore TAlignQualityStore; // TMatchQualities
typedef typename Value<TAlignQualityStore>::Type TAlignQuality;
typedef typename TFragmentStore::TReadSeqStore TReads;
typedef typename Value<TReads>::Type TRead;
typedef typename Iterator<TMatches, Standard>::Type TIterator;
typedef typename TFragmentStore::TContigSeq TContigSeq; // TGenome
typedef typename Position<TContigSeq>::Type TContigPos;
TIterator it = iter(fragmentStore.alignedReadStore, arrayBeginPos, Standard());
TIterator itEnd = iter(fragmentStore.alignedReadStore, arrayEndPos, Standard());
Align<TRead, ArrayGaps> align;
resize(rows(align), 2);
#ifdef SNPSTORE_DEBUG
bool extraV = true;
#endif
Score<int> scoreType = Score<int>(0, -999, -1001, -1000);
if(length(readClips) < (arrayEndPos-arrayBeginPos)) ::std::cout << length(readClips) << " readclips but " << (arrayEndPos-arrayBeginPos) << " many reads.\n";
TContigSeq &currGenome = fragmentStore.contigStore[0].seq;
int kickout = 0;
for (; it != itEnd; ++it)
{
TMatch &m = *it;
TRead &read = fragmentStore.readSeqStore[m.readId];
int clipLeft = readClips[m.readId].i1;
int clipRight = readClips[m.readId].i2;
TContigPos beginPos = (m.beginPos < m.endPos ) ? m.beginPos : m.endPos;
TContigPos endPos = (m.beginPos > m.endPos ) ? m.beginPos : m.endPos;
TAlignQuality &aliQ = fragmentStore.alignQualityStore[m.id];
#ifdef SNPSTORE_DEBUG
TContigPos beginBefore = beginPos;
#endif
if(m.id != m.readId) ::std::cout << "match id != readId \n";
if(clipLeft+clipRight > (int)length(read) || clipLeft > (int)length(read) || clipRight > (int)length(read))
{
if(options._debugLevel > 0)::std::cout << "WARNING: clipLeft+clipRight > readLen!!\n";
#ifdef SNPSTORE_DEBUG
::std::cout << "readlength = "<<length(read)<< " \n";
::std::cout << "readId = "<<m.readId << "id=" << m.id << " \n";
::std::cout << "clipLeft = " << clipLeft << " clipRight = "<<clipRight << "\n";
::std::cout << "read=" << read << std::endl;
::std::cout << "beginPos=" << beginPos << std::endl;
::std::cout << "endPos=" << endPos << std::endl;
#endif
clipLeft = length(read);
clipRight = 0;
}
#ifdef SNPSTORE_DEBUG
::std::cout << "readlength = "<<length(read)<< " \n";
::std::cout << "readId = "<<m.readId << "id=" << m.id << " \n";
::std::cout << "clipLeft = " << clipLeft << " clipRight = "<<clipRight << "\n";
::std::cout << "read=" << read << std::endl;
::std::cout << "beginPos=" << beginPos << std::endl;
::std::cout << "endPos=" << endPos << std::endl;
#endif
if(clipLeft > 0 || clipRight > 0)
{
// if(extraV) ::std::cout << "clipLeft = " << clipLeft << " clipRight = "<<clipRight << std::endl;
if((int)length(read)-clipLeft-clipRight < options.minClippedLength)
{
if(options._debugLevel > 1 )
::std::cout << "Discarded: "<<read<<" at position "<< beginPos <<"\n";
m.endPos = m.beginPos = 0; // "mask" read
read = "";
++kickout;
continue;
}
// adjust read sequence
read = infix(read,clipLeft,length(read)-clipRight);
// upate avg read quality
int avgRQ = 0;
for(unsigned i = 0; i < length(read); ++i)
avgRQ += (int) getQualityValue(read[i]) ;
aliQ.score = (char)(avgRQ/length(read));
// if(extraV) ::std::cout << "aliQ.score = " << (int)aliQ.score << ::std::endl;
//do semi-global alignment
assignSource(row(align, 0), read);
assignSource(row(align, 1), infix(currGenome, beginPos, endPos));
if ((*it).endPos < (*it).beginPos)
reverseComplement(source(row(align, 1)));
int score = globalAlignment(align, scoreType, AlignConfig<false,true,true,false>(), Gotoh());
aliQ.errors = (unsigned) round((float)-score/1000);
#ifdef SNPSTORE_DEBUG
if(extraV) ::std::cout << align << std::endl;
if(extraV) ::std::cout << "aliQ.errors = " << (int) aliQ.errors << std::endl;
#endif
// transform first and last read character to genomic positions
if(aliQ.pairScore == 1)
{
unsigned viewPosReadFirst = toViewPosition(row(align, 0), 0);
unsigned viewPosReadLast = toViewPosition(row(align, 0), length(read) - 1);
unsigned genomePosReadFirst = toSourcePosition(row(align, 1), viewPosReadFirst);
unsigned genomePosReadLast = toSourcePosition(row(align, 1), viewPosReadLast);
// if(isGap(row(align,1),viewPosReadFirst))
// {
// std::cout << "bgein gap --> do nothing " << std::endl;
//
// }
if(isGap(row(align,1),viewPosReadLast))
{
genomePosReadLast--;
}
#ifdef SNPSTORE_DEBUG
if(extraV)
{ ::std::cout << "viewPosReadFirst = " << viewPosReadFirst << ::std::endl;
::std::cout << "viewPosReadLast = " << viewPosReadLast << ::std::endl;
::std::cout << "genomePosReadFirst = " << genomePosReadFirst << ::std::endl;
}
#endif
if(m.beginPos < m.endPos) // forward
{
endPos = beginPos + (genomePosReadLast + 1);
beginPos += genomePosReadFirst;
}
else
{
beginPos = endPos - genomePosReadLast - 1;
endPos = endPos - genomePosReadFirst;
}
if(!alignmentHasIndel(align)) aliQ.pairScore = 0;
}
else
{
if(m.beginPos < m.endPos) // forward
{
endPos -= clipRight;
beginPos += clipLeft;
}
else
{
endPos -= clipLeft;
beginPos += clipRight;
}
}
// set genomic positions
if(m.beginPos < m.endPos) // forward
{
m.endPos = endPos;
m.beginPos = beginPos;
}
else
{
m.endPos = beginPos;
m.beginPos = endPos;
}
if(beginPos > 300000000 || endPos > 300000000)
{
::std::cout << "bgein groesser 300mio neu beginPos = "<< beginPos << " endpos=" << endPos << ::std::endl;
#ifdef SNPSTORE_DEBUG
::std::cout << "WARNING: clipLeft+clipRight > readLen!!??\n";
::std::cout << "readId = "<<m.readId << "id=" << m.id << " \n";
::std::cout << "clipLeft = " << clipLeft << " clipRight = "<<clipRight << "\n";
::std::cout << "read=" << read << std::endl;
::std::cout << "beginPos before=" << beginBefore << std::endl;
::std::cout << "beginPos=" << beginPos << std::endl;
::std::cout << "endPos=" << endPos << std::endl;
#endif
}
}
}
if(options._debugLevel > 0)
::std::cout << kickout <<" reads too short after clipping, discarding!\n";
}
template<typename TTable>
void
printHetTable(TTable & hetTable)
{
cout << "Printing het table:" << std::endl;
for (int n1=0; n1<256; ++n1)
{
for (int n2=0; n2<256; ++n2)
{
cout << hetTable[n1<<8|n2] << "\t";
}
cout << std::endl;
}
cout << std::endl;
}
template<typename TTmpReads, typename TTmpMatches, typename TTmpQualities, typename TStr>
void
_dumpMatches(TTmpReads & reads, TTmpMatches & matches, TTmpQualities & qualities, TStr str)
{
std::cout << "Length of matches = " << length(matches) << "\n";
std::cout << "Length of reads = " << length(reads) << "\n";
std::cout << "Length of matchqs = " << length(qualities) << "\n";
for(unsigned i = 0 ; i < length(matches); ++i)
{
char ori = (matches[i].beginPos < matches[i].endPos) ? 'F' : 'R';
std::cout << "--"<<str<<"Match number " << i << ":\n";
std::cout << "--"<<str<<"MatchId = " << matches[i].id << "\n";
std::cout << "--"<<str<<"ReadId = " << matches[i].readId << "\n";
std::cout << "--"<<str<<"ContigId = " << matches[i].contigId << std::flush << "\n";
std::cout << "--"<<str<<"gBegin = " << _min(matches[i].beginPos, matches[i].endPos) << "\n";
std::cout << "--"<<str<<"gEnd = " << _max(matches[i].beginPos, matches[i].endPos) << "\n";
std::cout << "--"<<str<<"orient = " << ori << std::flush << std::endl;
if(length(qualities) > matches[i].id)
{
std::cout << "--"<<str<<"EditDist = " << (int) qualities[matches[i].id].errors << "\n";
std::cout << "--"<<str<<"AvgQ = " << (int) qualities[matches[i].id].score << "\n";
}
std::cout << "--"<<str<<"Readseq = " << reads[matches[i].readId] << std::flush << "\n";
}
}
//////////////////////////////////////////////////////////////////////////////
// Main read mapper function
template <typename TSpec>
int detectSNPs(SNPCallingOptions<TSpec> &options)
{
typedef FragmentStore<SnpStoreSpec_> TFragmentStore;
//typedef typename TFragmentStore::TReadSeq TReadSeq; // TRead
typedef typename TFragmentStore::TContigSeq TContigSeq; // TGenome
//typedef typename Position<TReadSeq>::Type TReadPos; // TPos
//typedef typename TFragmentStore::TReadPos TReadPos; // TPos
//typedef typename Position<TContigSeq>::Type TContigPos; // TContigPos
typedef typename TFragmentStore::TContigPos TContigPos;
typedef typename TFragmentStore::TAlignedReadStore TAlignedReadStore; // TMatches
typedef typename TFragmentStore::TAlignQualityStore TAlignQualityStore; // TMatchQualities
typedef typename TFragmentStore::TReadStore TReadStore; // TReadSet
typedef typename TFragmentStore::TReadSeqStore TReadSeqStore; // TReadSet
typedef typename TFragmentStore::TContigStore TContigStore; // TGenomeSet
typedef typename Value<TContigStore>::Type TContig;
typedef TContigSeq TGenome;
typedef StringSet<TGenome> TGenomeSet;
typedef String<String<TContigPos > > TPositions;
typedef typename Iterator<String<TContigPos > >::Type TPosIterator;
typedef typename Size<TGenomeSet>::Type TGenomeSetSize;
typedef ::std::map<CharString, TGenomeSetSize> TGenomeMap;
//typedef typename TGenomeMap::iterator TMapIter;
typedef String<unsigned> TReadCounts;
typedef String<Pair<int,int> > TReadClips;
typedef StringSet<String<Pair<char,int> > > TReadCigars;
TGenomeSet genomes;
StringSet<CharString> genomeFileNameList; // filenamen
StringSet<CharString> genomeNames; // todo: raus
TGenomeMap gIdStringToIdNumMap; // name to id
TPositions positions; // list of positions to inspect
// dump configuration in verbose mode
if (options._debugLevel >= 1)
{
::std::cerr << "___SETTINGS____________" << ::std::endl;
::std::cerr << "Genome file: \t" << options.genomeFName << ::std::endl;
::std::cerr << "Read files: \t" << options.readFNames[0] << ::std::endl;
for(unsigned i = 1; i < length(options.readFNames); ++i)
::std::cerr << " \t" << options.readFNames[i] << ::std::endl;
/* if(options.inputFormat == 1)
{
::std::cerr << "Quality files: \t" << qualityFNames[0] << ::std::endl;
for(unsigned i = 1; i < length(qualityFNames); ++i)
::std::cerr << " \t" << qualityFNames[i] << ::std::endl;
}*/::std::cerr << "MaxPile: \t" << options.maxPile << ::std::endl;
if(options.laneSpecificMaxPile)::std::cerr << "Lane specific: \tYES" << ::std::endl;
else ::std::cerr << "Lane specific: \tNO" << ::std::endl;
::std::cerr << "MinCoverage: \t" << options.minCoverage << ::std::endl;
if(options.method == 0)
{
::std::cerr << "MinMutThreshold: \t" << options.minMutT << ::std::endl;
::std::cerr << "MinPercentageThreshold: \t" << options.percentageT << ::std::endl;
::std::cerr << "MinQualityThreshold: \t" << options.avgQualT << ::std::endl;
}
else
{
::std::cerr << "MinMappingQuality: \t" << options.minMapQual << ::std::endl;
}
if(options.outputIndel != "")
{
::std::cerr << "IndelCountThreshold: \t" << options.indelCountThreshold << ::std::endl;
::std::cerr << "IndelPercentageThreshold: \t" << options.indelPercentageT << ::std::endl;
::std::cerr << "IndelWindow: \t" << options.indelWindow << ::std::endl;
}
::std::cerr << ::std::endl;
}
//////////////////////////////////////////////////////////////////////////////
// Step 1: Determine genome file type and load genomes
SEQAN_PROTIMESTART(load_time);
int result = getGenomeFileNameList(genomeFileNameList, options);
if(result == CALLSNPS_GENOME_FAILED || !loadGenomes(genomes, genomeFileNameList, gIdStringToIdNumMap, genomeNames))
{
::std::cerr << "Failed to open genome file " << options.genomeFName << ::std::endl;
return result;
}
//////////////////////////////////////////////////////////////////////////////
// Step 2: Load fragmentStore.readSeqStore and fragmentStore.alignedReadStore
// open read files and store open file pointers
String<int> highestChrId;
resize(highestChrId,length(options.readFNames),0);
vector< ::std::fstream* > readFileStreams;
readFileStreams.resize(length(options.readFNames));
for (unsigned i = 0; options.inputFormat == 0 && i < length(options.readFNames); ++i)
{
readFileStreams[i] = new fstream(toCString(options.readFNames[i]), ios_base::in | ios::binary);
if(!(*(readFileStreams[i])).is_open())
{
::std::cerr << "Failed to open read file " << options.readFNames[i] << ::std::endl;
return CALLSNPS_GFF_FAILED;
}
}
// Open all input SAM and BAM files.
std::vector<BamFileIn *> bamFileIns;
if (options.inputFormat >= 1) // 1 -- SAM, 2 -- BAM
for (unsigned i = 0; i < length(options.readFNames); ++i)
{
bamFileIns.push_back(new BamFileIn());
//std::cout << "Opening SAM/BAM file\n";
if (!open(*bamFileIns.back(), toCString(options.readFNames[i])))
{
std::cerr << "[ERROR] Could not open BAM file" << options.readFNames[i] << "\n";
return 1;
}
}
// Allocate one BamAlignmentRecord for each input BAM file.
std::vector<BamAlignmentRecord> records(length(options.readFNames));
/////////////////////////////////////////////////////////////////////
// open out file streams and store open file pointers
::std::ofstream snpFileStream;
if (options.outputSNP != "")
{
// prepare lookup tables for maq statistics
if (options.method == 1 )
{
computeCnks(options.cnks,options.fks,options);
#ifdef CORRECTED_HET
if(options.correctedHetTable) options.priorHetQ = computeHetTable(options.hetTable,options); // corrected het table computed with normal distribution
else
#endif
options.priorHetQ = computeHetTable(options.hetTable,options,MaqMethod()); // original Maq method
//printHetTable(options.hetTable);
//printHetTable(options.hetTable2);
}
snpFileStream.open(toCString(options.outputSNP),::std::ios_base::out);
if(!snpFileStream.is_open())
return CALLSNPS_OUT_FAILED;
snpFileStream << "#" << (options.programCall).str() << "\n";
if(options.outputFormat < 2)
{
if(options.orientationAware)
snpFileStream << "#chr\tpos\tref\t[A+]\t[C+]\t[G+]\t[T+]\t[A-]\t[C-]\t[G-]\t[T-]\tcov\tcall";
else
snpFileStream << "#chr\tpos\tref\tA\tC\tG\tT\tcov\tcall";
//if(options.method==1)
snpFileStream << "\tquality\tsnpQ\n";
//else
// file <<"\n";
}
}
::std::ofstream indelFileStream;
if (options.outputIndel != "")
{
indelFileStream.open(toCString(options.outputIndel),::std::ios_base::out);
if(!indelFileStream.is_open())
return CALLSNPS_OUT_FAILED;
}
// ::std::ofstream cnvFileStream;
// if (*options.outputCNV != 0)
// {
// cnvFileStream.open(options.outputCNV,::std::ios_base::out);
// if(!cnvFileStream.is_open())
// return CALLSNPS_OUT_FAILED;
// }
::std::ofstream posFileStream;
if(options.inputPositionFile != "")
{
resize(positions,length(genomeNames));
result = loadPositions(positions,gIdStringToIdNumMap, toCString(options.inputPositionFile),options);
if(result != 0)
{
::std::cerr << "Failed to read position file " << options.inputPositionFile << ::std::endl;
return result;
}
posFileStream.open(toCString(options.outputPosition),::std::ios_base::out);
if(!posFileStream.is_open())
return CALLSNPS_OUT_FAILED;
if(options.orientationAware)
posFileStream << "#chr\tpos\t[A+]\t[C+]\t[G+]\t[T+]\t[gap+]\t[A-]\t[C-]\t[G-]\t[T-]\t[gap-]\n";
else
posFileStream << "#chr\tpos\tA\tC\tG\tT\tgap\n";
}
/////////////////////////////////////////////////////////////////////////////
// helper variables
Pair<int,int> zeroPair(0,0);
bool positionStatsOnly = (options.outputSNP == "" && options.outputPosition != "");
TPosIterator inspectPosIt, inspectPosItEnd;
// The iteration of the positions entries is convoluted with the rest of the code. We create a
// fake positions entry of the correct size (length(genomes)) but for positionStatsOnly to false
// if no positions are given.
if (empty(positions))
{
resize(positions, length(genomes));
positionStatsOnly = false; // ignor einspectPosIt below
}
bool firstCall = true;
/////////////////////////////////////////////////////////////////////////////
// Start scanning for SNPs/indels
// for each chromosome
for(unsigned i=0; i < length(genomes); ++i)
{
//std::cout << genomeNames[i] << "\n";
inspectPosIt = begin(positions[i], Standard());
inspectPosItEnd = end(positions[i], Standard());
if (positionStatsOnly && inspectPosIt == inspectPosItEnd)
continue;
// parse matches batch by batch
TContigPos currentWindowBegin = 0;
if(options._debugLevel > 0) ::std::cout << "Scanning genome #" << i << " ..." << ::std::endl;
// containers for those matches that overlap two windows
TAlignedReadStore tmpMatches;
TAlignQualityStore tmpQualities;
TReadStore tmpRs;
TReadSeqStore tmpReads;
TReadCounts tmpReadCounts;
TReadClips tmpReadClips;
TReadCigars tmpReadCigars;
options.minCoord = std::numeric_limits<unsigned>::max();
options.maxCoord = 0;
// snp calling is done for all positions between windowBegin and windowEnd
while(currentWindowBegin < (TContigPos)length(genomes[i]))
{
TContigPos currentWindowEnd = currentWindowBegin + options.windowSize;
if(currentWindowEnd > (TContigPos)length(genomes[i])) currentWindowEnd = (TContigPos)length(genomes[i]);
if(positionStatsOnly && !(currentWindowBegin <= *inspectPosIt && *inspectPosIt< currentWindowEnd))
{
SEQAN_ASSERT_LEQ(currentWindowBegin,*inspectPosIt);
clear(tmpReads); clear(tmpRs); clear(tmpMatches); clear(tmpQualities);
clear(tmpReadClips); clear(tmpReadCounts); clear(tmpReadCigars);
currentWindowBegin = currentWindowEnd;
continue;
}
if(options._debugLevel > 0)
::std::cout << "Sequence number " << i << " window " << currentWindowBegin << ".." << currentWindowEnd << "\n";
TFragmentStore fragmentStore;
TReadCounts readCounts; // Count number of reads that are identical to the given one. Useful for micro RNA data where there were millions of identical reads. Must be in GFF input, not supported for SAM input.
TReadClips readClips; // Soft clipping information and/or clipping information from GFF/SAM tag. Clipping is postponed after pileup correction.
TReadCigars readCigars; // Currently only stored for split-mapped reads. Split-mapped reads need special handling, especially for realignment.
// add the matches that were overlapping with this and the last window (copied in order to avoid 2 x makeGlobal)
if(!empty(tmpMatches))
{
resize(fragmentStore.alignQualityStore,length(tmpMatches));
resize(fragmentStore.alignedReadStore,length(tmpMatches));
resize(fragmentStore.readSeqStore,length(tmpMatches));
resize(fragmentStore.readStore,length(tmpMatches));
if(!empty(tmpReadClips))resize(readClips,length(tmpMatches));
if(!empty(tmpReadCounts)) resize(readCounts,length(tmpMatches));
if(!empty(tmpReadCigars))resize(readCigars,length(tmpMatches));
arrayMoveForward(begin(tmpQualities,Standard()), end(tmpQualities,Standard()), begin(fragmentStore.alignQualityStore,Standard()));
arrayMoveForward(begin(tmpMatches,Standard()), end(tmpMatches,Standard()), begin(fragmentStore.alignedReadStore,Standard()));
arrayMoveForward(begin(tmpReads,Standard()), end(tmpReads,Standard()), begin(fragmentStore.readSeqStore,Standard()));
arrayMoveForward(begin(tmpRs,Standard()), end(tmpRs,Standard()), begin(fragmentStore.readStore,Standard()));
if(!empty(tmpReadCounts)) arrayMoveForward(begin(tmpReadCounts,Standard()), end(tmpReadCounts,Standard()), begin(readCounts,Standard()));
if(!empty(tmpReadCigars)) arrayMoveForward(begin(tmpReadCigars,Standard()), end(tmpReadCigars,Standard()), begin(readCigars,Standard()));
if(!empty(tmpReadClips)) arrayMoveForward(begin(tmpReadClips,Standard()), end(tmpReadClips,Standard()), begin(readClips,Standard()));
#ifdef SNPSTORE_DEBUG
CharString strstr = "afterCopyInFrag";
_dumpMatches(fragmentStore,strstr);
#endif
}
// parse matches for current window
if(options._debugLevel > 0)
::std::cout << "Parsing reads up to position " << currentWindowEnd << "...\n";
for(unsigned j = 0; j < length(options.readFNames); ++j)
{
unsigned sizeBefore = length(fragmentStore.alignedReadStore);
// currently only gff supported
if (options.inputFormat == 0) // GFF
result = readMatchesFromGFF_Batch(readFileStreams[j], fragmentStore, readCounts, readClips,
readCigars, genomes[i], gIdStringToIdNumMap,
i, currentWindowBegin, currentWindowEnd, highestChrId[j], options);
if (options.inputFormat >= 1) // SAM
result = readMatchesFromSamBam_Batch(*bamFileIns[j], records[j], fragmentStore, readCounts, readClips,
readCigars, genomes[i], gIdStringToIdNumMap,
i, currentWindowBegin, currentWindowEnd, highestChrId[j], options, firstCall);
firstCall = false;
if (result == CALLSNPS_GFF_FAILED)
{
std::cerr << "Failed to open read file " << options.readFNames[j] << ::std::endl;
std::cerr << "or reads are not sorted correctly. " << ::std::endl;
return result;
}
if (result > 0)
return result;
if (options._debugLevel > 0)
std::cout << "parsed reads of file " << j << "\n";
// store average quality of each read
addReadQualityToMatches(fragmentStore,sizeBefore,(unsigned)length(fragmentStore.alignedReadStore),options);
// do pile up correction if lane-based. lane-specific pileup correction not really used
if(options.maxPile != 0 && options.laneSpecificMaxPile)
applyPileupCorrection(fragmentStore,sizeBefore,(unsigned)length(fragmentStore.alignedReadStore),options);
}
if (options._debugLevel > 1) // number of reads currently in memory
::std::cerr << lengthSum(fragmentStore.readSeqStore) << " bps of " << length(fragmentStore.readSeqStore) << " reads in memory." << ::std::endl;
// do merged pile up correction
if(options.maxPile != 0 && !options.laneSpecificMaxPile)
applyPileupCorrection(fragmentStore,(unsigned)0,(unsigned)length(fragmentStore.alignedReadStore),options);
// these were set while parsing matches, first and last position of parsed matches
// TContigPos startCoord = options.minCoord;// can be < currentWindowBegin
// TContigPos endCoord = options.maxCoord; // can be > currentWindoEnd
// check
TContigPos startCoord = _max((int)options.minCoord-options.realignAddBorder,0);// can be < currentWindowBegin
TContigPos endCoord = _min(options.maxCoord+options.realignAddBorder,length(genomes[i])); // can be > currentWindoEnd
if(!empty(fragmentStore.alignedReadStore))
{
//initial values of min and max coords for next round are set here
if(currentWindowEnd != (TContigPos)length(genomes[i]))
{
clear(tmpMatches);
clear(tmpQualities);
clear(tmpRs);
clear(tmpReads);
clear(tmpReadCounts);
clear(tmpReadClips);
clear(tmpReadCigars);
copyNextWindowMatchesAndReads(fragmentStore,readCounts,readCigars,readClips,tmpReadCounts,tmpReads,tmpRs,tmpMatches,tmpQualities,tmpReadClips,tmpReadCigars,i,currentWindowEnd,options);
#ifdef SNPSTORE_DEBUG
CharString strstr = "afterCopyInTmp";
_dumpMatches(tmpReads,tmpMatches,tmpQualities,strstr);
#endif
}
// ::std::cout << "Min = " << options.minCoord << " Max = " << options.maxCoord << std::endl;
// ::std::cout << "Min = " << startCoord << " Max = " << endCoord << std::endl;
// coordinates are relative to current chromosomal window (segment)
transformCoordinates(fragmentStore,startCoord,options);
// set the current chromosomal segment as contig sequence
TContig conti;
conti.seq = infix(genomes[i],startCoord,endCoord);
appendValue(fragmentStore.contigStore, conti, Generous() );
appendValue(fragmentStore.contigNameStore, genomeNames[i], Generous() );// internal id is always 0
// clip Reads if clipping is switched on and there were clip tags in the gff file
if((!options.dontClip && options.clipTagsInFile) || options.softClipTagsInFile)
{
options.useBaseQuality = false; // activate "average read quality"-mode for snp calling, low quality bases should be clipped anyway
clipReads(fragmentStore,readClips,(unsigned)0,(unsigned)length(fragmentStore.alignedReadStore),options);
}
// check for indels
if (options.outputIndel != "")
{
if(options._debugLevel > 1) ::std::cout << "Check for indels..." << std::endl;
if(!options.realign) dumpShortIndelPolymorphismsBatch(fragmentStore, readCigars, fragmentStore.contigStore[0].seq, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, indelFileStream, options);
}
// // check for CNVs
// if (*options.outputCNV != 0)
// dumpCopyNumberPolymorphismsBatch(fragmentStore, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, cnvFileStream, options);
#ifdef SNPSTORE_DEBUG
CharString strstr = "test";
// _dumpMatches(fragmentStore, strstr );
#endif
if (options.outputSNP != "")
{
if(options._debugLevel > 1) ::std::cout << "Check for SNPs..." << std::endl;
if(options.realign)
dumpVariantsRealignBatchWrap(fragmentStore, readCigars, readCounts, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, snpFileStream,indelFileStream,options);
else
dumpSNPsBatch(fragmentStore, readCigars, readCounts, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, snpFileStream,options);
}
if(positionStatsOnly)
{
if(options._debugLevel > 1) ::std::cout << "Dumping info for query positions..." << std::endl;
if(options.realign)
dumpPositionsRealignBatchWrap(fragmentStore, inspectPosIt, inspectPosItEnd, readCigars, readCounts, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, posFileStream, options);
else dumpPosBatch(fragmentStore, inspectPosIt, inspectPosItEnd, readCigars, readCounts, genomeNames[i], startCoord, currentWindowBegin, currentWindowEnd, posFileStream, options);
}
}
// else
// {
if(positionStatsOnly && !empty(positions))
{
while(inspectPosIt != inspectPosItEnd && *inspectPosIt < currentWindowEnd)
{
if(options.orientationAware)
posFileStream << genomeNames[i] << '\t' << *inspectPosIt + options.positionFormat << "\t0\t0\t0\t0\t0\t0\t0\t0\t0\t0\t0" << std::endl;
else
posFileStream << genomeNames[i] << '\t' << *inspectPosIt + options.positionFormat << "\t0\t0\t0\t0\t0\t0" << std::endl;
++inspectPosIt;
}
}
// }
currentWindowBegin = currentWindowEnd;
}
}
if (options.outputSNP != "")
snpFileStream.close();
if (options.outputIndel != "")
indelFileStream.close();
if (options.outputPosition != "")
posFileStream.close();
// if (options.outputCNV != "")
// cnvFileStream.close();
return 0;
}
// log file to keep track of happenings
template <typename TSpec>
int writeLogFile(int argc, const char *argv[], SNPCallingOptions<TSpec> &options)
{
::std::ofstream logfile;
logfile.open(toCString(options.outputLog), ::std::ios_base::out | ::std::ios_base::trunc);
if (!logfile.is_open())
{
::std::cerr << "Failed to open log file" << ::std::endl;
return 1;
}
logfile << "#call" << std::endl;
for (int i = 0; i < argc; ++i)
logfile << argv[i] << " ";
logfile << std::endl;
logfile << "#files" << ::std::endl;
logfile << "Genome=\"" << options.genomeFName << "\""<<::std::endl;
logfile << "Reads=\"" << options.readFNames[0];
for(unsigned i = 1; i < length(options.readFNames); ++i)
logfile << " " << options.readFNames[i] << ::std::endl;
logfile << "\"" << std::endl;
if(options.outputSNP != "")
{
logfile << "OutputSnp=\"" << CharString(options.outputSNP) << "\"" << ::std::endl;
}
if(options.outputIndel != "")
{
logfile << "OutputIndel=\"" << CharString(options.outputIndel) << "\""<< ::std::endl;
}
logfile << "#settings" << ::std::endl;
logfile << "MaxPile=" << options.maxPile << ::std::endl;
logfile << "MinCov=" << options.minCoverage << ::std::endl;
logfile << "Method=" << options.method << ::std::endl;
if(options.method == 0)
{
logfile << "MinMutT=" << options.minMutT << ::std::endl;
logfile << "MinPercT=" << options.percentageT << ::std::endl;
logfile << "MinQualT=" << options.avgQualT << ::std::endl;
}
else
{
logfile << "MinMapQual=" << options.minMapQual << ::std::endl;
}
if(options.outputIndel != "")
{
logfile << "MinIndel=" << options.indelCountThreshold << ::std::endl;
logfile << "MinPercIndelT=" << options.indelPercentageT << ::std::endl;
}
logfile.close();
return 0;
}
template <typename TSpec>
ArgumentParser::ParseResult
parseCommandLine(SNPCallingOptions<TSpec> & options, int argc, char const ** argv)
{
// Setup ArgumentParser.
ArgumentParser parser("snp_store");
// Set short description, version, and date.
setShortDescription(parser, "SnpStore");
setCategory(parser, "Variant Detection");
setVersion(parser, SEQAN_APP_VERSION " [" SEQAN_REVISION "]");
setDate(parser, SEQAN_DATE);
// Define usage line and long description.
addUsageLine(parser, "[\\fIOPTIONS\\fP] <\\fIGENOME FILE\\fP> <\\fIALIGNMENT FILE\\fP> [<\\fIALIGNMENT FILE\\fP> ...]");
// We require two mandatory arguments: genome file and read file(s)
addArgument(parser, ArgParseArgument(seqan2::ArgParseArgument::INPUT_FILE, "GENOME"));
setValidValues(parser, 0, ".fa .fasta");
setHelpText(parser, 0, "A reference genome file.");
std::vector<std::string> alignmentFormats(BamFileIn::getFileExtensions());
alignmentFormats.push_back(".gff");
addArgument(parser, ArgParseArgument(seqan2::ArgParseArgument::INPUT_FILE, "ALIGNMENTS", true));
setValidValues(parser, 1, alignmentFormats);
setHelpText(parser, 1, "Read alignment file(s) sorted by genomic position.");
addDescription(parser, "SNP and Indel Calling in Mapped Read Data.");
addSection(parser, "Main Options");
addOption(parser, ArgParseOption("o", "output", "SNP output file (mandatory).", ArgParseArgument::OUTPUT_FILE));
setValidValues(parser, "output", ".vcf");
setRequired(parser, "output");
addOption(parser, ArgParseOption("osc", "only-successful-candidates", "Output only successfully called SNP candidates. Default: Output all candidates."));
addOption(parser, ArgParseOption("dc", "dont-clip", "Ignore clip tags in gff. Default: off."));
addOption(parser, ArgParseOption("mu", "multi", "Keep non-unique fragmentStore.alignedReadStore. Default: off."));
addOption(parser, ArgParseOption("hq", "hide-qualities", "Only show coverage (no qualities) in SNP output file. Default: off."));
addOption(parser, ArgParseOption("sqo", "solexa-qual-offset", "Base qualities are encoded as char value - 64 (instead of char - 33)."));
addOption(parser, ArgParseOption("id", "indel-file", "Output file for called indels in gff format. Default: off.", ArgParseArgument::OUTPUT_FILE));
setValidValues(parser, "indel-file", ".gff");
addOption(parser, ArgParseOption("m", "method", "Set method used for SNP calling either threshold based or Maq method.", ArgParseArgument::STRING));
setValidValues(parser, "method", "thresh maq");
setDefaultValue(parser, "method", "maq");
addOption(parser, ArgParseOption("mp", "max-pile", "Maximal number of matches allowed to pile up at the same genome position.", ArgParseArgument::INTEGER));
setMinValue(parser, "max-pile", "1");
setDefaultValue(parser, "max-pile", options.maxPile);
addOption(parser, ArgParseOption("mmp", "merged-max-pile", "Do pile up correction on merged lanes. Default: off."));
addOption(parser, ArgParseOption("mc", "min-coverage", "Minimal required number of reads covering a candidate position.", ArgParseArgument::INTEGER));
setMinValue(parser, "min-coverage", "1");
setDefaultValue(parser, "min-coverage", options.minCoverage);
addOption(parser, ArgParseOption("fc", "force-call", "Always call base if count is >= fc, ignore other parameters. Default: off.", ArgParseArgument::INTEGER));
setMinValue(parser, "force-call", "1");
setDefaultValue(parser, "force-call", options.forceCallCount);
addOption(parser, ArgParseOption("oa", "orientation-aware", "Distinguish between forward and reverse reads. Default: off."));
// addOption(parser, ArgParseOption("cnv", "output-cnv", "Name of CNV result file.", ArgParseArgument::OUTPUT_FILE));
// hideOption(parser, "cnv");
addOption(parser, ArgParseOption("op", "output-positions", "Name of positions output file.", ArgParseArgument::STRING));
hideOption(parser, "op");
addOption(parser, ArgParseOption("ip", "input-positions", "Name of positions input file.", ArgParseArgument::STRING));
hideOption(parser, "ip");
addOption(parser, ArgParseOption("mpr", "max-polymer-run", "Discard indels in homopolymer runs longer than mpr.", ArgParseArgument::INTEGER));
setMinValue(parser, "max-polymer-run", "0");
setDefaultValue(parser, "max-polymer-run", options.maxPolymerRun);
addOption(parser, ArgParseOption("dp", "diff-pos", "Minimal number of different read positions supporting the mutation.", ArgParseArgument::INTEGER));
setMinValue(parser, "diff-pos", "0");
setDefaultValue(parser, "diff-pos", options.minDifferentReadPos);
addOption(parser, ArgParseOption("eb", "exclude-border", "Exclude read positions within eb base pairs of read borders for SNV calling. Default: off.", ArgParseArgument::INTEGER));
setMinValue(parser, "exclude-border", "0");
setDefaultValue(parser, "exclude-border", options.excludeBorderPos);
addOption(parser, ArgParseOption("su", "suboptimal", "Keep suboptimal reads. Default: off"));
addOption(parser, ArgParseOption("re", "realign", "Realign reads around indel candidates. Default: off"));
addOption(parser, ArgParseOption("cq", "corrected-quality", "New quality calibration factor.", ArgParseArgument::DOUBLE));
setMinValue(parser, "corrected-quality", "0");
setDefaultValue(parser, "corrected-quality", options.newQualityCalibrationFactor);
hideOption(parser, "cq");
addOption(parser, ArgParseOption("pws", "parse-window-size", "Genomic window size for parsing reads (concerns memory consumption, choose smaller windows for higher coverage).", ArgParseArgument::INTEGER));
setMinValue(parser, "parse-window-size", "1");
setDefaultValue(parser, "parse-window-size", options.windowSize);
addOption(parser, ArgParseOption("reb", "realign-border", "Realign border.", ArgParseArgument::INTEGER));
setMinValue(parser, "realign-border", "0");
setMaxValue(parser, "realign-border", "10");
setDefaultValue(parser, "realign-border", options.realignAddBorder);
hideOption(parser, "reb");
addSection(parser, "SNP calling options");
addSection(parser, " Threshold method related");
addOption(parser, ArgParseOption("mm", "min-mutations", "Minimal number of observed mutations for mutation to be called.", ArgParseArgument::INTEGER));
setMinValue(parser, "min-mutations", "1");
setDefaultValue(parser, "min-mutations", options.minMutT);
addOption(parser, ArgParseOption("pt", "perc-threshold", "Minimal percentage of mutational base for mutation to be called.", ArgParseArgument::DOUBLE));
setMinValue(parser, "perc-threshold", "0");
setDefaultValue(parser, "perc-threshold", options.percentageT);
addOption(parser, ArgParseOption("mq", "min-quality", "Minimal average quality of mutational base for mutation to be called.", ArgParseArgument::DOUBLE));
setMinValue(parser, "min-quality", "0");
setDefaultValue(parser, "min-quality", options.avgQualT);
addSection(parser, " Maq method related");
addOption(parser, ArgParseOption("th", "theta", "Dependency coefficient.", ArgParseArgument::DOUBLE));
setMinValue(parser, "theta", "0");
setDefaultValue(parser, "theta", options.theta);
addOption(parser, ArgParseOption("hr", "hetero-rate", "Heterozygote rate.", ArgParseArgument::DOUBLE));
setMinValue(parser, "hetero-rate", "0");
setMaxValue(parser, "hetero-rate", "1");
setDefaultValue(parser, "hetero-rate", options.hetRate);
addOption(parser, ArgParseOption("mmq", "min-map-quality", "Minimum base call (mapping) quality for a match to be considered.", ArgParseArgument::INTEGER));
setMinValue(parser, "min-map-quality", "0");
setDefaultValue(parser, "min-map-quality", options.minMapQual);
addOption(parser, ArgParseOption("ch", "corrected-het", "Use amplification bias corrected distribution for heterozygotes. Default: off."));
addOption(parser, ArgParseOption("maf", "mean-alleleFreq", "Mean ref allele frequency in heterozygotes.", ArgParseArgument::DOUBLE));
setMinValue(parser, "mean-alleleFreq", "0");
setDefaultValue(parser, "mean-alleleFreq", options.meanAlleleFrequency);
addOption(parser, ArgParseOption("ac", "amp-cycles", "Number of amplification cycles.", ArgParseArgument::INTEGER));
setMinValue(parser, "amp-cycles", "0");
setDefaultValue(parser, "amp-cycles", options.amplificationCycles);
addOption(parser, ArgParseOption("ae", "amp-efficiency", "Polymerase efficiency, probability of amplification.", ArgParseArgument::DOUBLE));
setMinValue(parser, "amp-efficiency", "0");
setMaxValue(parser, "amp-efficiency", "1");
setDefaultValue(parser, "amp-efficiency", options.amplificationEfficiency);
addOption(parser, ArgParseOption("in", "initial-N", "Initial allele population size.", ArgParseArgument::INTEGER));
setMinValue(parser, "initial-N", "0");
setDefaultValue(parser, "initial-N", options.initialN);
addOption(parser, ArgParseOption("pht", "print-hetTable", "Print het table. Default: off."));
hideOption(parser, "pht");
addOption(parser, ArgParseOption("mec", "min-explained-column", "Minimum fraction of alignment column reads explained by genotype call.", ArgParseArgument::DOUBLE));
setMinValue(parser, "min-explained-column", "0");
setMaxValue(parser, "min-explained-column", "1");
setDefaultValue(parser, "min-explained-column", options.minExplainedColumn);
addSection(parser, "Indel calling options");
addOption(parser, ArgParseOption("it", "indel-threshold", "Minimal number of indel-supporting reads required for indel calling.", ArgParseArgument::INTEGER));
setMinValue(parser, "indel-threshold", "1");
setDefaultValue(parser, "indel-threshold", options.indelCountThreshold);
addOption(parser, ArgParseOption("ipt", "indel-perc-threshold", "Minimal ratio of indel-supporting/covering reads for indel to be called.", ArgParseArgument::DOUBLE));
setMinValue(parser, "indel-perc-threshold", "0");
setMaxValue(parser, "indel-perc-threshold", "1");
setDefaultValue(parser, "indel-perc-threshold", options.indelPercentageT);
addOption(parser, ArgParseOption("iqt", "indel-quality-thresh", "Minimal average quality of inserted base/deletion-neighboring bases for indel to be called.", ArgParseArgument::INTEGER));
setMinValue(parser, "indel-quality-thresh", "0");
setDefaultValue(parser, "indel-quality-thresh", options.indelQualityThreshold);
addOption(parser, ArgParseOption("bsi", "both-strands-indel", "Both strands need to be observed for indel to be called. Default: off."));
addOption(parser, ArgParseOption("iw", "indel-window", "Overlap window used for indel calling.", ArgParseArgument::INTEGER));
setMinValue(parser, "indel-window", "0");
setDefaultValue(parser, "indel-window", options.indelWindow);
hideOption(parser, "iw");
addOption(parser, ArgParseOption("ebi", "exclude-border-indel", "Same as option -eb but for indel candidates.", ArgParseArgument::INTEGER));
setMinValue(parser, "exclude-border-indel", "0");
setDefaultValue(parser, "exclude-border-indel", options.indelDepthMinOverlap);
addOption(parser, ArgParseOption("cws", "cnv-window-size", "CNV window size.", ArgParseArgument::INTEGER));
setMinValue(parser, "cnv-window-size", "1");
setMaxValue(parser, "cnv-window-size", "10000");
setDefaultValue(parser, "cnv-window-size", options.cnvWindowSize);
hideOption(parser, "cws");
addSection(parser, "Other options");
addOption(parser, ArgParseOption("lf", "log-file", "Write log to FILE.", ArgParseArgument::STRING));
addOption(parser, ArgParseOption("v", "verbose", "Enable verbose output."));
addOption(parser, ArgParseOption("vv", "very-verbose", "Enable very verbose output."));
addOption(parser, ArgParseOption("q", "quiet", "Set verbosity to a minimum."));
addTextSection(parser, "Examples");
addListItem(parser,
"\\fBsnp_store\\fP \\fB-mc\\fP \\fB2\\fP \\fB-it\\fP \\fB2\\fP \\fBexampleGenome.fa\\fP \\fBexampleReads.gff\\fP \\fB-o\\fP \\fBexampleSNPs.vcf\\fP \\fB-id\\fP \\fBexampleIndels.gff\\fP",
"Call SNPs and indels of a low-coverage example (minimum coverage and indel threshold were reduced to 2).");
addListItem(parser,
"\\fBsnp_store\\fP \\fB-re\\fP \\fB-mc\\fP \\fB2\\fP \\fB-it\\fP \\fB2\\fP \\fBexampleGenome.fa\\fP \\fBexampleReads.gff\\fP \\fB-o\\fP \\fBexampleSNPs.vcf\\fP \\fB-id\\fP \\fBexampleIndels.gff\\fP",
"Computes a realignment before variant calling. Now, the two 1bp insertions should have been merged into one 2bp insertion.");
// Parse command line.
ArgumentParser::ParseResult res = parse(parser, argc, argv);
// Only extract options if the program will continue after parseCommandLine()
if (res != ArgumentParser::PARSE_OK)
return res;
// Extract option values.
// Options:
getOptionValue(options.outputSNP, parser, "output");
if (isSet(parser, "only-successful-candidates"))
options.outputFormat = 1;
options.dontClip = isSet(parser, "dont-clip");
options.keepMultiReads = isSet(parser, "multi");
options.showQualityStrings = !isSet(parser, "hide-qualities");
if (isSet(parser, "solexa-qual-offset"))
options.asciiQualOffset = 64;
getOptionValue(options.outputIndel, parser, "indel-file");
std::string tmp;
getOptionValue(tmp, parser, "method");
options.method = (tmp == "maq")? 1: 0;
getOptionValue(options.maxPile, parser, "max-pile");
options.laneSpecificMaxPile = !isSet(parser, "merged-max-pile");
getOptionValue(options.minCoverage, parser, "min-coverage");
getOptionValue(options.forceCallCount, parser, "force-call");
options.orientationAware = isSet(parser, "orientation-aware");
// getOptionValue(options.outputCNV, parser, "output-cnv");
getOptionValue(options.outputPosition, parser, "output-positions");
getOptionValue(options.inputPositionFile, parser, "input-positions");
getOptionValue(options.maxPolymerRun, parser, "max-polymer-run");
getOptionValue(options.minDifferentReadPos, parser, "diff-pos");
getOptionValue(options.excludeBorderPos, parser, "exclude-border");
options.keepSuboptimalReads = isSet(parser, "suboptimal");
options.realign = isSet(parser, "realign");
getOptionValue(options.newQualityCalibrationFactor, parser, "corrected-quality");
getOptionValue(options.windowSize, parser, "parse-window-size");
getOptionValue(options.realignAddBorder, parser, "realign-border");
// SNP Calling Options:
getOptionValue(options.minMutT, parser, "min-mutations");
getOptionValue(options.percentageT, parser, "perc-threshold");
getOptionValue(options.avgQualT, parser, "min-quality");
getOptionValue(options.theta, parser, "theta");
getOptionValue(options.hetRate, parser, "hetero-rate");
getOptionValue(options.minMapQual, parser, "min-map-quality");
options.correctedHetTable = isSet(parser, "corrected-het");
getOptionValue(options.meanAlleleFrequency, parser, "mean-alleleFreq");
getOptionValue(options.amplificationCycles, parser, "amp-cycles");
getOptionValue(options.amplificationEfficiency, parser, "amp-efficiency");
getOptionValue(options.initialN, parser, "initial-N");
options.printHetTable = isSet(parser, "print-hetTable");
getOptionValue(options.minExplainedColumn, parser, "min-explained-column");
// Indel Calling Options:
getOptionValue(options.indelCountThreshold, parser, "indel-threshold");
getOptionValue(options.indelPercentageT, parser, "indel-perc-threshold");
getOptionValue(options.indelQualityThreshold, parser, "indel-quality-thresh");
options.bothIndelStrands = isSet(parser, "both-strands-indel");
getOptionValue(options.indelWindow, parser, "indel-window");
getOptionValue(options.indelDepthMinOverlap, parser, "exclude-border-indel");
getOptionValue(options.cnvWindowSize, parser, "cnv-window-size");
// Other Options:
getOptionValue(options.outputLog, parser, "log-file");
if (isSet(parser, "verbose"))
options._debugLevel = max(options._debugLevel, 1);
if (isSet(parser, "very-verbose"))
options._debugLevel = max(options._debugLevel, 2);
getArgumentValue(options.genomeFName, parser, 0);
unsigned countFiles = getArgumentValueCount(parser, 1);
if (countFiles == 0)
{
cerr << "No mapping files specified." << endl;
return ArgumentParser::PARSE_ERROR;
}
resize(options.readFNames, countFiles);
for (unsigned i = 0; i < countFiles; ++i)
{
getArgumentValue(options.readFNames[i], parser, 1, i);
// Get lower case of the output file name. File endings are accepted in both upper and lower case.
CharString tmp = options.readFNames[i];
toLower(tmp);
unsigned format = 0;
if (endsWith(tmp, ".gff"))
format = 0;
else if (endsWith(tmp, ".sam"))
format = 1;
else if (endsWith(tmp, ".bam"))
format = 2;
if (i == 0)
{
options.inputFormat = format;
}
else
{
if (options.inputFormat != format)
{
cerr << "All mapping files must have the same format." << endl;
return ArgumentParser::PARSE_ERROR;
}
}
}
// some additional option checking:
//if(options.inputFormat == 1 && (!options.qualityFile || (length(qualityFNames)!=length(options.readFNames))))
//{
// cerr << "If mapped read file is in Eland format, a .qual or .fastq file containing read qualities needs to be specified." << endl << endl;
// return 0;
//}
if(options.inputPositionFile == "" && options.outputPosition != "")
{
cerr << "Position analysis output specified, but no position file given." << endl << endl;
return ArgumentParser::PARSE_ERROR;
}
if((options.realign && options.windowSize > 50000) || options.windowSize > 1000000)
options.windowSize = 10000;
if (options.outputLog != "")
writeLogFile(argc, argv, options);
if(options.runID == "")
{
::std::string tempStr = toCString(options.readFNames[0]);
size_t lastPos = tempStr.find_last_of("/\\");
if (lastPos == tempStr.npos)
lastPos = 0;
else
++lastPos;
options.runID = tempStr.substr(lastPos);
}
return ArgumentParser::PARSE_OK;
}
int main(int argc, const char *argv[])
{
ArgumentParser parser;
SNPCallingOptions<> options;
ArgumentParser::ParseResult res = parseCommandLine(options, argc, argv);
// If parsing was not successful then exit with code 1 if there were errors.
// Otherwise, exit with code 0 (e.g. help was printed).
if (res != ArgumentParser::PARSE_OK)
return res == ArgumentParser::PARSE_ERROR;
for(int arg = 0; arg < argc; ++arg) {
options.programCall << argv[arg] << " ";
}
//////////////////////////////////////////////////////////////////////////////
// check for variants
int result = detectSNPs(options);
if (result > 0)
{
cerr << "ERROR: Something went wrong. Try 'snpStore --help' for more information." << endl << endl;
return 0;
}
return result;
}
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