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/*==========================================================================
SeqAn - The Library for Sequence Analysis
https://www.seqan.de
============================================================================
Copyright (C) 2007
This library 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 option) any later version.
This library 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.
============================================================================
$Id$
==========================================================================*/
#ifndef SEQAN_HEADER_OVERLAP_MODULE_H
#define SEQAN_HEADER_OVERLAP_MODULE_H
// #define DEBUG_OVERLAP_MODULE
#include <seqan/bam_io.h>
#include <seqan/store.h>
namespace seqan2
{
// Assign ContigId of children to parents
template<typename TSpec, typename TConfig>
inline void
adjustParentEntries(FragmentStore<TSpec, TConfig> & fragStore)
{
// Iterate over all parents and assign contigId of first child
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename Iterator<FragmentStore<TSpec, TConfig>, AnnotationTree<> >::Type TIter;
TIter it = begin(fragStore, AnnotationTree<>());
// Go down from root to the first parent
goDown(it);
do
{ // Iterate over all parents
if (getAnnotation(it).contigId == TAnnotationStoreElement::INVALID_ID)
{
getAnnotation(it).contigId = getAnnotation(nodeDown(it)).contigId; // If no child exists: nothing happens
getAnnotation(it).beginPos = TAnnotationStoreElement::INVALID_POS;
getAnnotation(it).endPos = TAnnotationStoreElement::INVALID_POS;
}
} while (goRight(it));
}
//////////////////////////////////////////////////////////////////////////////
// getIdsFroRead
//////////////////////////////////////////////////////////////////////////////
template<typename TAnnoIds, typename TSpec, typename TConfig, typename TIntervalTree, typename TIntervals>
inline void
getIdsForRead(TAnnoIds & ids, FragmentStore<TSpec, TConfig> & fragStore, TIntervalTree & intervalTree, TIntervals & alignIntervals, unsigned offsetInterval)
{
typedef typename FragmentStore<TSpec, TConfig>::TContigPos TContigPos;
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename TAnnotationStoreElement::TId TId;
//typedef typename Value<TIntervals>::Type TInterval;
typedef String<TId> TResult;
typedef typename Iterator<TIntervals >::Type TIntervalIter;
typedef typename Iterator<StringSet<TResult > >::Type TResultIter;
//typedef typename Iterator<TResult >::Type TIdIter;
static const TId INVALID_ID = TAnnotationStoreElement::INVALID_ID;
resize(ids, length(alignIntervals));
TIntervalIter itI = begin(alignIntervals);
TIntervalIter itIEnd = end(alignIntervals);
TResultIter itR = begin(ids);
// TResultIter itREnd = end(ids);
// search mapped annotations for each interval of the aligned read and store them in the corresponding list 'ids'
for ( ; itI != itIEnd; goNext(itI), goNext(itR))
{
findIntervalsForInterval(value(itR), intervalTree, getValue(itI), offsetInterval);
}
// check for each aligment-interval, if the inner interval-borders fit to the borders of the annotation id:
itI = begin(alignIntervals);
itR = begin(ids);
TId currentId;
TContigPos beginPos;
TContigPos endPos;
for ( ; itI != itIEnd; goNext(itI), goNext(itR))
{
for (unsigned i = 0; i < length(*itR); ++i)
{
currentId = getValue(*itR, i);
beginPos = getValue(fragStore.annotationStore, currentId).beginPos;
endPos = getValue(fragStore.annotationStore, currentId).endPos;
if (beginPos > endPos)
{
beginPos = endPos;
endPos = beginPos;
}
// begin of read
if (itR == begin(ids) && length(ids) > 1)
{
if (static_cast<TContigPos>(getValue(itI).i2 + offsetInterval) < endPos) // if the borders don't fit: delete annotation-id
{
erase(value(itR), i);
--i;
}
}
// end of read
else if (position(itR, ids) == endPosition(ids) - 1 && length(ids) > 1u)
{
if (static_cast<TContigPos>(getValue(itI).i1 - offsetInterval) > beginPos)
{
erase(value(itR), i);
--i;
}
}
// in the middle of the read
else if (length(ids) > 2)
{
if (static_cast<TContigPos>(getValue(itI).i2 + offsetInterval) < endPos)
{
erase(value(itR), i);
--i;
}
else if (static_cast<TContigPos>(getValue(itI).i1 - offsetInterval) > beginPos)
{
erase(value(itR), i);
--i;
}
}
}
if (empty(*itR) ) // if aligment-interval doesn't fit to any annotation, append INVALID_ID to mark this
appendValue(value(itR), INVALID_ID, Generous());
}
}
//////////////////////////////////////////////////////////////////////////////
////// ReadAnnoStoreELement
//////////////////////////////////////////////////////////////////////////////
template <typename TId>
struct ReadAnnoStoreElement
{
typedef StringSet<String<TId> > TAnnoIds;
TAnnoIds annoIds;
String<TId> parentIds; // not only for exon-annotations -> more than one parentId possible, only if whole read mapped in parent
TId contigId;
};
//////////////////////////////////////////////////////////////////////////////
////// assign Ids to ReadAnnoStore
//////////////////////////////////////////////////////////////////////////////
template<typename TReadAnnoStore, typename TSpec, typename TConfig, typename TId, typename TAnnoIds>
inline void
assignToReadAnnoStore(TReadAnnoStore &readAnnoStore, FragmentStore<TSpec, TConfig> & fragStore, TId readId, TAnnoIds &annoIds)
{
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename Iterator<TAnnoIds>::Type TAnnoIdsIter;
typedef typename Value<TAnnoIds>::Type TIds;
typedef typename Iterator<TIds>::Type TIdsIter;
static const TId INVALID_ID = TAnnotationStoreElement::INVALID_ID;
// assign annotationIds:
value(readAnnoStore, readId).annoIds = annoIds;
// assign coresponding parentIds:
clear (value(readAnnoStore, readId).parentIds);
if(!empty(annoIds))
{
TIdsIter itId = begin(front(annoIds));
TIdsIter itIdEnd = end(front(annoIds));
for ( ; itId != itIdEnd; goNext(itId)) // read maps in gene, if all intervals map in gene: at least one exon of the gene has to occur in the id-list of the first interval
if (getValue(itId) != INVALID_ID && !isElement_unsorted(getValue(fragStore.annotationStore, getValue(itId)).parentId, getValue(readAnnoStore, readId).parentIds))
appendValue(value(readAnnoStore, readId).parentIds, getValue(fragStore.annotationStore, getValue(itId)).parentId, Generous() );
if (!empty(getValue(readAnnoStore, readId).parentIds))
{
TAnnoIdsIter itA = begin(annoIds);
TAnnoIdsIter itAEnd = end(annoIds);
goNext(itA);
for ( ; itA != itAEnd; goNext(itA)) // not only for exon-annotations -> more than one parentId possible
{
itId = begin(*itA); // for each interval of read:
itIdEnd = end(*itA);
for (unsigned i = 0; i < length(getValue(readAnnoStore, readId).parentIds); ++i) // check if at least one child of the parentId occurs
{
for ( ; itId != itIdEnd; goNext(itId))
{
if (getValue(itId) != INVALID_ID && getValue(fragStore.annotationStore, getValue(itId)).parentId == getValue(getValue(readAnnoStore, readId).parentIds, i) )
break;
}
if (itId == itIdEnd) // if not, delete parentId
{
erase(value(readAnnoStore, readId).parentIds, i);
--i;
}
}
}
}
}
}
//////////////////////////////////////////////////////////////////////////////
////// buildTupleCountStore
//////////////////////////////////////////////////////////////////////////////
template <typename TId>
struct TupleCountStoreElement
{
typedef String<TId> TTuple;
typedef String<TTuple > TTupleList;
typedef String<unsigned> TTupleCounts;
typedef String<double> TTupleNorm;
TTupleList readConnections;
TTupleCounts readConnectionCounts;
TTupleNorm readConnectionNorm;
TTupleList matePairConnections;
TTupleCounts matePairConnectionCounts;
TTupleNorm matePairConnectionNorm;
};
//////////////////////////////////////////////////////////////////////////////
template<typename TTupleCountStore, typename TSpec, typename TConfig, typename TReadAnnoStore>
inline void
buildTupleCountStore(TTupleCountStore & tupleCountStore,
FragmentStore<TSpec, TConfig> & fragStore,
TReadAnnoStore & readAnnoStore,
unsigned n,
bool exact_nTuple)
{
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename FragmentStore<TSpec, TConfig>::TContigPos TPos;
typedef typename FragmentStore<TSpec, TConfig>::TReadStore TReadStore;
typedef typename Value<TReadStore>::Type TReadStoreElement;
typedef typename TReadStoreElement::TId TReadId;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename TAnnotationStoreElement::TId TId;
typedef typename Iterator<TReadAnnoStore>::Type TReadIter;
typedef typename Value<TReadAnnoStore>::Type TReadAnnoStoreElement;
typedef typename TReadAnnoStoreElement::TAnnoIds TAnnoIds;
typedef typename Iterator<TAnnoIds>::Type TAnnoIdsIter;
typedef typename Value<TAnnoIds>::Type TIds;
typedef typename Iterator<TIds>::Type TIdsIter;
static const TReadId INVALID_READ_ID = TReadStoreElement::INVALID_ID;
static const TId INVALID_ANNO_ID = TAnnotationStoreElement::INVALID_ID;
resize(tupleCountStore, length(fragStore.annotationStore));
bool validMate;
TReadIter itRead = begin(readAnnoStore);
TReadIter itReadEnd = end(readAnnoStore);
TIdsIter itP;
TIdsIter itPEnd;
TAnnoIds annoIds;
TAnnoIds tupleSet;
TReadId readId;
TReadId matePairId;
TReadId secReadId;
TAnnoIds secTupleSet;
TId firstAnnoId1;
TId firstAnnoId2;
TAnnoIdsIter itTuple;
TAnnoIdsIter itTupleEnd;
TAnnoIdsIter itSecTuple;
TAnnoIdsIter itSecTupleEnd;
TAnnoIdsIter itAnnoIds;
TAnnoIdsIter itAnnoIdsEnd;
TIds matePairTuple;
TPos beginPos1;
TPos endPos1;
TPos beginPos2;
TPos endPos2;
unsigned pos;
for ( ; itRead != itReadEnd; goNext(itRead))
{
if (!empty(getValue(itRead).parentIds) )
{
itP = begin(getValue(itRead).parentIds);
itPEnd = end(getValue(itRead).parentIds);
for ( ; itP != itPEnd; goNext(itP) )
{
validMate = false;
// create list of all possible tuples for current read:
annoIds = getValue(itRead).annoIds;
clear(tupleSet);
// create all Tuple of length n:
if (exact_nTuple && n <= length(annoIds)) create_nTuple(tupleSet, fragStore, annoIds, getValue(itP), n);
// create all max-Tuple (whole read) for current parentId:
else if (!exact_nTuple && n == 0) create_nTuple(tupleSet, fragStore, annoIds, getValue(itP), length(annoIds));
// create all tuple >= n for current parentId:
else if (!exact_nTuple) create_Tuple(tupleSet, fragStore, annoIds, getValue(itP), n);
if (!empty(tupleSet))
{
// create if necessary list of all possible tuples for second matepair-read:
readId = position(itRead, readAnnoStore);
matePairId = getValue(fragStore.readStore, readId).matePairId;
clear(secTupleSet);
if (matePairId != INVALID_READ_ID)
{
if(getValue(getValue(fragStore.matePairStore, matePairId).readId, 0) == readId)
secReadId = getValue(getValue(fragStore.matePairStore, matePairId).readId, 1);
else
secReadId = getValue(getValue(fragStore.matePairStore, matePairId).readId, 0);
if ( secReadId != INVALID_READ_ID )
{
//if (!empty(getValue(readAnnoStore, secReadId).annoIds))
if ( isElement_unsorted(getValue(itP), getValue(readAnnoStore, secReadId).parentIds) ) // p in parents of matepair? -> annoIds is not empty
{
validMate = true;
annoIds = getValue(readAnnoStore, secReadId).annoIds;
firstAnnoId1 = front(front(tupleSet)); // ids necessary to check positions in aligment
firstAnnoId2 = front(front(annoIds)); // can't be INVALID_ID, because parents was checked
// check if current read-position is smaller than the position of the second read -> tuple are ordered by position
if ( (getValue(fragStore.annotationStore, firstAnnoId1).beginPos <=
getValue(fragStore.annotationStore,firstAnnoId1).endPos &&
getValue(fragStore.annotationStore, firstAnnoId1).beginPos <
getValue(fragStore.annotationStore, firstAnnoId2).endPos) ||
(getValue(fragStore.annotationStore, firstAnnoId1).beginPos >
getValue(fragStore.annotationStore, firstAnnoId1).endPos &&
getValue(fragStore.annotationStore, firstAnnoId1).endPos <
getValue(fragStore.annotationStore, firstAnnoId2).beginPos) )
{
if (exact_nTuple && n <= length(annoIds)) create_nTuple(secTupleSet, fragStore, annoIds, getValue(itP), n);
else if (!exact_nTuple && n == 0) create_nTuple(secTupleSet, fragStore, annoIds, getValue(itP), length(annoIds));
else if (!exact_nTuple) create_Tuple(secTupleSet, fragStore, annoIds, getValue(itP), n);
}
}
}
}
else validMate = true;
// access to tupleCountStore for all tuple of current read:
if (validMate)
{
itTuple = begin(tupleSet);
itTupleEnd = end(tupleSet);
for ( ; itTuple != itTupleEnd; goNext(itTuple))
{
firstAnnoId1 = front(*itTuple);
erase(value(itTuple), 0); // first id is not stored; is know by position in tupleCountStore
// readConnections:
if (!empty(*itTuple))
{
if (searchValue(pos, *itTuple, getValue(tupleCountStore, firstAnnoId1).readConnections))
++value(value(tupleCountStore, firstAnnoId1).readConnectionCounts, pos);
else
{
if (pos != endPosition(getValue(tupleCountStore, firstAnnoId1).readConnections) )
{
resizeSpace(value(tupleCountStore, firstAnnoId1).readConnections, 1, pos, pos, Generous());
assignValue(value(tupleCountStore, firstAnnoId1).readConnections, pos, *itTuple);
insertValue(value(tupleCountStore, firstAnnoId1).readConnectionCounts, pos, 1, Generous());
}
else
{
appendValue(value(tupleCountStore, firstAnnoId1).readConnections, *itTuple, Generous());
appendValue(value(tupleCountStore, firstAnnoId1).readConnectionCounts, 1, Generous());
}
}
}
// matePairConnections:
if (!empty(secTupleSet))
{
itSecTuple = begin(secTupleSet);
itSecTupleEnd = end(secTupleSet);
for ( ; itSecTuple != itSecTupleEnd; goNext(itSecTuple) )
{
matePairTuple = *itTuple;
// INVALID_ID: sign for connection by matepair (apart from that, there are no INVALID_IDs in the list)
appendValue(matePairTuple, INVALID_ANNO_ID, Generous());
if (!empty(*itTuple) && back(*itTuple) == front(*itSecTuple)) // no id 2x allowed
{
if (exact_nTuple == 0 && n == 0) erase(value(itSecTuple), 0);
else continue; // tupel would be created double or tupel wouldn't have the length n anymore
}
append(matePairTuple, *itSecTuple, Generous());
if (empty(*itTuple))
{
beginPos1 = getValue(fragStore.annotationStore, firstAnnoId1).beginPos;
endPos1 = getValue(fragStore.annotationStore, firstAnnoId1).endPos;
}
else
{
beginPos1 = getValue(fragStore.annotationStore, back(*itTuple)).beginPos;
endPos1 = getValue(fragStore.annotationStore, back(*itTuple)).endPos;
}
// begin position of first annotation in tuple of second read
beginPos2 = getValue(fragStore.annotationStore, front(*itSecTuple)).beginPos;
endPos2 = getValue(fragStore.annotationStore, front(*itSecTuple)).endPos;
if ( (beginPos1 <= endPos1 && endPos1 < beginPos2) || // no overlapping annotations allowed
(endPos1 < beginPos1 && beginPos1 < endPos2) )
{
if (searchValue(pos, matePairTuple, getValue(tupleCountStore, firstAnnoId1).matePairConnections))
++value(value(tupleCountStore, firstAnnoId1).matePairConnectionCounts, pos);
else
{
if (pos != endPosition(getValue(tupleCountStore, firstAnnoId1).matePairConnections) )
{
resizeSpace(value(tupleCountStore, firstAnnoId1).matePairConnections, 1, pos, pos, Generous());
assignValue(value(tupleCountStore, firstAnnoId1).matePairConnections, pos, matePairTuple);
insertValue(value(tupleCountStore, firstAnnoId1).matePairConnectionCounts, pos, 1, Generous());
}
else
{
appendValue(value(tupleCountStore, firstAnnoId1).matePairConnections, matePairTuple, Generous());
appendValue(value(tupleCountStore, firstAnnoId1).matePairConnectionCounts, 1, Generous());
}
}
}
}
}
}
}
}
}
}
}
}
//////////////////////////////////////////////////////////////////////////////
////// buildAnnoCountStoreg++ -I../seqan/projects/library/ -D_LARGEFILE_SOURCE -D_FILE_OFFSET_BITS=64 -O3 -pedantic -lrt main.cpp -o main
//////////////////////////////////////////////////////////////////////////////
template<typename TAnnoCountStore, typename TSpec, typename TConfig, typename TReadAnnoStore>
inline void
buildAnnoCountStore(TAnnoCountStore & annoCountStore, FragmentStore<TSpec, TConfig> & fragStore, TReadAnnoStore & readAnnoStore)
{
typedef typename Iterator<TReadAnnoStore>::Type TReadIter;
typedef typename FragmentStore<TSpec, TConfig>::TReadStore TReadStore;
typedef typename Value<TReadStore>::Type TReadStoreElement;
typedef typename Value<TReadAnnoStore>::Type TReadAnnoStoreElement;
typedef typename TReadAnnoStoreElement::TAnnoIds TAnnoIds;
typedef typename Iterator<TAnnoIds>::Type TAnnoIdsIter;
typedef typename Value<TAnnoIds>::Type TIds;
typedef typename Iterator<TIds>::Type TIdsIter;
typedef typename Value<TIds>::Type TId;
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
static const TId INVALID_READ_ID = TReadStoreElement::INVALID_ID;
static const TId INVALID_ANNO_ID = TAnnotationStoreElement::INVALID_ID;
resize(annoCountStore, length(fragStore.annotationStore), 0);
TReadIter itRead = begin(readAnnoStore);
TReadIter itReadEnd = end(readAnnoStore);
TId readId;
TId matePairId;
TId secReadId = 0;
TIds interSecIds;
TIdsIter itP;
TIdsIter itPEnd;
TAnnoIdsIter itAnnoIds;
TAnnoIdsIter itAnnoIdsEnd;
TIdsIter itId;
TIdsIter itIdEnd;
// increment for each read respective to its mapped ids the count in the annoCountStore
for ( ; itRead != itReadEnd; goNext(itRead))
{
if (!empty(getValue(itRead).annoIds) )
{
readId = position(itRead, readAnnoStore);
matePairId = getValue(fragStore.readStore, readId).matePairId;
if (matePairId != INVALID_READ_ID)
{
if (getValue(getValue(fragStore.matePairStore, matePairId).readId, 0) == readId)
secReadId = getValue(getValue(fragStore.matePairStore, matePairId).readId, 1);
else
secReadId = getValue(getValue(fragStore.matePairStore, matePairId).readId, 0);
}
// for each parentId: we just want to count annotations, in which the read mapped
if (!empty(getValue(itRead).parentIds))
{
itP = begin(getValue(itRead).parentIds);
itPEnd = end(getValue(itRead).parentIds);
for (; itP != itPEnd; goNext(itP) )
{
// check mate-read to prevent double counts
if (matePairId != INVALID_READ_ID)
{
if (!isElement_unsorted(getValue(itP), getValue(readAnnoStore, secReadId).parentIds) )
continue; // if matepair read doesn't map in same parentId: no count (go to next parentId)
// count annotations, which occur in both reads, shouldn't be increment for the read with the bigger readId
if (secReadId < readId )
{
clear(interSecIds);
// just check the periphery annotations
// if the end of the current read mapped in a same annotation as the start of the second read:
if ( interSec(interSecIds, back(getValue(itRead).annoIds), front(getValue(readAnnoStore, secReadId).annoIds)) )
{
for (unsigned i = 0; i < length(interSecIds); ++i)
{
if (getValue(fragStore.annotationStore, getValue(interSecIds, i) ).parentId == getValue(itP))
--value(annoCountStore, getValue(interSecIds, i));
// decrement the corresponding count
}
}
// or if the start of the current read mapped in a same annotation as the end of the second read:
else if ( interSec(interSecIds, front(getValue(itRead).annoIds),
back(getValue(readAnnoStore, secReadId).annoIds)) )
{
for (unsigned i = 0; i < length(interSecIds); ++i)
{
if (getValue(fragStore.annotationStore, getValue(interSecIds, i) ).parentId == getValue(itP))
--value(annoCountStore, getValue(interSecIds, i));
}
}
if (getValue(itP) != INVALID_ANNO_ID) --value(annoCountStore, getValue(itP));
}
}
// count for all annoIds
itAnnoIds = begin(getValue(itRead).annoIds);
itAnnoIdsEnd = end(getValue(itRead).annoIds);
for ( ; itAnnoIds != itAnnoIdsEnd; goNext(itAnnoIds))
{
itId = begin(*itAnnoIds);
itIdEnd = end(*itAnnoIds);
for ( ; itId != itIdEnd; goNext(itId))
if (getValue(itId) != INVALID_ANNO_ID && getValue(fragStore.annotationStore, getValue(itId)).parentId == getValue(itP) )
++value(annoCountStore, getValue(itId));
}
// count for parentIds (already selected)
if (getValue(itP) != INVALID_ANNO_ID) ++value(annoCountStore, getValue(itP));
}
}
}
}
}
//////////////////////////////////////////////////////////////////////////////
////// Overlap Module
//////////////////////////////////////////////////////////////////////////////
template<typename TReadAnnoStore, typename TAnnoCountStore, typename TTupleCountStore, typename TSpec, typename TConfig>
inline void
getResults(TReadAnnoStore & readAnnoStore,
TAnnoCountStore & annoCountStore,
TTupleCountStore & tupleCountStore,
FragmentStore<TSpec, TConfig> & fragStore,
unsigned tupelSize,
bool exact_nTuple,
unsigned offsetInterval,
unsigned thresholdGaps,
bool unknownO)
{
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename TAnnotationStoreElement::TId TId;
typedef typename FragmentStore<TSpec, TConfig>::TIntervalTreeStore TIntervalTreeStore;
typedef typename Iterator<TIntervalTreeStore>::Type TIntervalTree;
typedef typename Value<TReadAnnoStore>::Type TReadAnnoStoreElement;
typedef typename TReadAnnoStoreElement::TAnnoIds TAnnoIds;
typedef typename FragmentStore<TSpec, TConfig>::TAlignedReadStore TAlignedReadStore;
typedef typename Position<TAlignedReadStore>::Type TAlignPos;
typedef String<AlignIntervalsStoreElement<> > TAlignIntervalsStore;
typedef typename Iterator<TAlignIntervalsStore>::Type TAlignIntervalsStoreIter;
resize(readAnnoStore, length(fragStore.readStore));
TIntervalTree intervalTree;
// extract intervals from alignedReadStore and store them in AlignIntervalsStore:
TAlignIntervalsStore alignIntervalsStore;
buildAlignIntervalsStore(alignIntervalsStore, fragStore, thresholdGaps);
if (!empty(alignIntervalsStore))
{
TAlignPos alignPos;
TId contigId;
TId readId;
TAnnoIds ids;
TAlignIntervalsStoreIter it = begin(alignIntervalsStore);
TAlignIntervalsStoreIter itEnd = end(alignIntervalsStore);
// for each item in alignIntervalsStore:
for ( ; it != itEnd; goNext(it))
{
// get ids from alignedReadStore (same position as in alignIntervalsStore):
alignPos = position(it, alignIntervalsStore);
contigId = getValue(fragStore.alignedReadStore, alignPos).contigId;
readId = getValue(fragStore.alignedReadStore, alignPos).readId;
// get respective intervalTree
if (unknownO || getValue(fragStore.alignedReadStore, alignPos).beginPos <= getValue(fragStore.alignedReadStore, alignPos).endPos)
intervalTree = begin(fragStore.intervalTreeStore_F, Standard()) + contigId; //getValue(fragStore.intervalTreeStore_F, contigId);
else
intervalTree = begin(fragStore.intervalTreeStore_R, Standard()) + contigId; //getValue(fragStore.intervalTreeStore_R, contigId);
// get annotationStore-Ids for these intervals:
clear(ids);
if ((*intervalTree).interval_counter != 0)
getIdsForRead(ids, fragStore, *intervalTree, getValue(it).intervals, offsetInterval);
// assign Ids from mapped annotations to readAnnoStore:
value(readAnnoStore, readId).contigId = contigId;
assignToReadAnnoStore(readAnnoStore, fragStore, readId, ids);
}
}
buildAnnoCountStore(annoCountStore, fragStore, readAnnoStore);
buildTupleCountStore(tupleCountStore, fragStore, readAnnoStore, tupelSize, exact_nTuple);
}
//////////////////////////////////////////////////////////////////////////////
/// get normalized values for annotations And get Map for Gene orientations (necessary for annotation Output)
//////////////////////////////////////////////////////////////////////////////
template<typename TAnnoNormStore, typename TMapO, typename TAnnoCountStore, typename TSpec, typename TConfig>
inline void
normalizeAnnoCounts(TAnnoNormStore &annoNormStore, TMapO &mapO, TAnnoCountStore &annoCountStore, FragmentStore<TSpec, TConfig> &fragStore)
{
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename TAnnotationStoreElement::TId TId;
typedef typename FragmentStore<TSpec, TConfig>::TContigPos TPos;
typedef typename FragmentStore<TSpec, TConfig>::TReadStore TReadStore;
typedef typename Size<TReadStore>::Type TReadStoreSize;
typedef typename Iterator<TAnnotationStore>::Type TAnnoIter;
typedef typename Iterator<TAnnoCountStore>::Type TCountIter;
typedef typename Iterator<TAnnoNormStore>::Type TNormIter;
typedef typename Size<TPos>::Type TSize;
typedef String<TSize> TChildrenLengths;
typedef typename Iterator<String<TSize> >::Type TLengthIter;
typedef Pair<TId, TChildrenLengths> TPair;
typedef typename Value<TMapO>::Type TPairO;
typedef Map<TPair> TMap;
typedef typename Iterator<TMap>::Type TMapIter;
static const TId INVALID_ID = TAnnotationStoreElement::INVALID_ID;
static const TPos INVALID_POS = TAnnotationStoreElement::INVALID_POS;
resize(annoNormStore, length(annoCountStore), 0);
TReadStoreSize readNo = length(fragStore.readStore) - length(fragStore.matePairStore);
TMap map;
clear(map);
clear(mapO);
if(!empty(fragStore.annotationStore))
{
TAnnoIter itA = begin(fragStore.annotationStore);
TAnnoIter itAEnd = end(fragStore.annotationStore);
TCountIter itC = begin(annoCountStore);
TNormIter itN = begin(annoNormStore);
TChildrenLengths childrenLengths;
TPair pair;
TPairO pairO;
TSize length;
for ( ; itA != itAEnd; goNext(itA), goNext(itC), goNext(itN) )
{
if (getValue(itA).beginPos == INVALID_POS && getValue(itA).parentId == INVALID_ID) // make entry for each gene/parent in map:
{
clear(childrenLengths);
pair.i1 = position(itA, fragStore.annotationStore);
pair.i2 = childrenLengths;
insert(map, pair); // for lengths
pairO.i1 = position(itA, fragStore.annotationStore);
pairO.i2 = 0;
insert(mapO, pairO); // for orientation
}
else if (getValue(itA).beginPos != INVALID_POS) // for each exon/child:
{
if (getValue(itA).beginPos <= getValue(itA).endPos)
length = getValue(itA).endPos - getValue(itA).beginPos;
else
length = getValue(itA).beginPos - getValue(itA).endPos;
value(itN) = ((double)1000000000 * (double)getValue(itC))/((double)readNo * (double)length); // calculate normalized expression-value
if (getValue(itA).parentId != INVALID_ID) // append length to gene/parent lengths
{
appendValue(mapValue(map, getValue(itA).parentId), length, Generous());
if (getValue(itA).beginPos > getValue(itA).endPos)
mapValue(mapO, getValue(itA).parentId) = 1;
}
}
}
}
if (!empty(map))
{
TMapIter itM = begin(map);
TMapIter itMEnd = end(map);
TSize length;
TLengthIter itL;
TLengthIter itLEnd;
for ( ; itM != itMEnd; goNext(itM)) // calculate normalized gene/parent expression-values
{
length = 0;
itL = begin(value(itM).i2);
itLEnd = end(value(itM).i2);
for ( ; itL != itLEnd; goNext(itL))
length += getValue(itL);
value(annoNormStore, value(itM).i1) = ((double)1000000000 * (double)getValue(annoCountStore, value(itM).i1) )/((double)readNo * (double)length);
}
}
}
//////////////////////////////////////////////////////////////////////////////
/// get normalized values for tuple
//////////////////////////////////////////////////////////////////////////////
template<typename TTupleCountStore, typename TSpec, typename TConfig>
inline void
normalizeTupleCounts(TTupleCountStore &tupleCountStore, FragmentStore<TSpec, TConfig> &fragStore)
{
typedef typename FragmentStore<TSpec, TConfig>::TAnnotationStore TAnnotationStore;
typedef typename Value<TAnnotationStore>::Type TAnnotationStoreElement;
typedef typename TAnnotationStoreElement::TId TId;
typedef typename Value<TTupleCountStore>::Type TTupleCountStoreElement;
typedef typename TTupleCountStoreElement::TTupleList TTupleList;
typedef typename TTupleCountStoreElement::TTupleCounts TTupleCounts;
typedef typename TTupleCountStoreElement::TTupleNorm TTupleNorm;
typedef typename TTupleCountStoreElement::TTuple TTuple;
typedef typename Iterator<TTupleCountStore>::Type TStoreIter;
typedef typename Iterator<TTupleList>::Type TTupleListIter;
typedef typename Iterator<TTupleCounts>::Type TCountIter;
typedef typename Iterator<TTupleNorm>::Type TNormIter;
typedef typename Iterator<TTuple>::Type TTupleIter;
typedef typename FragmentStore<TSpec, TConfig>::TContigPos TPos;
typedef typename Size<TPos>::Type TSize;
typedef typename FragmentStore<TSpec, TConfig>::TReadStore TReadStore;
typedef typename Size<TReadStore>::Type TReadStoreSize;
static const TId INVALID_ID = TAnnotationStoreElement::INVALID_ID;
TReadStoreSize readNo = length(fragStore.readStore) - length(fragStore.matePairStore);
if (!empty(tupleCountStore))
{
TStoreIter itS = begin(tupleCountStore);
TStoreIter itSEnd = end(tupleCountStore);
TTupleListIter itT;
TTupleListIter itTEnd;
TCountIter itC;
TNormIter itN;
TSize tupleLength;
TTupleIter itId;
TTupleIter itIdEnd;
for ( ; itS != itSEnd; goNext(itS))
{
// readConnections:
resize(value(itS).readConnectionNorm, length(getValue(itS).readConnections));
if (!empty(getValue(itS).readConnections))
{
itT = begin(itS->readConnections);
itTEnd = end(itS->readConnections);
itC = begin(itS->readConnectionCounts);
itN = begin(itS->readConnectionNorm);
for ( ; itT != itTEnd; goNext(itT), goNext(itC), goNext(itN))
{
tupleLength = 0;
itId = begin(*itT);
itIdEnd = end(*itT);
for ( ; itId != itIdEnd; goNext(itId))
{
if (getValue(fragStore.annotationStore, *itId).beginPos <= getValue(fragStore.annotationStore, *itId).endPos)
tupleLength += getValue(fragStore.annotationStore, *itId).endPos - getValue(fragStore.annotationStore, *itId).beginPos;
else
tupleLength += getValue(fragStore.annotationStore, *itId).beginPos - getValue(fragStore.annotationStore, *itId).endPos;
}
value(itN) = ((double)1000000000 * (double)*itC) / ((double)readNo * (double)tupleLength);
}
}
// matePairConnections:
resize(value(itS).matePairConnectionNorm, length(itS->matePairConnections));
if (!empty(itS->matePairConnections))
{
itT = begin(itS->matePairConnections);
itTEnd = end(itS->matePairConnections);
itC = begin(itS->matePairConnectionCounts);
itN = begin(itS->matePairConnectionNorm);
for ( ; itT != itTEnd; goNext(itT), goNext(itC), goNext(itN))
{
tupleLength = 0;
itId = begin(*itT);
itIdEnd = end(*itT);
for ( ; itId != itIdEnd; goNext(itId))
{
if (*itId != INVALID_ID)
{
if (getValue(fragStore.annotationStore, *itId).beginPos <= getValue(fragStore.annotationStore, *itId).endPos)
tupleLength += getValue(fragStore.annotationStore, *itId).endPos - getValue(fragStore.annotationStore, *itId).beginPos;
else
tupleLength += getValue(fragStore.annotationStore, getValue(itId)).beginPos - getValue(fragStore.annotationStore, *itId).endPos;
}
}
value(itN) = ((double)1000000000 * (double)*itC) / ((double)readNo * (double)tupleLength);
}
}
}
}
}
//////////////////////////////////////////////////////////////////////////////
/// NGS Overlapper main function
//////////////////////////////////////////////////////////////////////////////
template<typename TOptions>
inline bool
ngsOverlapper(TOptions &options)
{
FragmentStore<> fragStore;
#ifdef DEBUG_OVERLAP_MODULE
SEQAN_PROTIMESTART(find1_time);
#endif
// build contigStore from FASTA file
#ifdef DEBUG_OVERLAP_MODULE
std::cout << "load Sam..." << std::endl;
#endif
// read aligned reads in FragmentStore from Sam files
BamFileIn inSam(toCString(options.nameSAM));
readRecords(fragStore, inSam);
#ifdef DEBUG_OVERLAP_MODULE
std::cout << " loaded " << length(fragStore.readSeqStore) << " read records" << std::endl;
#endif
#ifdef DEBUG_OVERLAP_MODULE
SEQAN_PROTIMESTART(find2_time);
#endif
// read annotations from GFF or GTF
GffFileIn inGff(toCString(options.nameGFF));
readRecords(fragStore, inGff);
adjustParentEntries(fragStore);
// create IntervalTreeStore:
#ifdef DEBUG_OVERLAP_MODULE
SEQAN_PROTIMESTART(find3_time);
#endif
createIntervalTreeStore(fragStore, options.unknownO);
#ifdef DEBUG_OVERLAP_MODULE
std::cout << "create intervalTreeStores from annotationStore took: \t" << SEQAN_PROTIMEDIFF(find3_time) << " seconds" << std::endl;
#endif
// build stores for results:
String<ReadAnnoStoreElement<unsigned> > readAnnoStore;
String<unsigned> annoCountStore;
String<TupleCountStoreElement<unsigned> > tupleCountStore;
String<TupleCountStoreElement_Fusion<unsigned> > tupleCountStore_Fusion; // additional Store, if fusion genes should be checked
// get results with additional check for transfusion genes (will be changed later additionally)
if (options.fusion == 1)
getResults_Fusion(readAnnoStore, annoCountStore, tupleCountStore, tupleCountStore_Fusion, fragStore, options.nTuple, options.exact_nTuple, options.offsetInterval, options.thresholdGaps, options.unknownO);
else // get normal results:
getResults(readAnnoStore, annoCountStore, tupleCountStore, fragStore, options.nTuple, options.exact_nTuple, options.offsetInterval, options.thresholdGaps, options.unknownO);
// normalize:
String<double> annoNormStore;
Map<Pair<unsigned, bool> > mapO;
normalizeAnnoCounts(annoNormStore, mapO, annoCountStore, fragStore);
normalizeTupleCounts(tupleCountStore, fragStore);
if (options.fusion == 1)
normalizeTupleCounts_Fusion(tupleCountStore_Fusion, fragStore);
// output:
std::fstream readOutput;
readOutput.open(toCString(options.readOutputFileName), std::ios_base::out | std::ios_base::trunc | std::ios_base::binary);
if (!readOutput.good())
std::cerr << "ERROR: Could not open output file " << options.readOutputFileName << "\n";
createReadCountGFF(readOutput, readAnnoStore, fragStore);
readOutput.close();
std::fstream annoOutput;
annoOutput.open(toCString(options.annoOutputFileName), std::ios_base::out | std::ios_base::trunc | std::ios_base::binary);
if (!annoOutput.good())
std::cerr << "ERROR: Could not open output file " << options.annoOutputFileName << "\n";
createAnnoCountGFF(annoOutput, annoCountStore, annoNormStore, fragStore, mapO);
annoOutput.close();
std::fstream tupleOutput;
tupleOutput.open(toCString(options.tupleOutputFileName), std::ios_base::out | std::ios_base::trunc | std::ios_base::binary);
if (!tupleOutput.good())
std::cerr << "ERROR: Could not open output file " << options.tupleOutputFileName << "\n";
createTupleCountGFF(tupleOutput, tupleCountStore, fragStore, options.thresholdCount, options.thresholdRPKM);
tupleOutput.close();
// additional output, if fusion genes were checked
if (options.fusion == 1)
{
std::fstream tupleOutput_Fusion;
tupleOutput_Fusion.open(toCString(options.tupleFusionOutputFileName), std::ios_base::out | std::ios_base::trunc | std::ios_base::binary);
createTupleCountGFF_Fusion(tupleOutput_Fusion, tupleCountStore_Fusion, fragStore, options.thresholdCount, options.thresholdRPKM);
tupleOutput_Fusion.close();
}
#ifdef DEBUG_OVERLAP_MODULE
std::cout << "ngsOverlapper-function took: \t" << SEQAN_PROTIMEDIFF(find1_time) << " seconds" << std::endl;
std::cout << "ngsOverlapper-function without reading Sam took: \t" << SEQAN_PROTIMEDIFF(find2_time) << " seconds" << std::endl;
std::cout << "ngsOverlapper-function and create IntervalTreeStore without reading Sam took:\t" << SEQAN_PROTIMEDIFF(find2_time) - SEQAN_PROTIMEDIFF(find3_time) << " seconds" << std::endl;
#endif
return true;
}
//////////////////////////////////////////////////////////////////////////////
}// namespace seqan2
#endif //#ifndef SEQAN_HEADER_...
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