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/*++
Module Name:
IntersectingPairedEndAligner.h
Abstract:
A paired-end aligner based on set intersections to narrow down possible candidate locations.
Authors:
Bill Bolosky, February, 2013
Environment:
User mode service.
Revision History:
--*/
#pragma once
#include "PairedEndAligner.h"
#include "BaseAligner.h"
#include "BigAlloc.h"
#include "directions.h"
#include "LandauVishkin.h"
#include "FixedSizeMap.h"
const unsigned DEFAULT_INTERSECTING_ALIGNER_MAX_HITS = 2000;
const unsigned DEFAULT_MAX_CANDIDATE_POOL_SIZE = 1000000;
class IntersectingPairedEndAligner : public PairedEndAligner
{
public:
IntersectingPairedEndAligner(
GenomeIndex *index_,
unsigned maxReadSize_,
unsigned maxHits_,
unsigned maxK_,
unsigned maxSeedsFromCommandLine_,
double seedCoverage_,
unsigned minSpacing_, // Minimum distance to allow between the two ends.
unsigned maxSpacing_, // Maximum distance to allow between the two ends.
unsigned maxBigHits_,
unsigned extraSearchDepth_,
unsigned maxCandidatePoolSize,
int maxSecondaryAlignmentsPerContig_,
BigAllocator *allocator,
bool noUkkonen_,
bool noOrderedEvaluation_,
bool noTruncation_);
static unsigned getMaxSecondaryResults(unsigned numSeedsFromCommandLine, double seedCoverage, unsigned maxReadSize, unsigned maxHits, unsigned seedLength, unsigned minSpacing, unsigned maxSpacing)
{
unsigned maxSeedsToUse;
if (0 != numSeedsFromCommandLine) {
maxSeedsToUse = numSeedsFromCommandLine;
} else {
maxSeedsToUse = (unsigned)(maxReadSize * seedCoverage / seedLength);
}
//
// The number of hits we can conceivably get is for each seed a result for every hit, times every possible pair for that hit. The possible pairs
// run from min to max distance on either side, but if they're within max merge distance then they'll be merged.
//
return NUM_DIRECTIONS * maxHits * maxSeedsToUse * (maxSpacing - minSpacing + 1 + maxMergeDistance - 1) / maxMergeDistance * 2;
}
void setLandauVishkin(
LandauVishkin<1> *landauVishkin_,
LandauVishkin<-1> *reverseLandauVishkin_)
{
landauVishkin = landauVishkin_;
reverseLandauVishkin = reverseLandauVishkin_;
}
virtual ~IntersectingPairedEndAligner();
virtual void align(
Read *read0,
Read *read1,
PairedAlignmentResult *result,
int maxEditDistanceForSecondaryResults,
int secondaryResultBufferSize,
int *nSecondaryResults,
PairedAlignmentResult *secondaryResults, // The caller passes in a buffer of secondaryResultBufferSize and it's filled in by align()
int singleSecondaryBufferSize,
int maxSecondaryResultsToReturn,
int *nSingleEndSecondaryResultsForFirstRead,
int *nSingleEndSecondaryResultsForSecondRead,
SingleAlignmentResult *singleEndSecondaryResults // Single-end secondary alignments for when the paired-end alignment didn't work properly
);
static size_t getBigAllocatorReservation(GenomeIndex * index, unsigned maxBigHitsToConsider, unsigned maxReadSize, unsigned seedLen, unsigned maxSeedsFromCommandLine,
double seedCoverage, unsigned maxEditDistanceToConsider, unsigned maxExtraSearchDepth, unsigned maxCandidatePoolSize,
int maxSecondaryAlignmentsPerContig);
void *operator new(size_t size, BigAllocator *allocator) {_ASSERT(size == sizeof(IntersectingPairedEndAligner)); return allocator->allocate(size);}
void operator delete(void *ptr, BigAllocator *allocator) {/* do nothing. Memory gets cleaned up when the allocator is deleted.*/}
void *operator new(size_t size) {return BigAlloc(size);}
void operator delete(void *ptr) {BigDealloc(ptr);}
virtual _int64 getLocationsScored() const {
return nLocationsScored;
}
private:
IntersectingPairedEndAligner() {} // This is for the counting allocator, it doesn't build a useful object
static const int NUM_SET_PAIRS = 2; // A "set pair" is read0 FORWARD + read1 RC, or read0 RC + read1 FORWARD. Again, it doesn't make sense to change this.
void allocateDynamicMemory(BigAllocator *allocator, unsigned maxReadSize, unsigned maxBigHitsToConsider, unsigned maxSeedsToUse,
unsigned maxEditDistanceToConsider, unsigned maxExtraSearchDepth, unsigned maxCandidatePoolSize,
int maxSecondaryAlignmentsPerContig);
GenomeIndex * index;
const Genome * genome;
GenomeDistance genomeSize;
unsigned maxReadSize;
unsigned maxHits;
unsigned maxBigHits;
unsigned extraSearchDepth;
unsigned maxK;
unsigned numSeedsFromCommandLine;
double seedCoverage;
static const unsigned MAX_MAX_SEEDS = 30;
unsigned minSpacing;
unsigned maxSpacing;
unsigned seedLen;
bool doesGenomeIndexHave64BitLocations;
_int64 nLocationsScored;
bool noUkkonen;
bool noOrderedEvaluation;
bool noTruncation;
static const unsigned maxMergeDistance;
//
// It's a template, because we
// have different sizes of genome locations depending on the hash table format. So, GL must be unsigned or GenomeLocation
//
template<class GL> struct HashTableLookup {
unsigned seedOffset;
_int64 nHits;
const GL * hits;
unsigned whichDisjointHitSet;
//
// We keep the hash table lookups that haven't been exhaused in a circular list.
//
HashTableLookup<GL> *nextLookupWithRemainingMembers;
HashTableLookup<GL> *prevLookupWithRemainingMembers;
//
// State for handling the binary search of a location in this lookup.
// This would ordinarily be stack local state in the binary search
// routine, but because a) we want to interleave the steps of the binary
// search in order to allow cache prefetches to have time to execute;
// and b) we don't want to do dynamic memory allocation (really at all),
// it gets stuck here.
//
int limit[2]; // The upper and lower limits of the current binary search in hits
GL maxGenomeLocationToFindThisSeed;
//
// A linked list of lookups that haven't yet completed this binary search. This is a linked
// list with no header element, so testing for emptiness needs to happen at removal time.
// It's done that way to avoid a comparison for list head that would result in a hard-to-predict
// branch.
//
HashTableLookup<GL> *nextLookupForCurrentBinarySearch;
HashTableLookup<GL> *prevLookupForCurrentBinarySearch;
_int64 currentHitForIntersection;
//
// A place for the hash table to write in singletons. We need this because when the hash table is
// built with > 4 byte genome locations, it usually doesn't store 8 bytes, so we need to
// provide the lookup function a place to write the result. Since we need one per
// lookup, it goes here.
//
GL singletonGenomeLocation[2]; // The [2] is because we need to look one before sometimes, and that allows space
};
//
// A set of seed hits, represented by the lookups that came out of the big hash table. It can be over 32 or
// 64 bit indices, but its external interface is always 64 bits (it extends on the way out if necessary).
//
class HashTableHitSet {
public:
HashTableHitSet() {}
void firstInit(unsigned maxSeeds_, unsigned maxMergeDistance_, BigAllocator *allocator, bool doesGenomeIndexHave64BitLocations_);
//
// Reset to empty state.
//
void init();
//
// Record a hash table lookup. All recording must be done before any
// calls to getNextHitLessThanOrEqualTo. A disjoint hit set is a set of hits
// that don't share any bases in the read. This is interesting because the edit
// distance of a read must be at least the number of seeds that didn't hit for
// any disjoint hit set (because there must be a difference in the read within a
// seed for it not to hit, and since the reads are disjoint there can't be a case
// where the same difference caused two seeds to miss).
//
void recordLookup(unsigned seedOffset, _int64 nHits, const unsigned *hits, bool beginsDisjointHitSet);
void recordLookup(unsigned seedOffset, _int64 nHits, const GenomeLocation *hits, bool beginsDisjointHitSet);
//
// This efficiently works through the set looking for the next hit at or below this address.
// A HashTableHitSet only allows a single iteration through its address space per call to
// init().
//
bool getNextHitLessThanOrEqualTo(GenomeLocation maxGenomeLocationToFind, GenomeLocation *actualGenomeLocationFound, unsigned *seedOffsetFound);
//
// Walk down just one step, don't binary search.
//
bool getNextLowerHit(GenomeLocation *genomeLocation, unsigned *seedOffsetFound);
//
// Find the highest genome address.
//
bool getFirstHit(GenomeLocation *genomeLocation, unsigned *seedOffsetFound);
unsigned computeBestPossibleScoreForCurrentHit();
//
// This is bit of storage that the 64 bit lookup needs in order to extend singleton hits into 64 bits, since they may be
// stored in the index in fewer.
//
GenomeLocation *getNextSingletonLocation()
{
return &lookups64[nLookupsUsed].singletonGenomeLocation[1];
}
private:
struct DisjointHitSet {
unsigned countOfExhaustedHits;
unsigned missCount;
};
int currentDisjointHitSet;
DisjointHitSet * disjointHitSets;
HashTableLookup<unsigned> * lookups32;
HashTableLookup<GenomeLocation> * lookups64;
HashTableLookup<unsigned> lookupListHead32[1];
HashTableLookup<GenomeLocation> lookupListHead64[1];
unsigned maxSeeds;
unsigned nLookupsUsed;
GenomeLocation mostRecentLocationReturned;
unsigned maxMergeDistance;
bool doesGenomeIndexHave64BitLocations;
};
HashTableHitSet * hashTableHitSets[NUM_READS_PER_PAIR][NUM_DIRECTIONS];
int countOfHashTableLookups[NUM_READS_PER_PAIR];
_int64 totalHashTableHits[NUM_READS_PER_PAIR][NUM_DIRECTIONS];
_int64 largestHashTableHit[NUM_READS_PER_PAIR][NUM_DIRECTIONS];
unsigned readWithMoreHits;
unsigned readWithFewerHits;
//
// A location that's been scored (or waiting to be scored). This is needed in order to do merging
// of close-together hits and to track potential mate pairs.
//
struct HitLocation {
GenomeLocation genomeLocation;
int genomeLocationOffset; // This is needed because we might get an offset back from scoring (because it's really scoring a range).
unsigned seedOffset;
bool isScored; // Mate pairs are sometimes not scored when they're inserted, because they
unsigned score;
unsigned maxK; // The maxK that this was scored with (we may need to rescore if we need a higher maxK and score is -1)
double matchProbability;
unsigned bestPossibleScore;
//
// We have to be careful in the case where lots of offsets in a row match well against the read (think
// about repetitive short sequences, i.e., ATTATTATTATT...). We want to merge the close ones together,
// but if the repetitive sequence extends longer than maxMerge, we don't want to just slide the window
// over the whole range and declare it all to be one. There is really no good definition for the right
// thing to do here, so instead all we do is that when we declare two candidates to be matched we
// pick one of them to be the match primary and then coalesce all matches that are within maxMatchDistance
// of the match primary. No one can match with any of the locations in the set that's beyond maxMatchDistance
// from the set primary. This means that in the case of repetitve sequences that we'll declare locations
// right next to one another not to be matches. There's really no way around this while avoiding
// matching things that are possibly much more than maxMatchDistance apart.
//
GenomeLocation genomeLocationOfNearestMatchedCandidate;
};
char *rcReadData[NUM_READS_PER_PAIR]; // the reverse complement of the data for each read
char *rcReadQuality[NUM_READS_PER_PAIR]; // the reversed quality strings for each read
unsigned readLen[NUM_READS_PER_PAIR];
Read *reads[NUM_READS_PER_PAIR][NUM_DIRECTIONS]; // These are the reads that are provided in the align call, together with their reverse complements, which are computed.
Read rcReads[NUM_READS_PER_PAIR][NUM_DIRECTIONS];
char *reversedRead[NUM_READS_PER_PAIR][NUM_DIRECTIONS]; // The reversed data for each read for forward and RC. This is used in the backwards LV
LandauVishkin<> *landauVishkin;
LandauVishkin<-1> *reverseLandauVishkin;
char rcTranslationTable[256];
unsigned nTable[256];
BYTE *seedUsed;
inline bool IsSeedUsed(_int64 indexInRead) const {
return (seedUsed[indexInRead / 8] & (1 << (indexInRead % 8))) != 0;
}
inline void SetSeedUsed(_int64 indexInRead) {
seedUsed[indexInRead / 8] |= (1 << (indexInRead % 8));
}
//
// "Local probability" means the probability that each end is correct given that the pair itself is correct.
// Consider the example where there's exactly one decent match for one read, but the other one has several
// that are all within the correct range for the first one. Then the local probability for the second read
// is lower than the first. The overall probability of an alignment then is
// pairProbability * localProbability/ allPairProbability.
//
double localBestPairProbability[NUM_READS_PER_PAIR];
void scoreLocation(
unsigned whichRead,
Direction direction,
GenomeLocation genomeLocation,
unsigned seedOffset,
unsigned scoreLimit,
unsigned *score,
double *matchProbability,
int *genomeLocationOffset // The computed offset for genomeLocation (which is needed because we scan several different possible starting locations)
);
//
// These are used to keep track of places where we should merge together candidate locations for MAPQ purposes, because they're sufficiently
// close in the genome.
//
struct MergeAnchor {
double matchProbability;
GenomeLocation locationForReadWithMoreHits;
GenomeLocation locationForReadWithFewerHits;
int pairScore;
void init(GenomeLocation locationForReadWithMoreHits_, GenomeLocation locationForReadWithFewerHits_, double matchProbability_, int pairScore_) {
locationForReadWithMoreHits = locationForReadWithMoreHits_;
locationForReadWithFewerHits = locationForReadWithFewerHits_;
matchProbability = matchProbability_;
pairScore = pairScore_;
}
//
// Returns whether this candidate is a match for this merge anchor.
//
bool doesRangeMatch(GenomeLocation newMoreHitLocation, GenomeLocation newFewerHitLocation) {
GenomeDistance deltaMore = DistanceBetweenGenomeLocations(locationForReadWithMoreHits, newMoreHitLocation);
GenomeDistance deltaFewer = DistanceBetweenGenomeLocations(locationForReadWithFewerHits, newFewerHitLocation);
return deltaMore < 50 && deltaFewer < 50;
}
//
// Returns true and sets oldMatchProbability if this should be eliminated due to a match.
//
bool checkMerge(GenomeLocation newMoreHitLocation, GenomeLocation newFewerHitLocation, double newMatchProbability, int newPairScore,
double *oldMatchProbability);
};
//
// We keep track of pairs of locations to score using two structs, one for each end. The ends for the read with fewer hits points into
// a list of structs for the end with more hits, so that we don't need one stuct for each pair, just one for each end, and also so that
// we don't need to score the mates more than once if they could be paired with more than one location from the end with fewer hits.
//
struct ScoringMateCandidate {
//
// These are kept in arrays in decreasing genome order, one for each set pair, so you can find the next largest location by just looking one
// index lower, and vice versa.
//
double matchProbability;
GenomeLocation readWithMoreHitsGenomeLocation;
unsigned bestPossibleScore;
unsigned score;
unsigned scoreLimit; // The scoreLimit with which score was computed
unsigned seedOffset;
int genomeOffset;
void init(GenomeLocation readWithMoreHitsGenomeLocation_, unsigned bestPossibleScore_, unsigned seedOffset_) {
readWithMoreHitsGenomeLocation = readWithMoreHitsGenomeLocation_;
bestPossibleScore = bestPossibleScore_;
seedOffset = seedOffset_;
score = -2;
scoreLimit = -1;
matchProbability = 0;
genomeOffset = 0;
}
};
struct ScoringCandidate {
ScoringCandidate * scoreListNext; // This is a singly-linked list
MergeAnchor * mergeAnchor;
unsigned scoringMateCandidateIndex; // Index into the array of scoring mate candidates where we should look
GenomeLocation readWithFewerHitsGenomeLocation;
unsigned whichSetPair;
unsigned seedOffset;
unsigned bestPossibleScore;
void init(GenomeLocation readWithFewerHitsGenomeLocation_, unsigned whichSetPair_, unsigned scoringMateCandidateIndex_, unsigned seedOffset_,
unsigned bestPossibleScore_, ScoringCandidate *scoreListNext_)
{
readWithFewerHitsGenomeLocation = readWithFewerHitsGenomeLocation_;
whichSetPair = whichSetPair_;
_ASSERT(whichSetPair < NUM_SET_PAIRS); // You wouldn't think this would be necessary, but...
scoringMateCandidateIndex = scoringMateCandidateIndex_;
seedOffset = seedOffset_;
bestPossibleScore = bestPossibleScore_;
scoreListNext = scoreListNext_;
mergeAnchor = NULL;
}
};
//
// A pool of scoring candidates. For each alignment call, we free them all by resetting lowestFreeScoringCandidatePoolEntry to 0,
// and then fill in the content when they're initialized. This means that for alignments with few candidates we'll be using the same
// entries over and over, so they're likely to be in the cache. We have maxK * maxSeeds * 2 of these in the pool, so we can't possibly run
// out. We rely on their being allocated in descending genome order within a set pair.
//
ScoringCandidate *scoringCandidatePool;
unsigned scoringCandidatePoolSize;
unsigned lowestFreeScoringCandidatePoolEntry;
//
// maxK + 1 lists of Scoring Candidates. The lists correspond to bestPossibleScore for the candidate and its best mate.
//
ScoringCandidate **scoringCandidates;
//
// The scoring mates. The each set scoringCandidatePoolSize / 2.
//
ScoringMateCandidate * scoringMateCandidates[NUM_SET_PAIRS];
unsigned lowestFreeScoringMateCandidate[NUM_SET_PAIRS];
//
// Merge anchors. Again, we allocate an upper bound number of them, which is the same as the number of scoring candidates.
//
MergeAnchor *mergeAnchorPool;
unsigned firstFreeMergeAnchor;
unsigned mergeAnchorPoolSize;
struct HitsPerContigCounts {
_int64 epoch; // Rather than zeroing this whole array every time, we just bump the epoch number; results with an old epoch are considered zero
int hits;
};
HitsPerContigCounts *hitsPerContigCounts; // How many alignments are we reporting for each contig. Used to implement -mpc, otheriwse unallocated.
int maxSecondaryAlignmentsPerContig;
_int64 contigCountEpoch;
};
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