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/*
* Copyright 2011, Ben Langmead <langmea@cs.jhu.edu>
*
* This file is part of Bowtie 2.
*
* Bowtie 2 is free software: you can redistribute it and/or modify
* it under the terms of the GNU General Public License as published by
* the Free Software Foundation, either version 3 of the License, or
* (at your option) any later version.
*
* Bowtie 2 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 General Public License for more details.
*
* You should have received a copy of the GNU General Public License
* along with Bowtie 2. If not, see <http://www.gnu.org/licenses/>.
*/
#include <string>
#include <string.h>
#include "reference.h"
#include "mem_ids.h"
using namespace std;
/**
* Load from .3.gEbwt_ext/.4.gEbwt_ext Bowtie index files.
*/
BitPairReference::BitPairReference(
const string& in,
bool color,
bool sanity,
EList<string>* infiles,
EList<SString<char> >* origs,
bool infilesSeq,
bool useMm,
bool useShmem,
bool mmSweep,
bool verbose,
bool startVerbose) :
buf_(NULL),
sanityBuf_(NULL),
loaded_(true),
sanity_(sanity),
useMm_(useMm),
useShmem_(useShmem),
verbose_(verbose)
{
string s3 = in + ".3." + gEbwt_ext;
string s4 = in + ".4." + gEbwt_ext;
FILE *f3, *f4;
if((f3 = fopen(s3.c_str(), "rb")) == NULL) {
cerr << "Could not open reference-string index file " << s3 << " for reading." << endl;
cerr << "This is most likely because your index was built with an older version" << endl
<< "(<= 0.9.8.1) of bowtie-build. Please re-run bowtie-build to generate a new" << endl
<< "index (or download one from the Bowtie website) and try again." << endl;
loaded_ = false;
return;
}
if((f4 = fopen(s4.c_str(), "rb")) == NULL) {
cerr << "Could not open reference-string index file " << s4 << " for reading." << endl;
loaded_ = false;
return;
}
#ifdef BOWTIE_MM
char *mmFile = NULL;
if(useMm_) {
if(verbose_ || startVerbose) {
cerr << " Memory-mapping reference index file " << s4.c_str() << ": ";
logTime(cerr);
}
struct stat sbuf;
if (stat(s4.c_str(), &sbuf) == -1) {
perror("stat");
cerr << "Error: Could not stat index file " << s4.c_str() << " prior to memory-mapping" << endl;
throw 1;
}
mmFile = (char*)mmap((void *)0, (size_t)sbuf.st_size,
PROT_READ, MAP_SHARED, fileno(f4), 0);
if(mmFile == (void *)(-1) || mmFile == NULL) {
perror("mmap");
cerr << "Error: Could not memory-map the index file " << s4.c_str() << endl;
throw 1;
}
if(mmSweep) {
TIndexOff sum = 0;
for(off_t i = 0; i < sbuf.st_size; i += 1024) {
sum += (TIndexOff) mmFile[i];
}
if(startVerbose) {
cerr << " Swept the memory-mapped ref index file; checksum: " << sum << ": ";
logTime(cerr);
}
}
}
#endif
// Read endianness sentinel, set 'swap'
uint32_t one;
bool swap = false;
one = readIndex<int32_t>(f3, swap);
if(one != 1) {
if(useMm_) {
cerr << "Error: Can't use memory-mapped files when the index is the opposite endianness" << endl;
throw 1;
}
assert_eq(0x1000000, one);
swap = true; // have to endian swap U32s
}
// Read # records
TIndexOffU sz;
sz = readIndex<TIndexOffU>(f3, swap);
if(sz == 0) {
cerr << "Error: number of reference records is 0 in " << s3.c_str() << endl;
throw 1;
}
// Read records
nrefs_ = 0;
// Cumulative count of all unambiguous characters on a per-
// stretch 8-bit alignment (i.e. count of bytes we need to
// allocate in buf_)
TIndexOffU cumsz = 0;
TIndexOffU cumlen = 0;
// For each unambiguous stretch...
for(TIndexOffU i = 0; i < sz; i++) {
recs_.push_back(RefRecord(f3, swap));
if(recs_.back().first) {
// This is the first record for this reference sequence (and the
// last record for the one before)
refRecOffs_.push_back((TIndexOffU)recs_.size()-1);
// refOffs_ links each reference sequence with the total number of
// unambiguous characters preceding it in the pasted reference
refOffs_.push_back(cumsz);
if(nrefs_ > 0) {
// refLens_ links each reference sequence with the total number
// of ambiguous and unambiguous characters in it.
refLens_.push_back(cumlen);
}
cumlen = 0;
nrefs_++;
} else if(i == 0) {
cerr << "First record in reference index file was not marked as "
<< "'first'" << endl;
throw 1;
}
cumUnambig_.push_back(cumsz);
cumRefOff_.push_back(cumlen);
cumsz += recs_.back().len;
cumlen += recs_.back().off;
cumlen += recs_.back().len;
}
if(verbose_ || startVerbose) {
cerr << "Read " << nrefs_ << " reference strings from "
<< sz << " records: ";
logTime(cerr);
}
// Store a cap entry for the end of the last reference seq
refRecOffs_.push_back((TIndexOffU)recs_.size());
refOffs_.push_back(cumsz);
refLens_.push_back(cumlen);
cumUnambig_.push_back(cumsz);
cumRefOff_.push_back(cumlen);
bufSz_ = cumsz;
assert_eq(nrefs_, refLens_.size());
assert_eq(sz, recs_.size());
if (f3 != NULL) fclose(f3); // done with .3.gEbwt_ext file
// Round cumsz up to nearest byte boundary
if((cumsz & 3) != 0) {
cumsz += (4 - (cumsz & 3));
}
bufAllocSz_ = cumsz >> 2;
assert_eq(0, cumsz & 3); // should be rounded up to nearest 4
if(useMm_) {
#ifdef BOWTIE_MM
buf_ = (uint8_t*)mmFile;
if(sanity_) {
FILE *ftmp = fopen(s4.c_str(), "rb");
sanityBuf_ = new uint8_t[cumsz >> 2];
size_t ret = fread(sanityBuf_, 1, cumsz >> 2, ftmp);
if(ret != (cumsz >> 2)) {
cerr << "Only read " << ret << " bytes (out of " << (cumsz >> 2) << ") from reference index file " << s4.c_str() << endl;
throw 1;
}
fclose(ftmp);
for(size_t i = 0; i < (cumsz >> 2); i++) {
assert_eq(sanityBuf_[i], buf_[i]);
}
}
#else
cerr << "Shouldn't be at " << __FILE__ << ":" << __LINE__ << " without BOWTIE_MM defined" << endl;
throw 1;
#endif
} else {
bool shmemLeader = true;
if(!useShmem_) {
// Allocate a buffer to hold the reference string
try {
buf_ = new uint8_t[cumsz >> 2];
if(buf_ == NULL) throw std::bad_alloc();
} catch(std::bad_alloc& e) {
cerr << "Error: Ran out of memory allocating space for the bitpacked reference. Please" << endl
<< "re-run on a computer with more memory." << endl;
throw 1;
}
} else {
shmemLeader = ALLOC_SHARED_U8(
(s4 + "[ref]"), (cumsz >> 2), &buf_,
"ref", (verbose_ || startVerbose));
}
if(shmemLeader) {
// Open the bitpair-encoded reference file
FILE *f4 = fopen(s4.c_str(), "rb");
if(f4 == NULL) {
cerr << "Could not open reference-string index file " << s4.c_str() << " for reading." << endl;
cerr << "This is most likely because your index was built with an older version" << endl
<< "(<= 0.9.8.1) of bowtie-build. Please re-run bowtie-build to generate a new" << endl
<< "index (or download one from the Bowtie website) and try again." << endl;
loaded_ = false;
return;
}
// Read the whole thing in
size_t ret = fread(buf_, 1, cumsz >> 2, f4);
// Didn't read all of it?
if(ret != (cumsz >> 2)) {
cerr << "Only read " << ret << " bytes (out of " << (cumsz >> 2) << ") from reference index file " << s4.c_str() << endl;
throw 1;
}
// Make sure there's no more
char c;
ret = fread(&c, 1, 1, f4);
assert_eq(0, ret); // should have failed
fclose(f4);
#ifdef BOWTIE_SHARED_MEM
if(useShmem_) NOTIFY_SHARED(buf_, (cumsz >> 2));
#endif
} else {
#ifdef BOWTIE_SHARED_MEM
if(useShmem_) WAIT_SHARED(buf_, (cumsz >> 2));
#endif
}
}
// Populate byteToU32_
bool big = currentlyBigEndian();
for(int i = 0; i < 256; i++) {
uint32_t word = 0;
if(big) {
word |= ((i >> 0) & 3) << 24;
word |= ((i >> 2) & 3) << 16;
word |= ((i >> 4) & 3) << 8;
word |= ((i >> 6) & 3) << 0;
} else {
word |= ((i >> 0) & 3) << 0;
word |= ((i >> 2) & 3) << 8;
word |= ((i >> 4) & 3) << 16;
word |= ((i >> 6) & 3) << 24;
}
byteToU32_[i] = word;
}
#ifndef NDEBUG
if(sanity_) {
// Compare the sequence we just read from the compact index
// file to the true reference sequence.
EList<SString<char> > *os; // for holding references
EList<SString<char> > osv(DEBUG_CAT); // for holding ref seqs
EList<SString<char> > osn(DEBUG_CAT); // for holding ref names
EList<size_t> osvLen(DEBUG_CAT); // for holding ref seq lens
EList<size_t> osnLen(DEBUG_CAT); // for holding ref name lens
SStringExpandable<uint32_t> tmp_destU32_;
if(infiles != NULL) {
if(infilesSeq) {
for(size_t i = 0; i < infiles->size(); i++) {
// Remove initial backslash; that's almost
// certainly being used to protect the first
// character of the sequence from getopts (e.g.,
// when the first char is -)
if((*infiles)[i].at(0) == '\\') {
(*infiles)[i].erase(0, 1);
}
osv.push_back(SString<char>((*infiles)[i]));
}
} else {
parseFastas(*infiles, osn, osnLen, osv, osvLen);
}
os = &osv;
} else {
assert(origs != NULL);
os = origs;
}
// Go through the loaded reference files base-by-base and
// sanity check against what we get by calling getBase and
// getStretch
for(size_t i = 0; i < os->size(); i++) {
size_t olen = ((*os)[i]).length();
size_t olenU32 = (olen + 12) / 4;
uint32_t *buf = new uint32_t[olenU32];
uint8_t *bufadj = (uint8_t*)buf;
bufadj += getStretch(buf, i, 0, olen, tmp_destU32_);
for(size_t j = 0; j < olen; j++) {
assert_eq((int)(*os)[i][j], (int)bufadj[j]);
assert_eq((int)(*os)[i][j], (int)getBase(i, j));
}
delete[] buf;
}
}
#endif
}
BitPairReference::~BitPairReference() {
if(buf_ != NULL && !useMm_ && !useShmem_) delete[] buf_;
if(sanityBuf_ != NULL) delete[] sanityBuf_;
}
/**
* Return a single base of the reference. Calling this repeatedly
* is not an efficient way to retrieve bases from the reference;
* use loadStretch() instead.
*
* This implementation scans linearly through the records for the
* unambiguous stretches of the target reference sequence. When
* there are many records, binary search would be more appropriate.
*/
int BitPairReference::getBase(size_t tidx, size_t toff) const {
uint64_t reci = refRecOffs_[tidx]; // first record for target reference sequence
uint64_t recf = refRecOffs_[tidx+1]; // last record (exclusive) for target seq
assert_gt(recf, reci);
uint64_t bufOff = refOffs_[tidx];
uint64_t off = 0;
// For all records pertaining to the target reference sequence...
for(uint64_t i = reci; i < recf; i++) {
assert_geq(toff, off);
off += recs_[i].off;
if(toff < off) {
return 4;
}
assert_geq(toff, off);
uint64_t recOff = off + recs_[i].len;
if(toff < recOff) {
toff -= off;
bufOff += (uint64_t)toff;
assert_lt(bufOff, bufSz_);
const uint64_t bufElt = (bufOff) >> 2;
const uint64_t shift = (bufOff & 3) << 1;
return ((buf_[bufElt] >> shift) & 3);
}
bufOff += recs_[i].len;
off = recOff;
assert_geq(toff, off);
} // end for loop over records
return 4;
}
/**
* Load a stretch of the reference string into memory at 'dest'.
*
* This implementation scans linearly through the records for the
* unambiguous stretches of the target reference sequence. When
* there are many records, binary search would be more appropriate.
*/
int BitPairReference::getStretchNaive(
uint32_t *destU32,
size_t tidx,
size_t toff,
size_t count) const
{
uint8_t *dest = (uint8_t*)destU32;
uint64_t reci = refRecOffs_[tidx]; // first record for target reference sequence
uint64_t recf = refRecOffs_[tidx+1]; // last record (exclusive) for target seq
assert_gt(recf, reci);
uint64_t cur = 0;
uint64_t bufOff = refOffs_[tidx];
uint64_t off = 0;
// For all records pertaining to the target reference sequence...
for(uint64_t i = reci; i < recf; i++) {
assert_geq(toff, off);
off += recs_[i].off;
for(; toff < off && count > 0; toff++) {
dest[cur++] = 4;
count--;
}
if(count == 0) break;
assert_geq(toff, off);
if(toff < off + recs_[i].len) {
bufOff += (TIndexOffU)(toff - off); // move bufOff pointer forward
} else {
bufOff += recs_[i].len;
}
off += recs_[i].len;
for(; toff < off && count > 0; toff++) {
assert_lt(bufOff, bufSz_);
const uint64_t bufElt = (bufOff) >> 2;
const uint64_t shift = (bufOff & 3) << 1;
dest[cur++] = (buf_[bufElt] >> shift) & 3;
bufOff++;
count--;
}
if(count == 0) break;
assert_geq(toff, off);
} // end for loop over records
// In any chars are left after scanning all the records,
// they must be ambiguous
while(count > 0) {
count--;
dest[cur++] = 4;
}
assert_eq(0, count);
return 0;
}
/**
* Load a stretch of the reference string into memory at 'dest'.
*/
int BitPairReference::getStretch(
uint32_t *destU32,
size_t tidx,
size_t toff,
size_t count
ASSERT_ONLY(, SStringExpandable<uint32_t>& destU32_2)) const
{
ASSERT_ONLY(size_t origCount = count);
ASSERT_ONLY(size_t origToff = toff);
if(count == 0) return 0;
uint8_t *dest = (uint8_t*)destU32;
#ifndef NDEBUG
destU32_2.clear();
uint8_t *dest_2 = NULL;
int off2;
if((rand() % 10) == 0) {
destU32_2.resize((origCount >> 2) + 2);
off2 = getStretchNaive(destU32_2.wbuf(), tidx, origToff, origCount);
dest_2 = ((uint8_t*)destU32_2.wbuf()) + off2;
}
#endif
destU32[0] = 0x04040404; // Add Ns, which we might end up using later
uint64_t reci = refRecOffs_[tidx]; // first record for target reference sequence
uint64_t recf = refRecOffs_[tidx+1]; // last record (exclusive) for target seq
assert_gt(recf, reci);
uint64_t cur = 4; // keep a cushion of 4 bases at the beginning
uint64_t bufOff = refOffs_[tidx];
uint64_t off = 0;
int64_t offset = 4;
bool firstStretch = true;
bool binarySearched = false;
uint64_t left = reci;
uint64_t right = recf;
uint64_t mid = 0;
// For all records pertaining to the target reference sequence...
for(uint64_t i = reci; i < recf; i++) {
uint64_t origBufOff = bufOff;
assert_geq(toff, off);
if (firstStretch && recf > reci + 16){
// binary search finds smallest i s.t. toff >= cumRefOff_[i]
while (left < right-1) {
mid = left + ((right - left) >> 1);
if (cumRefOff_[mid] <= toff)
left = mid;
else
right = mid;
}
off = cumRefOff_[left];
bufOff = cumUnambig_[left];
origBufOff = bufOff;
i = left;
assert(cumRefOff_[i+1] == 0 || cumRefOff_[i+1] > toff);
binarySearched = true;
}
off += recs_[i].off; // skip Ns at beginning of stretch
assert_gt(count, 0);
if(toff < off) {
size_t cpycnt = min((size_t)(off - toff), count);
memset(&dest[cur], 4, cpycnt);
count -= cpycnt;
toff += cpycnt;
cur += cpycnt;
if(count == 0) break;
}
assert_geq(toff, off);
if(toff < off + recs_[i].len) {
bufOff += toff - off; // move bufOff pointer forward
} else {
bufOff += recs_[i].len;
}
off += recs_[i].len;
assert(off == cumRefOff_[i+1] || cumRefOff_[i+1] == 0);
assert(!binarySearched || toff < off);
_unused(binarySearched); //make production build happy
if(toff < off) {
if(firstStretch) {
if(toff + 8 < off && count > 8) {
// We already added some Ns, so we have to do
// a fixup at the beginning of the buffer so
// that we can start clobbering at cur >> 2
if(cur & 3) {
offset -= (cur & 3);
}
uint64_t curU32 = cur >> 2;
// Do the initial few bases
if(bufOff & 3) {
const uint64_t bufElt = (bufOff) >> 2;
const int64_t low2 = bufOff & 3;
// Lots of cache misses on the following line
destU32[curU32] = byteToU32_[buf_[bufElt]];
for(int j = 0; j < low2; j++) {
((char *)(&destU32[curU32]))[j] = 4;
}
curU32++;
offset += low2;
const int64_t chars = 4 - low2;
count -= chars;
bufOff += chars;
toff += chars;
}
assert_eq(0, bufOff & 3);
uint64_t bufOffU32 = bufOff >> 2;
uint64_t countLim = count >> 2;
uint64_t offLim = ((off - (toff + 4)) >> 2);
uint64_t lim = min(countLim, offLim);
// Do the fast thing for as far as possible
for(uint64_t j = 0; j < lim; j++) {
// Lots of cache misses on the following line
destU32[curU32] = byteToU32_[buf_[bufOffU32++]];
#ifndef NDEBUG
if(dest_2 != NULL) {
assert_eq(dest[(curU32 << 2) + 0], dest_2[(curU32 << 2) - offset + 0]);
assert_eq(dest[(curU32 << 2) + 1], dest_2[(curU32 << 2) - offset + 1]);
assert_eq(dest[(curU32 << 2) + 2], dest_2[(curU32 << 2) - offset + 2]);
assert_eq(dest[(curU32 << 2) + 3], dest_2[(curU32 << 2) - offset + 3]);
}
#endif
curU32++;
}
toff += (lim << 2);
assert_leq(toff, off);
assert_leq((lim << 2), count);
count -= (lim << 2);
bufOff = bufOffU32 << 2;
cur = curU32 << 2;
}
// Do the slow thing for the rest
for(; toff < off && count > 0; toff++) {
assert_lt(bufOff, bufSz_);
const uint64_t bufElt = (bufOff) >> 2;
const uint64_t shift = (bufOff & 3) << 1;
dest[cur++] = (buf_[bufElt] >> shift) & 3;
bufOff++;
count--;
}
firstStretch = false;
} else {
// Do the slow thing
for(; toff < off && count > 0; toff++) {
assert_lt(bufOff, bufSz_);
const uint64_t bufElt = (bufOff) >> 2;
const uint64_t shift = (bufOff & 3) << 1;
dest[cur++] = (buf_[bufElt] >> shift) & 3;
bufOff++;
count--;
}
}
}
if(count == 0) break;
assert_eq(recs_[i].len, bufOff - origBufOff);
_unused(origBufOff); // make production build happy
assert_geq(toff, off);
} // end for loop over records
// In any chars are left after scanning all the records,
// they must be ambiguous
while(count > 0) {
count--;
dest[cur++] = 4;
}
assert_eq(0, count);
return (int)offset;
}
/**
* Parse the input fasta files, populating the szs list and writing the
* .3.gEbwt_ext and .4.gEbwt_ext portions of the index as we go.
*/
pair<size_t, size_t>
BitPairReference::szsFromFasta(
EList<FileBuf*>& is,
const string& outfile,
bool bigEndian,
const RefReadInParams& refparams,
EList<RefRecord>& szs,
bool sanity)
{
RefReadInParams parms = refparams;
std::pair<size_t, size_t> sztot;
if(!outfile.empty()) {
string file3 = outfile + ".3." + gEbwt_ext;
string file4 = outfile + ".4." + gEbwt_ext;
// Open output stream for the '.3.gEbwt_ext' file which will
// hold the size records.
ofstream fout3(file3.c_str(), ios::binary);
if(!fout3.good()) {
cerr << "Could not open index file for writing: \"" << file3.c_str() << "\"" << endl
<< "Please make sure the directory exists and that permissions allow writing by" << endl
<< "Bowtie." << endl;
throw 1;
}
BitpairOutFileBuf bpout(file4.c_str());
// Read in the sizes of all the unambiguous stretches of the genome
// into a vector of RefRecords. The input streams are reset once
// it's done.
writeIndex<int32_t>(fout3, 1, bigEndian); // endianness sentinel
bool color = parms.color;
if(color) {
parms.color = false;
// Make sure the .3.gEbwt_ext and .4.gEbwt_ext files contain
// nucleotides; not colors
TIndexOff numSeqs = 0;
ASSERT_ONLY(std::pair<size_t, size_t> sztot2 =)
fastaRefReadSizes(is, szs, parms, &bpout, numSeqs);
parms.color = true;
writeIndex<TIndexOffU>(fout3, (TIndexOffU)szs.size(), bigEndian); // write # records
for(size_t i = 0; i < szs.size(); i++) {
szs[i].write(fout3, bigEndian);
}
szs.clear();
// Now read in the colorspace size records; these are
// the ones that were indexed
TIndexOff numSeqs2 = 0;
sztot = fastaRefReadSizes(is, szs, parms, NULL, numSeqs2);
assert_eq(numSeqs, numSeqs2);
assert_eq(sztot2.second, sztot.second + numSeqs);
} else {
TIndexOff numSeqs = 0;
sztot = fastaRefReadSizes(is, szs, parms, &bpout, numSeqs);
writeIndex<TIndexOffU>(fout3, (TIndexOffU)szs.size(), bigEndian); // write # records
for(size_t i = 0; i < szs.size(); i++) szs[i].write(fout3, bigEndian);
}
if(sztot.first == 0) {
cerr << "Error: No unambiguous stretches of characters in the input. Aborting..." << endl;
throw 1;
}
assert_gt(sztot.first, 0);
assert_gt(sztot.second, 0);
bpout.close();
fout3.close();
} else {
// Read in the sizes of all the unambiguous stretches of the
// genome into a vector of RefRecords
TIndexOff numSeqs = 0;
sztot = fastaRefReadSizes(is, szs, parms, NULL, numSeqs);
#ifndef NDEBUG
if(parms.color) {
parms.color = false;
EList<RefRecord> szs2(EBWTB_CAT);
TIndexOff numSeqs2 = 0;
ASSERT_ONLY(std::pair<size_t, size_t> sztot2 =)
fastaRefReadSizes(is, szs2, parms, NULL, numSeqs2);
assert_eq(numSeqs, numSeqs2);
// One less color than base
assert_geq(sztot2.second, sztot.second + numSeqs);
parms.color = true;
}
#endif
}
return sztot;
}
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