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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 <cmath>
#include <stdio.h>
#include <iostream>
#include <sstream>
#include <string>
#include <stdexcept>
#include <string.h>
#include <fcntl.h>
#include "sstring.h"
#include "pat.h"
#include "filebuf.h"
#include "formats.h"
#include "util.h"
#include "str_util.h"
#include "tokenize.h"
using namespace std;
/**
* Calculate a per-read random seed based on a combination of
* the read data (incl. sequence, name, quals) and the global
* seed in '_randSeed'.
*/
static uint32_t genRandSeed(
const BTDnaString& qry,
const BTString& qual,
const BTString& name,
uint32_t seed)
{
// Calculate a per-read random seed based on a combination of
// the read data (incl. sequence, name, quals) and the global
// seed
uint32_t rseed = (seed + 101) * 59 * 61 * 67 * 71 * 73 * 79 * 83;
size_t qlen = qry.length();
// Throw all the characters of the read into the random seed
for(size_t i = 0; i < qlen; i++) {
int p = (int)qry[i];
assert_leq(p, 4);
size_t off = ((i & 15) << 1);
rseed ^= ((uint32_t)p << off);
}
// Throw all the quality values for the read into the random
// seed
for(size_t i = 0; i < qlen; i++) {
int p = (int)qual[i];
assert_leq(p, 255);
size_t off = ((i & 3) << 3);
rseed ^= (p << off);
}
// Throw all the characters in the read name into the random
// seed
size_t namelen = name.length();
for(size_t i = 0; i < namelen; i++) {
int p = (int)name[i];
if(p == '/') break;
assert_leq(p, 255);
size_t off = ((i & 3) << 3);
rseed ^= (p << off);
}
return rseed;
}
/**
* Return a new dynamically allocated PatternSource for the given
* format, using the given list of strings as the filenames to read
* from or as the sequences themselves (i.e. if -c was used).
*/
PatternSource* PatternSource::patsrcFromStrings(
const PatternParams& p,
const EList<string>& qs)
{
switch(p.format) {
case FASTA: return new FastaPatternSource(qs, p, p.interleaved);
case FASTA_CONT: return new FastaContinuousPatternSource(qs, p);
case RAW: return new RawPatternSource(qs, p);
case FASTQ: return new FastqPatternSource(qs, p, p.interleaved);
case BAM: return new BAMPatternSource(qs, p);
case TAB_MATE5: return new TabbedPatternSource(qs, p, false);
case TAB_MATE6: return new TabbedPatternSource(qs, p, true);
case CMDLINE: return new VectorPatternSource(qs, p);
case QSEQ: return new QseqPatternSource(qs, p);
#ifdef USE_SRA
case SRA_FASTA:
case SRA_FASTQ: return new SRAPatternSource(qs, p);
#endif
default: {
cerr << "Internal error; bad patsrc format: " << p.format << endl;
throw 1;
}
}
}
/**
* Once name/sequence/qualities have been parsed for an
* unpaired read, set all the other key fields of the Read
* struct.
*/
void PatternSourcePerThread::finalize(Read& ra) {
ra.mate = 1;
ra.rdid = buf_.rdid();
ra.seed = genRandSeed(ra.patFw, ra.qual, ra.name, pp_.seed);
ra.finalize();
if(pp_.fixName) {
ra.fixMateName(1);
}
}
/**
* Once name/sequence/qualities have been parsed for a
* paired-end read, set all the other key fields of the Read
* structs.
*/
void PatternSourcePerThread::finalizePair(Read& ra, Read& rb) {
ra.mate = 1;
rb.mate = 2;
ra.rdid = rb.rdid = buf_.rdid();
ra.seed = genRandSeed(ra.patFw, ra.qual, ra.name, pp_.seed);
rb.seed = genRandSeed(rb.patFw, rb.qual, rb.name, pp_.seed);
ra.finalize();
rb.finalize();
if(pp_.fixName) {
ra.fixMateName(1);
rb.fixMateName(2);
}
}
/**
* Get the next paired or unpaired read from the wrapped
* PatternComposer. Returns a pair of bools; first indicates
* whether we were successful, second indicates whether we're
* done.
*/
pair<bool, bool> PatternSourcePerThread::nextReadPair() {
// Prepare batch
if(buf_.exhausted()) {
pair<bool, int> res = nextBatch(); // more parsing needed!
if(res.first && res.second == 0) {
return make_pair(false, true);
}
last_batch_ = res.first;
last_batch_size_ = res.second;
assert_eq(0, buf_.cur_buf_);
} else {
buf_.next(); // advance cursor; no parsing or locking needed
assert_gt(buf_.cur_buf_, 0);
}
// Now fully parse read/pair *outside* the critical section
assert(!buf_.read_a().readOrigBuf.empty());
assert(buf_.read_a().empty());
if(!parse(buf_.read_a(), buf_.read_b())) {
return make_pair(false, false);
}
// Finalize read/pair
if(!buf_.read_b().patFw.empty()) {
trim(buf_.read_a());
trim(buf_.read_b());
finalizePair(buf_.read_a(), buf_.read_b());
} else {
trim(buf_.read_a());
finalize(buf_.read_a());
}
bool this_is_last = buf_.cur_buf_ == static_cast<unsigned int>(last_batch_size_-1);
return make_pair(true, this_is_last ? last_batch_ : false);
}
/**
* The main member function for dispensing pairs of reads or
* singleton reads. Returns true iff ra and rb contain a new
* pair; returns false if ra contains a new unpaired read.
*/
pair<bool, int> SoloPatternComposer::nextBatch(PerThreadReadBuf& pt) {
size_t cur = cur_;
while(cur < src_->size()) {
// Patterns from srca_[cur_] are unpaired
pair<bool, int> res;
do {
res = (*src_)[cur]->nextBatch(
pt,
true, // batch A (or pairs)
lock_); // grab lock below
} while(!res.first && res.second == 0);
if(res.second == 0) {
ThreadSafe ts(mutex_m);
if(cur + 1 > cur_) {
cur_++;
}
cur = cur_;
continue; // on to next pair of PatternSources
}
return res;
}
assert_leq(cur, src_->size());
return make_pair(true, 0);
}
/**
* The main member function for dispensing pairs of reads or
* singleton reads. Returns true iff ra and rb contain a new
* pair; returns false if ra contains a new unpaired read.
*/
pair<bool, int> DualPatternComposer::nextBatch(PerThreadReadBuf& pt) {
// 'cur' indexes the current pair of PatternSources
size_t cur = cur_;
while(cur < srca_->size()) {
if((*srcb_)[cur] == NULL) {
// Patterns from srca_ are unpaired
pair<bool, int> res = (*srca_)[cur]->nextBatch(
pt,
true, // batch A (or pairs)
lock_); // grab lock below
if(res.second == 0 && cur < srca_->size() - 1) {
ThreadSafe ts(mutex_m);
if(cur + 1 > cur_) cur_++;
cur = cur_; // Move on to next PatternSource
continue; // on to next pair of PatternSources
}
return make_pair(res.first && cur == srca_->size() - 1, res.second);
} else {
pair<bool, int> resa, resb;
// Lock to ensure that this thread gets parallel reads
// in the two mate files
{
ThreadSafe ts(mutex_m);
resa = (*srca_)[cur]->nextBatch(
pt,
true, // batch A
false); // don't grab lock below
resb = (*srcb_)[cur]->nextBatch(
pt,
false, // batch B
false); // don't grab lock below
assert_eq((*srca_)[cur]->readCount(),
(*srcb_)[cur]->readCount());
}
if(resa.second < resb.second) {
cerr << "Error, fewer reads in file specified with -1 "
<< "than in file specified with -2" << endl;
throw 1;
} else if(resa.second == 0 && resb.second == 0) {
ThreadSafe ts(mutex_m);
if(cur + 1 > cur_) {
cur_++;
}
cur = cur_; // Move on to next PatternSource
continue; // on to next pair of PatternSources
} else if(resb.second < resa.second) {
cerr << "Error, fewer reads in file specified with -2 "
<< "than in file specified with -1" << endl;
throw 1;
}
assert_eq(resa.first, resb.first);
assert_eq(resa.second, resb.second);
return make_pair(resa.first && cur == srca_->size() - 1, resa.second);
}
}
assert_leq(cur, srca_->size());
return make_pair(true, 0);
}
/**
* Given the values for all of the various arguments used to specify
* the read and quality input, create a list of pattern sources to
* dispense them.
*/
PatternComposer* PatternComposer::setupPatternComposer(
const EList<string>& si, // singles, from argv
const EList<string>& m1, // mate1's, from -1 arg
const EList<string>& m2, // mate2's, from -2 arg
const EList<string>& m12, // both mates on each line, from --12 arg
const EList<string>& q, // qualities associated with singles
const EList<string>& q1, // qualities associated with m1
const EList<string>& q2, // qualities associated with m2
#ifdef USE_SRA
const EList<string>& sra_accs, // SRA accessions
#endif
PatternParams& p, // read-in parameters
bool verbose) // be talkative?
{
EList<PatternSource*>* a = new EList<PatternSource*>();
EList<PatternSource*>* b = new EList<PatternSource*>();
// Create list of pattern sources for paired reads appearing
// interleaved in a single file
for(size_t i = 0; i < m12.size(); i++) {
const EList<string>* qs = &m12;
EList<string> tmp;
if(p.fileParallel) {
// Feed query files one to each PatternSource
qs = &tmp;
tmp.push_back(m12[i]);
assert_eq(1, tmp.size());
}
a->push_back(PatternSource::patsrcFromStrings(p, *qs));
b->push_back(NULL);
p.interleaved = false;
if(!p.fileParallel) {
break;
}
}
#ifdef USE_SRA
for(size_t i = 0; i < sra_accs.size(); i++) {
const EList<string>* qs = &sra_accs;
EList<string> tmp;
if(p.fileParallel) {
// Feed query files one to each PatternSource
qs = &tmp;
tmp.push_back(sra_accs[i]);
assert_eq(1, tmp.size());
}
a->push_back(PatternSource::patsrcFromStrings(p, *qs));
b->push_back(NULL);
if(!p.fileParallel) {
break;
}
}
#endif
// Create list of pattern sources for paired reads
for(size_t i = 0; i < m1.size(); i++) {
const EList<string>* qs = &m1;
EList<string> tmpSeq;
EList<string> tmpQual;
if(p.fileParallel) {
// Feed query files one to each PatternSource
qs = &tmpSeq;
tmpSeq.push_back(m1[i]);
assert_eq(1, tmpSeq.size());
}
a->push_back(PatternSource::patsrcFromStrings(p, *qs));
if(!p.fileParallel) {
break;
}
}
// Create list of pattern sources for paired reads
for(size_t i = 0; i < m2.size(); i++) {
const EList<string>* qs = &m2;
EList<string> tmpSeq;
EList<string> tmpQual;
if(p.fileParallel) {
// Feed query files one to each PatternSource
qs = &tmpSeq;
tmpSeq.push_back(m2[i]);
assert_eq(1, tmpSeq.size());
}
b->push_back(PatternSource::patsrcFromStrings(p, *qs));
if(!p.fileParallel) {
break;
}
}
// All mates/mate files must be paired
assert_eq(a->size(), b->size());
// Create list of pattern sources for the unpaired reads
for(size_t i = 0; i < si.size(); i++) {
const EList<string>* qs = &si;
PatternSource* patsrc = NULL;
EList<string> tmpSeq;
EList<string> tmpQual;
if(p.fileParallel) {
// Feed query files one to each PatternSource
qs = &tmpSeq;
tmpSeq.push_back(si[i]);
assert_eq(1, tmpSeq.size());
}
patsrc = PatternSource::patsrcFromStrings(p, *qs);
assert(patsrc != NULL);
a->push_back(patsrc);
b->push_back(NULL);
if(!p.fileParallel) {
break;
}
}
PatternComposer *patsrc = NULL;
patsrc = new DualPatternComposer(a, b, p);
return patsrc;
}
void PatternComposer::free_EList_pmembers( const EList<PatternSource*> &elist) {
for (size_t i = 0; i < elist.size(); i++)
if (elist[i] != NULL)
delete elist[i];
}
/**
* Fill Read with the sequence, quality and name for the next
* read in the list of read files. This function gets called by
* all the search threads, so we must handle synchronization.
*
* Returns pair<bool, int> where bool indicates whether we're
* completely done, and int indicates how many reads were read.
*/
pair<bool, int> CFilePatternSource::nextBatchImpl(
PerThreadReadBuf& pt,
bool batch_a)
{
bool done = false;
unsigned nread = 0;
pt.setReadId(readCnt_);
while(true) { // loop that moves on to next file when needed
do {
pair<bool, int> ret = nextBatchFromFile(pt, batch_a, nread);
done = ret.first;
nread = ret.second;
} while(!done && nread == 0); // not sure why this would happen
if(done && filecur_ < infiles_.size()) { // finished with this file
open();
resetForNextFile(); // reset state to handle a fresh file
filecur_++;
if(nread == 0 || (nread < pt.max_buf_)) {
continue;
}
done = false;
}
break;
}
assert_geq(nread, 0);
readCnt_ += nread;
return make_pair(done, nread);
}
pair<bool, int> CFilePatternSource::nextBatch(
PerThreadReadBuf& pt,
bool batch_a,
bool lock)
{
if(lock) {
// synchronization at this level because both reading and manipulation of
// current file pointer have to be protected
ThreadSafe ts(mutex);
return nextBatchImpl(pt, batch_a);
} else {
return nextBatchImpl(pt, batch_a);
}
}
/**
* Open the next file in the list of input files.
*/
void CFilePatternSource::open() {
if(is_open_) {
is_open_ = false;
if (compressed_) {
gzclose(zfp_);
zfp_ = NULL;
}
else {
fclose(fp_);
fp_ = NULL;
}
}
while(filecur_ < infiles_.size()) {
if(infiles_[filecur_] == "-") {
// always assume that data from stdin is compressed
compressed_ = true;
int fd = dup(fileno(stdin));
zfp_ = gzdopen(fd, "rb");
if (zfp_ == NULL) {
close(fd);
}
}
else {
const char* filename = infiles_[filecur_].c_str();
int fd = ::open(filename, O_RDONLY);
bool is_fifo = false;
#ifndef _WIN32
struct stat st;
if (fstat(fd, &st) != 0) {
perror("stat");
}
is_fifo = S_ISFIFO(st.st_mode) != 0;
#endif
if (pp_.format != BAM && (is_fifo || is_gzipped_file(fd))) {
zfp_ = gzdopen(fd, "r");
compressed_ = true;
} else {
fp_ = fdopen(fd, "rb");
}
if((compressed_ && zfp_ == NULL) || (!compressed_ && fp_ == NULL)) {
if (fd != -1) {
close(fd);
}
if(!errs_[filecur_]) {
cerr << "Warning: Could not open read file \""
<< filename
<< "\" for reading; skipping..." << endl;
errs_[filecur_] = true;
}
filecur_++;
continue;
}
}
is_open_ = true;
if (compressed_) {
#if ZLIB_VERNUM < 0x1235
cerr << "Warning: gzbuffer added in zlib v1.2.3.5. Unable to change "
"buffer size from default of 8192." << endl;
#else
gzbuffer(zfp_, 128*1024);
#endif
}
else {
setvbuf(fp_, buf_, _IOFBF, 64*1024);
}
return;
}
cerr << "Error: No input read files were valid" << endl;
exit(1);
return;
}
/**
* Constructor for vector pattern source, used when the user has
* specified the input strings on the command line using the -c
* option.
*/
VectorPatternSource::VectorPatternSource(
const EList<string>& seqs,
const PatternParams& p) :
PatternSource(p),
cur_(p.skip),
skip_(p.skip),
paired_(false),
tokbuf_(),
bufs_()
{
// Install sequences in buffers, ready for immediate copying in
// nextBatch(). Formatting of the buffer is just like
// TabbedPatternSource.
const size_t seqslen = seqs.size();
for(size_t i = 0; i < seqslen; i++) {
tokbuf_.clear();
tokenize(seqs[i], ":", tokbuf_, 2);
assert_gt(tokbuf_.size(), 0);
assert_leq(tokbuf_.size(), 2);
// Get another buffer ready
bufs_.expand();
bufs_.back().clear();
// Install name
itoa10<TReadId>(static_cast<TReadId>(i), nametmp_);
bufs_.back().install(nametmp_);
bufs_.back().append('\t');
// Install sequence
bufs_.back().append(tokbuf_[0].c_str());
bufs_.back().append('\t');
// Install qualities
if(tokbuf_.size() > 1) {
bufs_.back().append(tokbuf_[1].c_str());
} else {
const size_t len = tokbuf_[0].length();
for(size_t i = 0; i < len; i++) {
bufs_.back().append('I');
}
}
}
}
/**
* Read next batch. However, batch concept is not very applicable for this
* PatternSource where all the info has already been parsed into the fields
* in the contsructor. This essentially modifies the pt as though we read
* in some number of patterns.
*/
pair<bool, int> VectorPatternSource::nextBatchImpl(
PerThreadReadBuf& pt,
bool batch_a)
{
pt.setReadId(cur_);
EList<Read>& readbuf = batch_a ? pt.bufa_ : pt.bufb_;
size_t readi = 0;
for(; readi < pt.max_buf_ && cur_ < bufs_.size(); readi++, cur_++) {
readbuf[readi].readOrigBuf = bufs_[cur_];
}
readCnt_ += readi;
return make_pair(cur_ == bufs_.size(), readi);
}
pair<bool, int> VectorPatternSource::nextBatch(
PerThreadReadBuf& pt,
bool batch_a,
bool lock)
{
if(lock) {
ThreadSafe ts(mutex);
return nextBatchImpl(pt, batch_a);
} else {
return nextBatchImpl(pt, batch_a);
}
}
/**
* Finishes parsing outside the critical section.
*/
bool VectorPatternSource::parse(Read& ra, Read& rb, TReadId rdid) const {
// Very similar to TabbedPatternSource
// Light parser (nextBatchFromFile) puts unparsed data
// into Read& r, even when the read is paired.
assert(ra.empty());
assert(!ra.readOrigBuf.empty()); // raw data for read/pair is here
int c = '\t';
size_t cur = 0;
const size_t buflen = ra.readOrigBuf.length();
// Loop over the two ends
for(int endi = 0; endi < 2 && c == '\t'; endi++) {
Read& r = ((endi == 0) ? ra : rb);
assert(r.name.empty());
// Parse name if (a) this is the first end, or
// (b) this is tab6
if(endi < 1 || paired_) {
// Parse read name
c = ra.readOrigBuf[cur++];
while(c != '\t' && cur < buflen) {
r.name.append(c);
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
} else if(endi > 0) {
// if this is the second end and we're parsing
// tab5, copy name from first end
rb.name = ra.name;
}
// Parse sequence
assert(r.patFw.empty());
c = ra.readOrigBuf[cur++];
int nchar = 0;
while(c != '\t' && cur < buflen) {
if(isalpha(c)) {
assert_in(toupper(c), "ACGTN");
if(nchar++ >= pp_.trim5) {
assert_neq(0, asc2dnacat[c]);
r.patFw.append(asc2dna[c]); // ascii to int
}
}
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
// record amt trimmed from 5' end due to --trim5
r.trimmed5 = (int)(nchar - r.patFw.length());
// record amt trimmed from 3' end due to --trim3
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
// Parse qualities
assert(r.qual.empty());
c = ra.readOrigBuf[cur++];
int nqual = 0;
while(c != '\t' && c != '\n' && c != '\r') {
if(c == ' ') {
wrongQualityFormat(r.name);
return false;
}
char cadd = charToPhred33(c, false, false);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
if(cur >= buflen) break;
c = ra.readOrigBuf[cur++];
}
if(nchar > nqual) {
tooFewQualities(r.name);
return false;
} else if(nqual > nchar) {
tooManyQualities(r.name);
return false;
}
r.qual.trimEnd(pp_.trim3);
assert(c == '\t' || c == '\n' || c == '\r' || cur >= buflen);
assert_eq(r.patFw.length(), r.qual.length());
}
ra.parsed = true;
if(!rb.parsed && !rb.readOrigBuf.empty()) {
return parse(rb, ra, rdid);
}
return true;
}
/**
* Light-parse a FASTA batch into the given buffer.
*/
pair<bool, int> FastaPatternSource::nextBatchFromFile(
PerThreadReadBuf& pt,
bool batch_a, unsigned readi)
{
int c;
EList<Read>* readbuf = batch_a ? &pt.bufa_ : &pt.bufb_;
if(first_) {
c = getc_wrapper();
if (c == EOF) {
return make_pair(true, 0);
}
while(c == '\r' || c == '\n') {
c = getc_wrapper();
}
if(c != '>') {
cerr << "Error: reads file does not look like a FASTA file" << endl;
throw 1;
}
first_ = false;
}
bool done = false;
// Read until we run out of input or until we've filled the buffer
while (readi < pt.max_buf_ && !done) {
Read::TBuf& buf = (*readbuf)[readi].readOrigBuf;
buf.clear();
buf.append('>');
while(true) {
c = getc_wrapper();
if(c < 0 || c == '>') {
done = c < 0;
break;
}
buf.append(c);
}
if (interleaved_) {
// alternate between read buffers
batch_a = !batch_a;
readbuf = batch_a ? &pt.bufa_ : &pt.bufb_;
// increment read counter after each pair gets read
readi = batch_a ? readi+1 : readi;
} else {
readi++;
}
}
// Immediate EOF case
if(done && (*readbuf)[readi-1].readOrigBuf.length() == 1) {
readi--;
}
return make_pair(done, readi);
}
/**
* Finalize FASTA parsing outside critical section.
*/
bool FastaPatternSource::parse(Read& r, Read& rb, TReadId rdid) const {
// We assume the light parser has put the raw data for the separate ends
// into separate Read objects. That doesn't have to be the case, but
// that's how we've chosen to do it for FastqPatternSource
assert(!r.readOrigBuf.empty());
assert(r.empty());
int c = -1;
size_t cur = 1;
const size_t buflen = r.readOrigBuf.length();
// Parse read name
assert(r.name.empty());
while(cur < buflen) {
c = r.readOrigBuf[cur++];
if(c == '\n' || c == '\r') {
do {
c = r.readOrigBuf[cur++];
} while((c == '\n' || c == '\r') && cur < buflen);
break;
}
r.name.append(c);
}
if(cur >= buflen) {
return false; // FASTA ended prematurely
}
// Parse sequence
int nchar = 0;
assert(r.patFw.empty());
assert(c != '\n' && c != '\r');
assert_lt(cur, buflen);
while(cur < buflen) {
if(c == '.') {
c = 'N';
}
if(isalpha(c)) {
// If it's past the 5'-end trim point
if(nchar++ >= pp_.trim5) {
r.patFw.append(asc2dna[c]);
}
}
assert_lt(cur, buflen);
c = r.readOrigBuf[cur++];
if ((c == '\n' || c == '\r')
&& cur < buflen
&& r.readOrigBuf[cur] != '>') {
c = r.readOrigBuf[cur++];
}
}
r.trimmed5 = (int)(nchar - r.patFw.length());
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
for(size_t i = 0; i < r.patFw.length(); i++) {
r.qual.append('I');
}
// Set up a default name if one hasn't been set
if(r.name.empty()) {
char cbuf[20];
itoa10<TReadId>(static_cast<TReadId>(rdid), cbuf);
r.name.install(cbuf);
}
r.parsed = true;
if(!rb.parsed && !rb.readOrigBuf.empty()) {
return parse(rb, r, rdid);
}
return true;
}
/**
* Light-parse a FASTA-continuous batch into the given buffer.
* This is trickier for FASTA-continuous than for other formats,
* for several reasons:
*
* 1. Reads are substrings of a longer FASTA input string
* 2. Reads may overlap w/r/t the longer FASTA string
* 3. Read names depend on the most recently observed FASTA
* record name
*/
pair<bool, int> FastaContinuousPatternSource::nextBatchFromFile(
PerThreadReadBuf& pt,
bool batch_a, unsigned readi)
{
int c = -1;
EList<Read>& readbuf = batch_a ? pt.bufa_ : pt.bufb_;
while(readi < pt.max_buf_) {
c = getc_wrapper();
if(c < 0) {
break;
}
if(c == '>') {
resetForNextFile();
c = getc_wrapper();
bool sawSpace = false;
while(c != '\n' && c != '\r') {
if(!sawSpace) {
sawSpace = isspace(c);
}
if(!sawSpace) {
name_prefix_buf_.append(c);
}
c = getc_wrapper();
}
while(c == '\n' || c == '\r') {
c = getc_wrapper();
}
if(c < 0) {
break;
}
name_prefix_buf_.append('_');
}
int cat = asc2dnacat[c];
if(cat >= 2) c = 'N';
if(cat == 0) {
// Non-DNA, non-IUPAC char; skip
continue;
} else {
// DNA char
buf_[bufCur_++] = c;
if(bufCur_ == 1024) {
bufCur_ = 0; // wrap around circular buf
}
if(eat_ > 0) {
eat_--;
// Try to keep cur_ aligned with the offset
// into the reference; that lets us see where
// the sampling gaps are by looking at the read
// name
if(!beginning_) {
cur_++;
}
continue;
}
// install name
readbuf[readi].readOrigBuf = name_prefix_buf_;
itoa10<TReadId>(cur_ - last_, name_int_buf_);
readbuf[readi].readOrigBuf.append(name_int_buf_);
readbuf[readi].readOrigBuf.append('\t');
// install sequence
for(size_t i = 0; i < length_; i++) {
if(length_ - i <= bufCur_) {
c = buf_[bufCur_ - (length_ - i)];
} else {
// Rotate
c = buf_[bufCur_ - (length_ - i) + 1024];
}
readbuf[readi].readOrigBuf.append(c);
}
eat_ = freq_-1;
cur_++;
beginning_ = false;
readi++;
}
}
return make_pair(c < 0, readi);
}
/**
* Finalize FASTA-continuous parsing outside critical section.
*/
bool FastaContinuousPatternSource::parse(
Read& ra,
Read& rb,
TReadId rdid) const
{
// Light parser (nextBatchFromFile) puts unparsed data
// into Read& r, even when the read is paired.
assert(ra.empty());
assert(rb.empty());
assert(!ra.readOrigBuf.empty()); // raw data for read/pair is here
assert(rb.readOrigBuf.empty());
int c = '\t';
size_t cur = 0;
const size_t buflen = ra.readOrigBuf.length();
// Parse read name
c = ra.readOrigBuf[cur++];
while(c != '\t' && cur < buflen) {
ra.name.append(c);
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
// Parse sequence
assert(ra.patFw.empty());
int nchar = 0;
while(cur < buflen) {
c = ra.readOrigBuf[cur++];
if(isalpha(c)) {
assert_in(toupper(c), "ACGTN");
if(nchar++ >= pp_.trim5) {
assert_neq(0, asc2dnacat[c]);
ra.patFw.append(asc2dna[c]); // ascii to int
}
}
}
// record amt trimmed from 5' end due to --trim5
ra.trimmed5 = (int)(nchar - ra.patFw.length());
// record amt trimmed from 3' end due to --trim3
ra.trimmed3 = (int)(ra.patFw.trimEnd(pp_.trim3));
// Make fake qualities
assert(ra.qual.empty());
const size_t len = ra.patFw.length();
for(size_t i = 0; i < len; i++) {
ra.qual.append('I');
}
return true;
}
/**
* "Light" parser. This is inside the critical section, so the key is to do
* just enough parsing so that another function downstream (finalize()) can do
* the rest of the parsing. Really this function's only job is to stick every
* for lines worth of the input file into a buffer (r.readOrigBuf). finalize()
* then parses the contents of r.readOrigBuf later.
*/
pair<bool, int> FastqPatternSource::nextBatchFromFile(
PerThreadReadBuf& pt,
bool batch_a, unsigned readi)
{
int c = -1;
EList<Read>* readbuf = batch_a ? &pt.bufa_ : &pt.bufb_;
if(first_) {
c = getc_wrapper();
if (c == EOF) {
return make_pair(true, 0);
}
while(c == '\r' || c == '\n') {
c = getc_wrapper();
}
if(c != '@') {
cerr << "Error: reads file does not look like a FASTQ file" << endl;
throw 1;
}
first_ = false;
(*readbuf)[readi].readOrigBuf.append('@');
}
bool done = false, aborted = false;
// Read until we run out of input or until we've filled the buffer
while (readi < pt.max_buf_ && !done) {
Read::TBuf& buf = (*readbuf)[readi].readOrigBuf;
int newlines = 4;
while(newlines) {
c = getc_wrapper();
done = c < 0;
if(c == '\n' || (done && newlines == 1)) {
// Saw newline, or EOF that we're
// interpreting as final newline
newlines--;
c = '\n';
} else if(done) {
// account for newline at the end of the file
if (newlines == 4) {
newlines = 0;
}
else {
aborted = true; // Unexpected EOF
}
break;
}
buf.append(c);
}
if (c > 0) {
if (interleaved_) {
// alternate between read buffers
batch_a = !batch_a;
readbuf = batch_a ? &pt.bufa_ : &pt.bufb_;
// increment read counter after each pair gets read
readi = batch_a ? readi+1 : readi;
}
else {
readi++;
}
}
}
if(aborted) {
readi--;
}
return make_pair(done, readi);
}
/**
* Finalize FASTQ parsing outside critical section.
*/
bool FastqPatternSource::parse(Read &r, Read& rb, TReadId rdid) const {
// We assume the light parser has put the raw data for the separate ends
// into separate Read objects. That doesn't have to be the case, but
// that's how we've chosen to do it for FastqPatternSource
assert(!r.readOrigBuf.empty());
assert(r.empty());
int c;
size_t cur = 1;
const size_t buflen = r.readOrigBuf.length();
// Parse read name
assert(r.name.empty());
while(true) {
assert_lt(cur, buflen);
c = r.readOrigBuf[cur++];
if(c == '\n' || c == '\r') {
do {
c = r.readOrigBuf[cur++];
} while(c == '\n' || c == '\r');
break;
}
r.name.append(c);
}
// Parse sequence
int nchar = 0;
assert(r.patFw.empty());
while(c != '+') {
if(c == '.') {
c = 'N';
}
if(isalpha(c)) {
// If it's past the 5'-end trim point
if(nchar++ >= pp_.trim5) {
r.patFw.append(asc2dna[c]);
}
}
assert(cur < r.readOrigBuf.length());
c = r.readOrigBuf[cur++];
}
r.trimmed5 = (int)(nchar - r.patFw.length());
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
assert_eq('+', c);
do {
assert(cur < r.readOrigBuf.length());
c = r.readOrigBuf[cur++];
} while(c != '\n' && c != '\r');
while(cur < buflen && (c == '\n' || c == '\r')) {
c = r.readOrigBuf[cur++];
}
assert(r.qual.empty());
if(nchar > 0) {
int nqual = 0;
if (pp_.intQuals) {
int cur_int = 0;
while(c != '\t' && c != '\n' && c != '\r') {
cur_int *= 10;
cur_int += (int)(c - '0');
c = r.readOrigBuf[cur++];
if(c == ' ' || c == '\t' || c == '\n' || c == '\r') {
char cadd = intToPhred33(cur_int, pp_.solexa64);
cur_int = 0;
assert_geq(cadd, 33);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
}
}
} else {
c = charToPhred33(c, pp_.solexa64, pp_.phred64);
if(nqual++ >= r.trimmed5) {
r.qual.append(c);
}
while(cur < r.readOrigBuf.length()) {
c = r.readOrigBuf[cur++];
if (c == ' ') {
wrongQualityFormat(r.name);
return false;
}
if(c == '\r' || c == '\n') {
break;
}
c = charToPhred33(c, pp_.solexa64, pp_.phred64);
if(nqual++ >= r.trimmed5) {
r.qual.append(c);
}
}
r.qual.trimEnd(r.trimmed3);
if(r.qual.length() < r.patFw.length()) {
tooFewQualities(r.name);
return false;
} else if(r.qual.length() > r.patFw.length()) {
tooManyQualities(r.name);
return false;
}
}
}
// Set up a default name if one hasn't been set
if(r.name.empty()) {
char cbuf[20];
itoa10<TReadId>(static_cast<TReadId>(rdid), cbuf);
r.name.install(cbuf);
}
r.parsed = true;
if(!rb.parsed && !rb.readOrigBuf.empty()) {
return parse(rb, r, rdid);
}
return true;
}
const int BAMPatternSource::offset[] = {
0, //refID
4, //pos
8, //l_read_name
9, //mapq
10, //bin
12, //n_cigar_op
14, //flag
16, //l_seq
20, //next_refID
24, //next_pos
28, //tlen
32, //read_name
};
const uint8_t BAMPatternSource::EOF_MARKER[] = {
0x1f, 0x8b, 0x08, 0x04, 0x00, 0x00, 0x00, 0x00, 0x00, 0xff,
0x06, 0x00, 0x42, 0x43, 0x02, 0x00, 0x1b, 0x00, 0x03, 0x00,
0x00, 0x00, 0x00, 0x00, 0x00, 0x00, 0x00, 0x00
};
uint16_t BAMPatternSource::nextBGZFBlockFromFile(BGZF& b) {
if (fread(&b.hdr, sizeof(b.hdr), 1, fp_) != 1) {
if (feof(fp_))
return 0;
std::cerr << "Error while reading BAM header" << std::endl;
exit(EXIT_FAILURE);
}
uint8_t *extra = new uint8_t[b.hdr.xlen];
if (fread(extra, b.hdr.xlen, 1, fp_) != 1) {
std::cerr << "Error while reading BAM extra subfields" << std::endl;
exit(EXIT_FAILURE);
}
if (memcmp(EOF_MARKER, &b.hdr, sizeof(b.hdr)) == 0 &&
memcmp(EOF_MARKER + sizeof(b.hdr), extra, 6 /* sizeof BAM subfield */) == 0)
{
delete[] extra;
return 0;
}
uint16_t bsize = 0;
for (uint16_t i = 0; i < b.hdr.xlen;) {
if (extra[0] == 66 && extra[1] == 67) {
bsize = *((uint16_t *)(extra + 4));
bsize -= (b.hdr.xlen + 19);
break;
}
i = i + 2;
uint16_t sub_field_len = *((uint16_t *)(extra + 2));
i = i + 2 + sub_field_len;
}
delete[] extra;
if (bsize == 0)
return 0;
if (fread(b.cdata, bsize, 1, fp_) != 1) {
std::cerr << "Error while reading BAM CDATA (compressed data)" << std::endl;
exit(EXIT_FAILURE);
}
if (fread(&b.ftr, sizeof(b.ftr), 1, fp_) != 1) {
std::cerr << "Error while reading BAM footer" << std::endl;
exit(EXIT_FAILURE);
}
return bsize;
}
std::pair<bool, int> BAMPatternSource::nextBatch(PerThreadReadBuf& pt, bool batch_a, bool lock) {
bool done = false;
uint16_t cdata_len;
unsigned nread = 0;
do {
if (bam_batch_indexes_[pt.tid_] >= bam_batches_[pt.tid_].size()) {
BGZF block;
std::vector<uint8_t>& batch = bam_batches_[pt.tid_];
if (lock) {
ThreadSafe ts(mutex);
if (first_) {
// parse the BAM header;
nextBGZFBlockFromFile(block);
first_ = false;
}
cdata_len = nextBGZFBlockFromFile(block);
} else {
if (first_) {
// parse the BAM header
nextBGZFBlockFromFile(block);
first_ = false;
}
cdata_len = nextBGZFBlockFromFile(block);
}
if (cdata_len == 0) {
done = nread == 0;
break;
}
bam_batch_indexes_[pt.tid_] = 0;
batch.resize(block.ftr.isize);
int ret_code = decompress_bgzf_block(&batch[0], block.ftr.isize, block.cdata, cdata_len);
if (ret_code != Z_OK) {
return make_pair(true, 0);
}
uLong crc = crc32(0L, Z_NULL, 0);
crc = crc32(crc, &batch[0], batch.size());
assert(crc == block.ftr.crc32);
}
std::pair<bool, int> ret = get_alignments(pt, batch_a, nread, lock);
done = ret.first;
} while (!done && nread < pt.max_buf_);
if (lock) {
ThreadSafe ts(mutex);
pt.setReadId(readCnt_);
readCnt_ += nread;
} else {
pt.setReadId(readCnt_);
readCnt_ += nread;
}
return make_pair(done, nread);
}
std::pair<bool, int> BAMPatternSource::get_alignments(PerThreadReadBuf& pt, bool batch_a, unsigned& readi, bool lock) {
size_t& i = bam_batch_indexes_[pt.tid_];
bool done = false;
bool read1 = true;
while (readi < pt.max_buf_) {
if (i >= bam_batches_[pt.tid_].size()) {
return make_pair(false, readi);
}
uint16_t flag;
int32_t block_size;
EList<Read>& readbuf = pp_.align_paired_reads && !read1 ? pt.bufb_ : pt.bufa_;
memcpy(&block_size, &bam_batches_[pt.tid_][0] + i, sizeof(block_size));
if (block_size <= 0) {
return make_pair(done, readi);
}
i += sizeof(block_size);
memcpy(&flag, &bam_batches_[pt.tid_][0] + i + offset[BAMField::flag], sizeof(flag));
if (!pp_.align_paired_reads && ((flag & 0x40) != 0 || (flag & 0x80) != 0)) {
readbuf[readi].readOrigBuf.clear();
i += block_size;
continue;
}
if (pp_.align_paired_reads && ((flag & 0x40) == 0 && (flag & 0x80) == 0)) {
readbuf[readi].readOrigBuf.clear();
i += block_size;
continue;
}
if (pp_.align_paired_reads &&
(((flag & 0x40) != 0 && i + block_size == bam_batches_[pt.tid_].size()) ||
((flag & 0x80) != 0 && i == sizeof(block_size))))
{
if (lock) {
ThreadSafe ts(orphan_mates_mutex_);
get_or_store_orhaned_mate(pt.bufa_, pt.bufb_, readi, &bam_batches_[pt.tid_][0] + i, block_size);
i += block_size;
} else {
get_or_store_orhaned_mate(pt.bufa_, pt.bufb_, readi, &bam_batches_[pt.tid_][0] + i, block_size);
i += block_size;
}
} else {
readbuf[readi].readOrigBuf.resize(block_size);
memcpy(readbuf[readi].readOrigBuf.wbuf(), &bam_batches_[pt.tid_][0] + i, block_size);
i += block_size;
read1 = !read1;
readi = (pp_.align_paired_reads &&
pt.bufb_[readi].readOrigBuf.length() == 0) ? readi : readi + 1;
}
}
return make_pair(done, readi);
}
void BAMPatternSource::get_or_store_orhaned_mate(EList<Read>& buf_a, EList<Read>& buf_b, unsigned& readi, const uint8_t *mate, size_t mate_len) {
const char *read_name =
(const char *)(mate + offset[BAMField::read_name]);
size_t i;
uint32_t hash = hash_str(read_name);
orphan_mate_t *empty_slot = NULL;
for (i = 0; i < orphan_mates.size(); i++) {
if (empty_slot == NULL && orphan_mates[i].empty())
empty_slot = &orphan_mates[i];
if (orphan_mates[i].hash == hash)
break;
}
if (i == orphan_mates.size()) {
// vector is full
if (empty_slot == NULL) {
orphan_mates.push_back(orphan_mate_t());
empty_slot = &orphan_mates.back();
}
empty_slot->hash = hash;
if (empty_slot->cap < mate_len) {
delete[] empty_slot->data;
empty_slot->data = NULL;
}
if (empty_slot->data == NULL) {
empty_slot->data = new uint8_t[mate_len];
empty_slot->cap = mate_len;
}
memcpy(empty_slot->data, mate, mate_len);
empty_slot->size = mate_len;
} else {
uint8_t flag;
Read& ra = buf_a[readi];
Read& rb = buf_b[readi];
memcpy(&flag, mate + offset[BAMField::flag], sizeof(flag));
if ((flag & 0x40) != 0) {
ra.readOrigBuf.resize(mate_len);
memcpy(ra.readOrigBuf.wbuf(), mate, mate_len);
rb.readOrigBuf.resize(orphan_mates[i].size);
memcpy(rb.readOrigBuf.wbuf(), orphan_mates[i].data, orphan_mates[i].size);
} else {
rb.readOrigBuf.resize(mate_len);
memcpy(rb.readOrigBuf.wbuf(), mate, mate_len);
ra.readOrigBuf.resize(orphan_mates[i].size);
memcpy(ra.readOrigBuf.wbuf(), orphan_mates[i].data, orphan_mates[i].size);
}
readi++;
orphan_mates[i].reset();
}
}
int BAMPatternSource::decompress_bgzf_block(uint8_t *dst, size_t dst_len, uint8_t *src, size_t src_len) {
z_stream strm;
strm.zalloc = Z_NULL;
strm.zfree = Z_NULL;
strm.opaque = Z_NULL;
strm.avail_in = src_len;
strm.next_in = src;
strm.avail_out = dst_len;
strm.next_out = dst;
int ret = inflateInit2(&strm, -8);
if (ret != Z_OK) {
return ret;
}
ret = inflate(&strm, Z_FINISH);
if (ret != Z_STREAM_END) {
return ret;
}
return inflateEnd(&strm);
}
bool BAMPatternSource::parse(Read& ra, Read& rb, TReadId rdid) const {
uint8_t l_read_name;
int32_t l_seq;
uint16_t n_cigar_op;
const char* buf = ra.readOrigBuf.buf();
int block_size = ra.readOrigBuf.length();
memcpy(&l_read_name, buf + offset[BAMField::l_read_name], sizeof(l_read_name));
memcpy(&n_cigar_op, buf + offset[BAMField::n_cigar_op], sizeof(n_cigar_op));
memcpy(&l_seq, buf + offset[BAMField::l_seq], sizeof(l_seq));
int off = offset[BAMField::read_name];
ra.name.install(buf + off, l_read_name-1);
off += (l_read_name + sizeof(uint32_t) * n_cigar_op);
const char* seq = buf + off;
off += (l_seq+1)/2;
const char* qual = buf + off;
for (int i = 0; i < l_seq; i++) {
if (i < pp_.trim5) {
ra.trimmed5 += 1;
} else {
ra.qual.append(qual[i] + 33);
int base = "=ACMGRSVTWYHKDBN"[static_cast<uint8_t>(seq[i/2]) >> 4*(1-(i%2)) & 0xf];
ra.patFw.append(asc2dna[base]);
}
}
ra.trimmed3 = (int)(ra.patFw.trimEnd(pp_.trim3));
ra.qual.trimEnd(ra.trimmed3);
if (pp_.preserve_tags) {
off += l_seq;
ra.preservedOptFlags.install(buf + off, block_size - off);
}
ra.parsed = true;
if (!rb.parsed && rb.readOrigBuf.length() != 0) {
return parse(rb, ra, rdid);
}
return true;
}
/**
* Light-parse a batch of tabbed-format reads into given buffer.
*/
pair<bool, int> TabbedPatternSource::nextBatchFromFile(
PerThreadReadBuf& pt,
bool batch_a, unsigned readi)
{
int c = getc_wrapper();
while(c >= 0 && (c == '\n' || c == '\r')) {
c = getc_wrapper();
}
EList<Read>& readbuf = batch_a ? pt.bufa_ : pt.bufb_;
// Read until we run out of input or until we've filled the buffer
for(; readi < pt.max_buf_ && c >= 0; readi++) {
readbuf[readi].readOrigBuf.clear();
while(c >= 0 && c != '\n' && c != '\r') {
readbuf[readi].readOrigBuf.append(c);
c = getc_wrapper();
}
while(c >= 0 && (c == '\n' || c == '\r') && readi < pt.max_buf_ - 1) {
c = getc_wrapper();
}
}
return make_pair(c < 0, readi);
}
/**
* Finalize tabbed parsing outside critical section.
*/
bool TabbedPatternSource::parse(Read& ra, Read& rb, TReadId rdid) const {
// Light parser (nextBatchFromFile) puts unparsed data
// into Read& r, even when the read is paired.
assert(ra.empty());
assert(rb.empty());
assert(!ra.readOrigBuf.empty()); // raw data for read/pair is here
assert(rb.readOrigBuf.empty());
int c = '\t';
size_t cur = 0;
const size_t buflen = ra.readOrigBuf.length();
// Loop over the two ends
for(int endi = 0; endi < 2 && c == '\t'; endi++) {
Read& r = ((endi == 0) ? ra : rb);
assert(r.name.empty());
// Parse name if (a) this is the first end, or
// (b) this is tab6
if(endi < 1 || secondName_) {
// Parse read name
c = ra.readOrigBuf[cur++];
while(c != '\t' && cur < buflen) {
r.name.append(c);
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
} else if(endi > 0) {
// if this is the second end and we're parsing
// tab5, copy name from first end
rb.name = ra.name;
}
// Parse sequence
assert(r.patFw.empty());
c = ra.readOrigBuf[cur++];
int nchar = 0;
while(c != '\t' && cur < buflen) {
if(isalpha(c)) {
assert_in(toupper(c), "ACGTN");
if(nchar++ >= pp_.trim5) {
assert_neq(0, asc2dnacat[c]);
r.patFw.append(asc2dna[c]); // ascii to int
}
}
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
// record amt trimmed from 5' end due to --trim5
r.trimmed5 = (int)(nchar - r.patFw.length());
// record amt trimmed from 3' end due to --trim3
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
// Parse qualities
assert(r.qual.empty());
c = ra.readOrigBuf[cur++];
int nqual = 0;
if (pp_.intQuals) {
int cur_int = 0;
while(c != '\t' && c != '\n' && c != '\r' && cur < buflen) {
cur_int *= 10;
cur_int += (int)(c - '0');
c = ra.readOrigBuf[cur++];
if(c == ' ' || c == '\t' || c == '\n' || c == '\r') {
char cadd = intToPhred33(cur_int, pp_.solexa64);
cur_int = 0;
assert_geq(cadd, 33);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
}
}
} else {
while(c != '\t' && c != '\n' && c != '\r') {
if(c == ' ') {
wrongQualityFormat(r.name);
return false;
}
char cadd = charToPhred33(c, pp_.solexa64, pp_.phred64);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
if(cur >= buflen) break;
c = ra.readOrigBuf[cur++];
}
}
if(nchar > nqual) {
tooFewQualities(r.name);
return false;
} else if(nqual > nchar) {
tooManyQualities(r.name);
return false;
}
r.qual.trimEnd(pp_.trim3);
assert(c == '\t' || c == '\n' || c == '\r' || cur >= buflen);
assert_eq(r.patFw.length(), r.qual.length());
}
return true;
}
/**
* Light-parse a batch of raw-format reads into given buffer.
*/
pair<bool, int> RawPatternSource::nextBatchFromFile(
PerThreadReadBuf& pt,
bool batch_a,
unsigned readi)
{
int c = getc_wrapper();
while(c >= 0 && (c == '\n' || c == '\r')) {
c = getc_wrapper();
}
EList<Read>& readbuf = batch_a ? pt.bufa_ : pt.bufb_;
// Read until we run out of input or until we've filled the buffer
for(; readi < pt.max_buf_ && c >= 0; readi++) {
readbuf[readi].readOrigBuf.clear();
while(c >= 0 && c != '\n' && c != '\r') {
readbuf[readi].readOrigBuf.append(c);
c = getc_wrapper();
}
while(c >= 0 && (c == '\n' || c == '\r')) {
c = getc_wrapper();
}
}
// incase a valid character is consumed between batches
if (c >= 0 && c != '\n' && c != '\r') {
ungetc_wrapper(c);
}
return make_pair(c < 0, readi);
}
/**
* Finalize raw parsing outside critical section.
*/
bool RawPatternSource::parse(Read& r, Read& rb, TReadId rdid) const {
assert(r.empty());
assert(!r.readOrigBuf.empty()); // raw data for read/pair is here
int c = '\n';
size_t cur = 0;
const size_t buflen = r.readOrigBuf.length();
// Parse sequence
assert(r.patFw.empty());
int nchar = 0;
while(cur < buflen) {
c = r.readOrigBuf[cur++];
assert(c != '\r' && c != '\n');
if(isalpha(c)) {
assert_in(toupper(c), "ACGTN");
if(nchar++ >= pp_.trim5) {
assert_neq(0, asc2dnacat[c]);
r.patFw.append(asc2dna[c]); // ascii to int
}
}
}
assert_eq(cur, buflen);
// record amt trimmed from 5' end due to --trim5
r.trimmed5 = (int)(nchar - r.patFw.length());
// record amt trimmed from 3' end due to --trim3
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
// Give the name field a dummy value
char cbuf[20];
itoa10<TReadId>(rdid, cbuf);
r.name.install(cbuf);
// Give the base qualities dummy values
assert(r.qual.empty());
const size_t len = r.patFw.length();
for(size_t i = 0; i < len; i++) {
r.qual.append('I');
}
r.parsed = true;
if(!rb.parsed && !rb.readOrigBuf.empty()) {
return parse(rb, r, rdid);
}
return true;
}
void wrongQualityFormat(const BTString& read_name) {
cerr << "Error: Encountered one or more spaces while parsing the quality "
<< "string for read " << read_name << ". If this is a FASTQ file "
<< "with integer (non-ASCII-encoded) qualities, try re-running with "
<< "the --integer-quals option." << endl;
throw 1;
}
void tooFewQualities(const BTString& read_name) {
cerr << "Error: Read " << read_name << " has more read characters than "
<< "quality values." << endl;
throw 1;
}
void tooManyQualities(const BTString& read_name) {
cerr << "Error: Read " << read_name << " has more quality values than read "
<< "characters." << endl;
throw 1;
}
#ifdef USE_SRA
std::pair<bool, int> SRAPatternSource::nextBatchImpl(
PerThreadReadBuf& pt,
bool batch_a)
{
EList<Read>& readbuf = batch_a ? pt.bufa_ : pt.bufb_;
size_t readi = 0;
bool done = false;
for(; readi < pt.max_buf_; readi++) {
if(!sra_its_[pt.tid_]->nextRead() || !sra_its_[pt.tid_]->nextFragment()) {
done = true;
break;
}
const ngs::StringRef rname = sra_its_[pt.tid_]->getReadId();
const ngs::StringRef ra_seq = sra_its_[pt.tid_]->getFragmentBases();
const ngs::StringRef ra_qual = sra_its_[pt.tid_]->getFragmentQualities();
readbuf[readi].readOrigBuf.install(rname.data(), rname.size());
readbuf[readi].readOrigBuf.append('\t');
readbuf[readi].readOrigBuf.append(ra_seq.data(), ra_seq.size());
readbuf[readi].readOrigBuf.append('\t');
readbuf[readi].readOrigBuf.append(ra_qual.data(), ra_qual.size());
if(sra_its_[pt.tid_]->nextFragment()) {
const ngs::StringRef rb_seq = sra_its_[pt.tid_]->getFragmentBases();
const ngs::StringRef rb_qual = sra_its_[pt.tid_]->getFragmentQualities();
readbuf[readi].readOrigBuf.append('\t');
readbuf[readi].readOrigBuf.append(rb_seq.data(), rb_seq.size());
readbuf[readi].readOrigBuf.append('\t');
readbuf[readi].readOrigBuf.append(rb_qual.data(), rb_qual.size());
}
readbuf[readi].readOrigBuf.append('\n');
}
pt.setReadId(readCnt_);
{
ThreadSafe ts(mutex);
readCnt_ += readi;
}
return make_pair(done, readi);
}
/**
* TODO: need to think about whether this can be done in a sensible way
*/
std::pair<bool, int> SRAPatternSource::nextBatch(
PerThreadReadBuf& pt,
bool batch_a,
bool lock)
{
if(lock) {
// synchronization at this level because both reading and manipulation of
// current file pointer have to be protected
return nextBatchImpl(pt, batch_a);
} else {
return nextBatchImpl(pt, batch_a);
}
}
/**
* Finalize tabbed parsing outside critical section.
*/
bool SRAPatternSource::parse(Read& ra, Read& rb, TReadId rdid) const {
// Light parser (nextBatchFromFile) puts unparsed data
// into Read& r, even when the read is paired.
assert(ra.empty());
assert(rb.empty());
assert(!ra.readOrigBuf.empty()); // raw data for read/pair is here
assert(rb.readOrigBuf.empty());
int c = '\t';
size_t cur = 0;
const size_t buflen = ra.readOrigBuf.length();
// Loop over the two ends
for(int endi = 0; endi < 2 && c == '\t'; endi++) {
Read& r = ((endi == 0) ? ra : rb);
assert(r.name.empty());
// Parse name if (a) this is the first end, or
// (b) this is tab6
if(endi < 1) {
// Parse read name
c = ra.readOrigBuf[cur++];
while(c != '\t' && cur < buflen) {
r.name.append(c);
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
} else if(endi > 0) {
// if this is the second end and we're parsing
// tab5, copy name from first end
rb.name = ra.name;
}
// Parse sequence
assert(r.patFw.empty());
c = ra.readOrigBuf[cur++];
int nchar = 0;
while(c != '\t' && cur < buflen) {
if(isalpha(c)) {
assert_in(toupper(c), "ACGTN");
if(nchar++ >= pp_.trim5) {
assert_neq(0, asc2dnacat[c]);
r.patFw.append(asc2dna[c]); // ascii to int
}
}
c = ra.readOrigBuf[cur++];
}
assert_eq('\t', c);
if(cur >= buflen) {
return false; // record ended prematurely
}
// record amt trimmed from 5' end due to --trim5
r.trimmed5 = (int)(nchar - r.patFw.length());
// record amt trimmed from 3' end due to --trim3
r.trimmed3 = (int)(r.patFw.trimEnd(pp_.trim3));
// Parse qualities
assert(r.qual.empty());
c = ra.readOrigBuf[cur++];
int nqual = 0;
if (pp_.intQuals) {
int cur_int = 0;
while(c != '\t' && c != '\n' && c != '\r' && cur < buflen) {
cur_int *= 10;
cur_int += (int)(c - '0');
c = ra.readOrigBuf[cur++];
if(c == ' ' || c == '\t' || c == '\n' || c == '\r') {
char cadd = intToPhred33(cur_int, pp_.solexa64);
cur_int = 0;
assert_geq(cadd, 33);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
}
}
} else {
while(c != '\t' && c != '\n' && c != '\r') {
if(c == ' ') {
wrongQualityFormat(r.name);
return false;
}
char cadd = charToPhred33(c, pp_.solexa64, pp_.phred64);
if(++nqual > pp_.trim5) {
r.qual.append(cadd);
}
if(cur >= buflen) break;
c = ra.readOrigBuf[cur++];
}
}
if(nchar > nqual) {
tooFewQualities(r.name);
return false;
} else if(nqual > nchar) {
tooManyQualities(r.name);
return false;
}
r.qual.trimEnd(pp_.trim3);
assert(c == '\t' || c == '\n' || c == '\r' || cur >= buflen);
assert_eq(r.patFw.length(), r.qual.length());
}
return true;
}
void SRAPatternSource::open() {
const string& sra_acc = sra_accs_[sra_acc_cur_];
string version = "bowtie2";
ncbi::NGS::setAppVersionString(version);
assert(!sra_acc.empty());
try {
// open requested accession using SRA implementation of the API
ngs::ReadCollection sra_run = ncbi::NGS::openReadCollection(sra_acc);
// compute window to iterate through
size_t MAX_ROW = sra_run.getReadCount();
pp_.upto -= pp_.skip;
if (pp_.upto <= MAX_ROW) {
MAX_ROW = pp_.upto;
}
if(MAX_ROW < 0) {
return;
}
size_t window_size = MAX_ROW / sra_its_.size();
size_t remainder = MAX_ROW % sra_its_.size();
size_t i = 0, start = 1;
if (pp_.skip > 0) {
start = pp_.skip + 1;
readCnt_ = pp_.skip;
}
while (i < sra_its_.size()) {
sra_its_[i] = new ngs::ReadIterator(sra_run.getReadRange(start, window_size, ngs::Read::all));
assert(sra_its_[i] != NULL);
i++;
start += window_size;
if (i == sra_its_.size() - 1) {
window_size += remainder;
}
}
} catch(...) {
cerr << "Warning: Could not access \"" << sra_acc << "\" for reading; skipping..." << endl;
}
}
#endif
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