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#include "BitUtil.h"
#include "DataLayer/Options.h"
#include "FMIndex.h"
#include "FastaIndex.h"
#include "FastaInterleave.h"
#include "FastaReader.h"
#include "IOUtil.h"
#include "MemoryUtil.h"
#include "SAM.h"
#include "StringUtil.h"
#include "Uncompress.h"
#include <boost/algorithm/string/join.hpp>
#include <boost/tuple/tuple.hpp>
#include <algorithm>
#include <cassert>
#include <cctype> // for toupper
#include <cstdlib>
#include <getopt.h>
#include <iostream>
#include <stdint.h>
#include <utility>
#include <queue>
#if _OPENMP
# include <omp.h>
#endif
#include "DataBase/Options.h"
#include "DataBase/DB.h"
using namespace std;
using namespace boost;
using namespace boost::algorithm;
#define PROGRAM "abyss-map"
DB db;
static const char VERSION_MESSAGE[] =
PROGRAM " (" PACKAGE_NAME ") " VERSION "\n"
"Written by Shaun Jackman.\n"
"\n"
"Copyright 2014 Canada's Michael Smith Genome Sciences Centre\n";
static const char USAGE_MESSAGE[] =
"Usage: " PROGRAM " [OPTION]... QUERY... TARGET\n"
"Map the sequences of the files QUERY to those of the file TARGET.\n"
"The index files TARGET.fai and TARGET.fm will be used if present.\n"
"\n"
" Options:\n"
"\n"
" -l, --min-align=N find matches at least N bp [1]\n"
" -j, --threads=N use N parallel threads [1]\n"
" -C, --append-comment append the FASTA/FASTQ comment to the SAM tags\n"
" -s, --sample=N sample the suffix array [1]\n"
" -d, --dup identify and print duplicate sequence\n"
" IDs between QUERY and TARGET\n"
" --order print alignments in the same order as\n"
" read from QUERY\n"
" --no-order print alignments ASAP [default]\n"
" --multi Align unaligned segments of primary\n"
" alignment\n"
" --no-multi don't Align unaligned segments [default]\n"
" --SS expect contigs to be oriented correctly\n"
" --no-SS no assumption about contig orientation\n"
" --rc map the sequence and its reverse complement [default]\n"
" --no-rc do not map the reverse complement sequence\n"
" -a, --alphabet=STRING use the alphabet STRING [-ACGT]\n"
" --alpha equivalent to --no-rc -a' ABCDEFGHIJKLMNOPQRSTUVWXYZ'\n"
" --dna equivalent to --rc -a'-ACGT'\n"
" --protein equivalent to --no-rc -a'#*ACDEFGHIKLMNPQRSTVWY'\n"
" --chastity discard unchaste reads\n"
" --no-chastity do not discard unchaste reads [default]\n"
" -v, --verbose display verbose output\n"
" --help display this help and exit\n"
" --version output version information and exit\n"
" --db=FILE specify path of database repository in FILE\n"
" --library=NAME specify library NAME for database\n"
" --strain=NAME specify strain NAME for database\n"
" --species=NAME specify species NAME for database\n"
"\n"
"Report bugs to <" PACKAGE_BUGREPORT ">.\n";
namespace opt {
string db;
dbVars metaVars;
/** Find matches at least k bp. */
static unsigned k;
/** Append the FASTA/FASTQ comment to the SAM tags. */
static int appendComment;
/** Sample the suffix array. */
static unsigned sampleSA;
/** The number of parallel threads. */
static unsigned threads = 1;
/** Run a strand-specific RNA-Seq alignments. */
static int ss;
/** Do not map the sequence's reverse complement. */
static int norc;
/** The alphabet. */
static string alphabet = "-ACGT";
/** Identify duplicate and subsumed sequences. */
static bool dup = false;
/** Align unaligned segments of primary alignment. */
static int multi;
/** Ensure output order matches input order. */
static int order;
/** Verbose output. */
static int verbose;
}
// for sqlite params
static bool haveDbParam(false);
static const char shortopts[] = "Cj:k:l:s:dv";
enum { OPT_HELP = 1, OPT_VERSION,
OPT_ALPHA, OPT_DNA, OPT_PROTEIN,
OPT_DB, OPT_LIBRARY, OPT_STRAIN, OPT_SPECIES,
};
static const struct option longopts[] = {
{ "append-comment", no_argument, NULL, 'C' },
{ "sample", required_argument, NULL, 's' },
{ "min-align", required_argument, NULL, 'l' },
{ "dup", no_argument, NULL, 'd' },
{ "threads", required_argument, NULL, 'j' },
{ "order", no_argument, &opt::order, 1 },
{ "no-order", no_argument, &opt::order, 0 },
{ "multi", no_argument, &opt::multi, 1 },
{ "no-multi", no_argument, &opt::multi, 0 },
{ "SS", no_argument, &opt::ss, 1 },
{ "no-SS", no_argument, &opt::ss, 0 },
{ "rc", no_argument, &opt::norc, 0 },
{ "no-rc", no_argument, &opt::norc, 1 },
{ "alphabet", optional_argument, NULL, 'a' },
{ "alpha", optional_argument, NULL, OPT_ALPHA },
{ "dna", optional_argument, NULL, OPT_DNA },
{ "protein", optional_argument, NULL, OPT_PROTEIN },
{ "decompress", no_argument, NULL, 'd' },
{ "verbose", no_argument, NULL, 'v' },
{ "chastity", no_argument, &opt::chastityFilter, 1 },
{ "no-chastity", no_argument, &opt::chastityFilter, 0 },
{ "help", no_argument, NULL, OPT_HELP },
{ "version", no_argument, NULL, OPT_VERSION },
{ "db", required_argument, NULL, OPT_DB },
{ "library", required_argument, NULL, OPT_LIBRARY },
{ "strain", required_argument, NULL, OPT_STRAIN },
{ "species", required_argument, NULL, OPT_SPECIES },
{ NULL, 0, NULL, 0 }
};
/** Counts. */
static struct {
unsigned unique;
unsigned multimapped;
unsigned unmapped;
unsigned suboptimal;
unsigned subunmapped;
} g_count;
typedef FMIndex::Match Match;
#if SAM_SEQ_QUAL
static string toXA(const FastaIndex& faIndex,
const FMIndex& fmIndex, const Match& m, bool rc,
unsigned qlength, unsigned seq_start)
{
if (m.size() == 0)
return "";
FastaIndex::SeqPos seqPos = faIndex[fmIndex[m.l]];
string rname = seqPos.get<0>().id;
int pos = seqPos.get<1>() + 1;
// Set the mapq to the alignment score.
assert(m.qstart < m.qend);
unsigned matches = m.qend - m.qstart;
unsigned qstart = seq_start + m.qstart;
unsigned qend = m.qend + seq_start;
unsigned short flag = rc ? SAMAlignment::FREVERSE : 0;
ostringstream ss;
if (qstart > 0)
ss << qstart << 'S';
ss << matches << 'M';
if (qend < qlength)
ss << qlength - qend << 'S';
string cigar = ss.str();
stringstream xa_tag;
xa_tag << rname << ',' << pos << ',' << cigar << ",0," << flag;
return xa_tag.str();
}
#endif
/** Return a SAM record of the specified match. */
static SAMRecord toSAM(const FastaIndex& faIndex,
const FMIndex& fmIndex, const Match& m, bool rc,
unsigned qlength)
{
SAMRecord a;
if (m.size() == 0) {
// No hit.
a.rname = "*";
a.pos = -1;
a.flag = SAMAlignment::FUNMAP;
a.mapq = 0;
a.cigar = "*";
} else {
FastaIndex::SeqPos seqPos = faIndex[fmIndex[m.l]];
a.rname = seqPos.get<0>().id;
a.pos = seqPos.get<1>();
a.flag = rc ? SAMAlignment::FREVERSE : 0;
// Set the mapq to the alignment score.
assert(m.qstart < m.qend);
unsigned matches = m.qend - m.qstart;
assert (m.num != 0);
a.mapq = m.size() > 1 || m.num > 1 ? 0 : min(matches, 254U);
ostringstream ss;
if (m.qstart > 0)
ss << m.qstart << 'S';
ss << matches << 'M';
if (m.qend < qlength)
ss << qlength - m.qend << 'S';
a.cigar = ss.str();
}
a.mrnm = "*";
a.mpos = -1;
a.isize = 0;
return a;
}
/** Return the position of the current contig. */
static size_t getMyPos(const Match& m, const FastaIndex& faIndex,
const FMIndex& fmIndex, const string& id)
{
for (size_t i = m.l; i < m.u; i++) {
if (faIndex[fmIndex[i]].get<0>().id == id)
return fmIndex[i];
}
return fmIndex[m.l];
}
/** Return the earlies position of all contigs in m. */
static size_t getMinPos(const Match& m, size_t maxLen,
const FastaIndex& faIndex, const FMIndex& fmIndex)
{
size_t minPos = numeric_limits<size_t>::max();
for (size_t i = m.l; i < m.u; i++) {
size_t pos = fmIndex[i];
if (faIndex[pos].get<0>().size == maxLen && pos < minPos)
minPos = fmIndex[i];
}
return minPos;
}
/** Return the largest length of all contig in m. */
static size_t getMaxLen(const Match& m, const FastaIndex& faIndex,
const FMIndex& fmIndex)
{
size_t maxLen = 0;
for (size_t i = m.l; i < m.u; i++) {
size_t len = faIndex[fmIndex[i]].get<0>().size;
if (len > maxLen)
maxLen = len;
}
return maxLen;
}
/** Print the current contig id if it is not the lartest and earliest
* contig in m. */
static void printDuplicates(const Match& m, const Match& rcm,
const FastaIndex& faIndex, const FMIndex& fmIndex,
const FastqRecord& rec)
{
size_t myLen = m.qspan();
size_t maxLen;
if (opt::ss || opt::norc)
maxLen = getMaxLen(m, faIndex, fmIndex);
else
maxLen = max(getMaxLen(m, faIndex, fmIndex),
getMaxLen(rcm, faIndex, fmIndex));
if (myLen < maxLen) {
#pragma omp atomic
g_count.multimapped++;
#pragma omp critical(cout)
{
cout << rec.id << '\n';
assert_good(cout, "stdout");
}
return;
}
size_t myPos = getMyPos(m, faIndex, fmIndex, rec.id);
size_t minPos;
if (opt::ss || opt::norc)
minPos = getMinPos(m, maxLen, faIndex, fmIndex);
else
minPos = min(getMinPos(m, maxLen, faIndex, fmIndex),
getMinPos(rcm, maxLen, faIndex, fmIndex));
if (myPos > minPos) {
#pragma omp atomic
g_count.multimapped++;
#pragma omp critical(cout)
{
cout << rec.id << '\n';
assert_good(cout, "stdout");
}
}
#pragma omp atomic
g_count.unique++;
return;
}
pair<Match, Match> findMatch(const FMIndex& fmIndex, const string& seq)
{
Match m = fmIndex.find(seq,
opt::dup ? seq.length() : opt::k);
if (opt::norc)
return make_pair(m, Match());
string rcqseq = reverseComplement(seq);
Match rcm;
if (opt::ss)
rcm = fmIndex.find(rcqseq,
opt::dup ? rcqseq.length() : opt::k);
else
rcm = fmIndex.find(rcqseq,
opt::dup ? rcqseq.length() : m.qspan());
return make_pair(m, rcm);
}
static queue<string> g_pq;
/** Return the mapping of the specified sequence. */
static void find(const FastaIndex& faIndex, const FMIndex& fmIndex,
const FastqRecord& rec)
{
if (rec.seq.empty()) {
cerr << PROGRAM ": error: "
"the sequence `" << rec.id << "' is empty\n";
exit(EXIT_FAILURE);
}
Match m, rcm;
tie(m, rcm) = findMatch(fmIndex, rec.seq);
if (opt::dup) {
printDuplicates(m, rcm, faIndex, fmIndex, rec);
return;
}
bool rc;
if (opt::ss) {
rc = rec.id.size() > 2
&& rec.id.substr(rec.id.size()-2) == "/1";
bool prc = rcm.qspan() > m.qspan();
if (prc != rc && ((rc && rcm.size() > 0)
|| (!rc && m.size() > 0)))
#pragma omp atomic
g_count.suboptimal++;
if (prc != rc && ((rc && rcm.size() == 0 && m.size() > 0)
|| (!rc && m.size() == 0 && rcm.size() > 0)))
#pragma omp atomic
g_count.subunmapped++;
} else {
rc = rcm.qspan() > m.qspan();
// if both matches are the same length, sum up the number of times
// each were seen.
if (rcm.qspan() == m.qspan())
rc ? rcm.num += m.num : m.num += rcm.num;
}
vector<string> alts;
Match mm = rc ? rcm : m;
string mseq = rc ? reverseComplement(rec.seq) : rec.seq;
#if SAM_SEQ_QUAL
if (opt::multi) {
if (mm.qstart > 0) {
string seq = mseq.substr(0, mm.qstart);
Match m1, rcm1;
tie(m1, rcm1) = findMatch(fmIndex, seq);
bool rc1 = rcm1.qspan() > m1.qspan();
string xa = toXA(faIndex, fmIndex, rc1 ? rcm1 : m1,
rc ^ rc1, mseq.size(), 0);
if (xa != "")
alts.push_back(xa);
}
if (mm.qend < mseq.size()) {
string seq =
mseq.substr(mm.qend, mseq.length() - mm.qend);
Match m2, rcm2;
tie(m2, rcm2) = findMatch(fmIndex, seq);
bool rc2 = rcm2.qspan() > m2.qspan();
string xa = toXA(faIndex, fmIndex, rc2 ? rcm2 : m2,
rc ^ rc2, mseq.size(), mm.qend);
if (xa != "")
alts.push_back(xa);
}
}
#endif
SAMRecord sam = toSAM(faIndex, fmIndex, mm, rc,
rec.seq.size());
if (rec.id[0] == '@') {
cerr << PROGRAM ": error: "
"the query ID `" << rec.id << "' is invalid since it "
"begins with `@'\n";
exit(EXIT_FAILURE);
}
sam.qname = rec.id;
#if SAM_SEQ_QUAL
sam.seq = mseq;
sam.qual = rec.qual.empty() ? "*" : rec.qual;
if (rc)
reverse(sam.qual.begin(), sam.qual.end());
#endif
bool print = opt::order == 0;
do {
#pragma omp critical(cout)
{
#pragma omp critical(g_pq)
if (!print) {
print = g_pq.front() == rec.id;
if (print)
g_pq.pop();
}
if (print) {
cout << sam;
if (opt::appendComment && !rec.comment.empty()) {
// Output the FASTQ comment, which should be formatted as SAM tags.
cout << '\t' << rec.comment;
} else if (startsWith(rec.comment, "BX:Z:")) {
// Output the BX tag if it's the first tag.
size_t i = rec.comment.find_first_of("\t ");
if (i == string::npos)
i = rec.comment.size();
cout << '\t';
cout.write(rec.comment.data(), i);
}
#if SAM_SEQ_QUAL
if (alts.size() > 0)
cout << "\tXA:Z:" << join(alts, ";");
#endif
cout << '\n';
assert_good(cout, "stdout");
}
}
} while (!print); // spinlock :(
if (sam.isUnmapped())
#pragma omp atomic
g_count.unmapped++;
else if (sam.mapq == 0)
#pragma omp atomic
g_count.multimapped++;
else
#pragma omp atomic
g_count.unique++;
}
/** Map the sequences of the specified file. */
static void find(const FastaIndex& faIndex, const FMIndex& fmIndex,
FastaInterleave& in)
{
#pragma omp parallel
for (FastqRecord rec;;) {
bool good;
#pragma omp critical(in)
{
good = in >> rec;
if (opt::order) {
#pragma omp critical(g_pq)
g_pq.push(rec.id);
}
}
if (good)
find(faIndex, fmIndex, rec);
else
break;
}
assert(in.eof());
}
/** Build an FM index of the specified file. */
static void buildFMIndex(FMIndex& fm, const char* path)
{
if (opt::verbose > 0)
std::cerr << "Reading `" << path << "'...\n";
std::vector<FMIndex::value_type> s;
readFile(path, s);
uint64_t MAX_SIZE = numeric_limits<FMIndex::sais_size_type>::max();
if (s.size() > MAX_SIZE) {
std::cerr << PROGRAM << ": `" << path << "', "
<< toSI(s.size())
<< "B, must be smaller than "
<< toSI(MAX_SIZE) << "B\n";
exit(EXIT_FAILURE);
}
transform(s.begin(), s.end(), s.begin(), ::toupper);
fm.setAlphabet(opt::alphabet);
fm.assign(s.begin(), s.end());
}
/** Return the size of the specified file. */
static streampos fileSize(const string& path)
{
std::ifstream in(path.c_str());
assert_good(in, path);
in.seekg(0, std::ios::end);
assert_good(in, path);
return in.tellg();
}
/** Check that the indexes are up to date. */
static void checkIndexes(const string& path,
const FMIndex& fmIndex, const FastaIndex& faIndex)
{
size_t fastaFileSize = fileSize(path);
if (fmIndex.size() != fastaFileSize) {
cerr << PROGRAM ": `" << path << "': "
"The size of the FM-index, "
<< fmIndex.size()
<< " B, does not match the size of the FASTA file, "
<< fastaFileSize << " B. The index is likely stale.\n";
exit(EXIT_FAILURE);
}
if (faIndex.fileSize() != fastaFileSize) {
cerr << PROGRAM ": `" << path << "': "
"The size of the FASTA index, "
<< faIndex.fileSize()
<< " B, does not match the size of the FASTA file, "
<< fastaFileSize << " B. The index is likely stale.\n";
exit(EXIT_FAILURE);
}
}
int main(int argc, char** argv)
{
string commandLine;
{
ostringstream ss;
char** last = argv + argc - 1;
copy(argv, last, ostream_iterator<const char *>(ss, " "));
ss << *last;
commandLine = ss.str();
}
opt::chastityFilter = false;
opt::trimMasked = false;
if (!opt::db.empty())
opt::metaVars.resize(3);
bool die = false;
for (int c; (c = getopt_long(argc, argv,
shortopts, longopts, NULL)) != -1;) {
istringstream arg(optarg != NULL ? optarg : "");
switch (c) {
case '?': die = true; break;
case 'C': opt::appendComment = 1; break;
case 'j': arg >> opt::threads; break;
case 'k': case 'l':
arg >> opt::k;
break;
case 's': arg >> opt::sampleSA; break;
case 'd': opt::dup = true; break;
case 'a':
opt::alphabet = arg.str();
arg.clear(ios::eofbit);
break;
case OPT_ALPHA:
opt::alphabet = " ABCDEFGHIJKLMNOPQRSTUVWXYZ";
opt::norc = true;
break;
case OPT_DNA:
opt::alphabet = "-ACGT";
opt::norc = false;
break;
case OPT_PROTEIN:
opt::alphabet = "#*ACDEFGHIKLMNPQRSTVWY";
opt::norc = true;
break;
case 'v': opt::verbose++; break;
case OPT_HELP:
cout << USAGE_MESSAGE;
exit(EXIT_SUCCESS);
case OPT_VERSION:
cout << VERSION_MESSAGE;
exit(EXIT_SUCCESS);
case OPT_DB:
arg >> opt::db;
haveDbParam = true;
break;
case OPT_LIBRARY:
arg >> opt::metaVars[0];
haveDbParam = true;
break;
case OPT_STRAIN:
arg >> opt::metaVars[1];
haveDbParam = true;
break;
case OPT_SPECIES:
arg >> opt::metaVars[2]; break;
}
if (optarg != NULL && !arg.eof()) {
cerr << PROGRAM ": invalid option: `-"
<< (char)c << optarg << "'\n";
exit(EXIT_FAILURE);
}
}
#ifndef SAM_SEQ_QUAL
# define SAM_SEQ_QUAL 0
#endif
if (opt::multi && !SAM_SEQ_QUAL) {
cerr << PROGRAM ": multiple alignments not supported with "
"this install. Recompile ABySS with `./configure "
"--enable-samseqqual'.\n";
die = true;
}
if (argc - optind < 2) {
cerr << PROGRAM ": missing arguments\n";
die = true;
}
if (die) {
cerr << "Try `" << PROGRAM
<< " --help' for more information.\n";
exit(EXIT_FAILURE);
}
#if _OPENMP
if (opt::threads > 0)
omp_set_num_threads(opt::threads);
#endif
if (!opt::db.empty()) {
init(db, opt::db,
opt::verbose,
PROGRAM,
opt::getCommand(argc, argv),
opt::metaVars);
addToDb(db, "K", opt::k);
addToDb(db, "SS", opt::ss);
}
const char* targetFile(argv[--argc]);
ostringstream ss;
ss << targetFile << ".fm";
string fmPath(ss.str());
ss.str("");
ss << targetFile << ".fai";
string faiPath(ss.str());
ifstream in;
// Read the FASTA index.
FastaIndex faIndex;
in.open(faiPath.c_str());
if (in) {
if (opt::verbose > 0)
cerr << "Reading `" << faiPath << "'...\n";
in >> faIndex;
assert(in.eof());
in.close();
} else {
if (opt::verbose > 0)
cerr << "Reading `" << targetFile << "'...\n";
faIndex.index(targetFile);
}
if (opt::verbose > 0) {
ssize_t bytes = getMemoryUsage();
if (bytes > 0)
cerr << "Using " << toSI(bytes) << "B of memory and "
<< setprecision(3) << (float)bytes / faIndex.size()
<< " B/sequence.\n";
}
// Read the FM index.
FMIndex fmIndex;
in.open(fmPath.c_str());
if (in) {
if (opt::verbose > 0)
cerr << "Reading `" << fmPath << "'...\n";
assert_good(in, fmPath);
in >> fmIndex;
assert_good(in, fmPath);
in.close();
} else
buildFMIndex(fmIndex, targetFile);
if (opt::sampleSA > 1)
fmIndex.sampleSA(opt::sampleSA);
if (opt::verbose > 0) {
size_t bp = fmIndex.size();
cerr << "Read " << toSI(bp) << "B in "
<< faIndex.size() << " contigs.\n";
ssize_t bytes = getMemoryUsage();
if (bytes > 0)
cerr << "Using " << toSI(bytes) << "B of memory and "
<< setprecision(3) << (float)bytes / bp << " B/bp.\n";
}
if (!opt::db.empty())
addToDb(db, "readContigs", faIndex.size());
// Check that the indexes are up to date.
checkIndexes(targetFile, fmIndex, faIndex);
if (!opt::dup) {
// Write the SAM header.
cout << "@HD\tVN:1.4\n"
"@PG\tID:" PROGRAM "\tPN:" PROGRAM "\tVN:" VERSION "\t"
"CL:" << commandLine << '\n';
faIndex.writeSAMHeader(cout);
cout.flush();
assert_good(cout, "stdout");
} else if (opt::verbose > 0)
cerr << "Identifying duplicates.\n";
FastaInterleave fa(argv + optind, argv + argc,
FastaReader::FOLD_CASE);
find(faIndex, fmIndex, fa);
if (opt::verbose > 0) {
size_t unique = g_count.unique;
size_t mapped = unique + g_count.multimapped;
size_t total = mapped + g_count.unmapped;
cerr << "Mapped " << mapped << " of " << total << " reads ("
<< (float)100 * mapped / total << "%)\n"
<< "Mapped " << unique << " of " << total
<< " reads uniquely (" << (float)100 * unique / total
<< "%)\n";
// TODO: This block shouldn't be in a verbose restricted section.
if (!opt::db.empty()) {
addToDb(db, "read_alignments_initial", total);
addToDb(db, "mapped", mapped);
addToDb(db, "mapped_uniq", unique);
addToDb(db, "reads_map_ss", g_count.suboptimal);
}
if (opt::ss) {
cerr << "Mapped " << g_count.suboptimal
<< " (" << (float)100 * g_count.suboptimal / total << "%)"
<< " reads to the opposite strand of the optimal mapping.\n"
<< "Made " << g_count.subunmapped << " ("
<< (float)100 * g_count.subunmapped / total << "%)"
<< " unmapped suboptimal decisions.\n";
}
}
cout.flush();
assert_good(cout, "stdout");
return 0;
}
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