1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407
|
/*
* sam.cpp
*
* Created on: Sep 23, 2009
* Author: Ben Langmead
*/
#include <vector>
#include <string>
#include <iostream>
#include "pat.h"
#include "hit.h"
#include "sam.h"
using namespace std;
/**
* Write the SAM header lines.
*/
void SAMHitSink::appendHeaders(OutFileBuf& os,
size_t numRefs,
const vector<string>& refnames,
bool color,
bool nosq,
ReferenceMap *rmap,
const TIndexOffU* plen,
bool fullRef,
bool noQnameTrunc,
const char *cmdline,
const char *rgline)
{
ostringstream ss;
ss << "@HD\tVN:1.0\tSO:unsorted" << endl;
if(!nosq) {
for(size_t i = 0; i < numRefs; i++) {
// RNAME
ss << "@SQ\tSN:";
if(!refnames.empty() && rmap != NULL) {
printUptoWs(ss, rmap->getName(i), !fullRef);
} else if(i < refnames.size()) {
printUptoWs(ss, refnames[i], !fullRef);
} else {
ss << i;
}
ss << "\tLN:" << (plen[i] + (color ? 1 : 0)) << endl;
}
}
if(rgline != NULL) {
ss << "@RG\t" << rgline << endl;
}
ss << "@PG\tID:Bowtie\tVN:" << BOWTIE_VERSION << "\tCL:\"" << cmdline << "\"" << endl;
os.writeString(ss.str());
}
/**
* Append a SAM output record for an unaligned read.
*/
void SAMHitSink::appendAligned(ostream& ss,
const Hit& h,
int mapq,
int xms, // value for XM:I field
const vector<string>* refnames,
ReferenceMap *rmap,
AnnotationMap *amap,
bool fullRef,
bool noQnameTrunc,
int offBase)
{
// QNAME
if(h.mate > 0) {
// truncate final 2 chars
for(int i = 0; i < (int)seqan::length(h.patName)-2; i++) {
if(!noQnameTrunc && isspace((int)h.patName[i])) break;
ss << h.patName[i];
}
} else {
for(int i = 0; i < (int)seqan::length(h.patName); i++) {
if(!noQnameTrunc && isspace((int)h.patName[i])) break;
ss << h.patName[i];
}
}
ss << '\t';
// FLAG
int flags = 0;
if(h.mate == 1) {
flags |= SAM_FLAG_PAIRED | SAM_FLAG_FIRST_IN_PAIR | SAM_FLAG_MAPPED_PAIRED;
} else if(h.mate == 2) {
flags |= SAM_FLAG_PAIRED | SAM_FLAG_SECOND_IN_PAIR | SAM_FLAG_MAPPED_PAIRED;
}
if(!h.fw) flags |= SAM_FLAG_QUERY_STRAND;
if(h.mate > 0 && !h.mfw) flags |= SAM_FLAG_MATE_STRAND;
ss << flags << "\t";
// RNAME
if(refnames != NULL && rmap != NULL) {
printUptoWs(ss, rmap->getName(h.h.first), !fullRef);
} else if(refnames != NULL && h.h.first < refnames->size()) {
printUptoWs(ss, (*refnames)[h.h.first], !fullRef);
} else {
ss << h.h.first;
}
// POS
ss << '\t' << (h.h.second + 1);
// MAPQ
ss << "\t" << mapq;
// CIGAR
ss << '\t' << h.length() << 'M';
// MRNM
if(h.mate > 0) {
ss << "\t=";
} else {
ss << "\t*";
}
// MPOS
if(h.mate > 0) {
ss << '\t' << (h.mh.second + 1);
} else {
ss << "\t0";
}
// ISIZE
ss << '\t';
if(h.mate > 0) {
assert_eq(h.h.first, h.mh.first);
int64_t inslen = 0;
if(h.h.second > h.mh.second) {
inslen = (int64_t)h.h.second - (int64_t)h.mh.second + (int64_t)h.length();
inslen = -inslen;
} else {
inslen = (int64_t)h.mh.second - (int64_t)h.h.second + (int64_t)h.mlen;
}
ss << inslen;
} else {
ss << '0';
}
// SEQ
ss << '\t' << h.patSeq;
// QUAL
ss << '\t' << h.quals;
//
// Optional fields
//
// Always output stratum
ss << "\tXA:i:" << (int)h.stratum;
// Always output cost
//ss << "\tXC:i:" << (int)h.cost;
// Look for SNP annotations falling within the alignment
// Output MD field
size_t len = length(h.patSeq);
int nm = 0;
int run = 0;
ss << "\tMD:Z:";
const FixedBitset<1024> *mms = &h.mms;
ASSERT_ONLY(const String<Dna5>* pat = &h.patSeq);
const vector<char>* refcs = &h.refcs;
if(h.color && false) {
// Disabled: print MD:Z string w/r/t to colors, not letters
mms = &h.cmms;
ASSERT_ONLY(pat = &h.colSeq);
assert_eq(length(h.colSeq), len+1);
len = length(h.colSeq);
refcs = &h.crefcs;
}
if(h.fw) {
for (int i = 0; i < (int)len; ++ i) {
if(mms->test(i)) {
nm++;
// There's a mismatch at this position
assert_gt((int)refcs->size(), i);
char refChar = toupper((*refcs)[i]);
ASSERT_ONLY(char qryChar = (h.fw ? (*pat)[i] : (*pat)[len-i-1]));
assert_neq(refChar, qryChar);
ss << run << refChar;
run = 0;
} else {
run++;
}
}
} else {
for (int i = (int)len-1; i >= 0; -- i) {
if(mms->test(i)) {
nm++;
// There's a mismatch at this position
assert_gt((int)refcs->size(), i);
char refChar = toupper((*refcs)[i]);
ASSERT_ONLY(char qryChar = (h.fw ? (*pat)[i] : (*pat)[len-i-1]));
assert_neq(refChar, qryChar);
ss << run << refChar;
run = 0;
} else {
run++;
}
}
}
ss << run;
// Add optional edit distance field
ss << "\tNM:i:" << nm;
if(h.color) ss << "\tCM:i:" << h.cmms.count();
// Add optional fields reporting the primer base and the downstream color,
// which, if they were present, were clipped when the read was read in
if(h.color && gReportColorPrimer) {
if(h.primer != '?') {
ss << "\tZP:Z:" << h.primer;
assert(isprint(h.primer));
}
if(h.trimc != '?') {
ss << "\tZp:Z:" << h.trimc;
assert(isprint(h.trimc));
}
}
if(xms > 0) ss << "\tXM:i:" << xms;
ss << endl;
}
/**
* Report a verbose, human-readable alignment to the appropriate
* output stream.
*/
void SAMHitSink::reportSamHit(const Hit& h, int mapq, int xms) {
if(xms == 0) {
// Otherwise, this is actually a sampled read and belongs in
// the same category as maxed reads
HitSink::reportHit(h);
}
ostringstream ss;
append(ss, h, mapq, xms);
// Make sure to grab lock before writing to output stream
lock(h.h.first);
out(h.h.first).writeString(ss.str());
unlock(h.h.first);
}
/**
* Report a batch of hits from a vector, perhaps subsetting it.
*/
void SAMHitSink::reportSamHits(
vector<Hit>& hs,
size_t start,
size_t end,
int mapq,
int xms)
{
assert_geq(end, start);
if(end-start == 0) return;
assert_gt(hs[start].mate, 0);
char buf[4096];
lock(0);
for(size_t i = start; i < end; i++) {
ostringstream ss(ssmode_);
ss.rdbuf()->pubsetbuf(buf, 4096);
append(ss, hs[i], mapq, xms);
out(0).writeChars(buf, ss.tellp());
}
unlock(0);
mainlock();
commitHits(hs);
first_ = false;
numAligned_++;
numReportedPaired_ += (end-start);
mainunlock();
}
/**
* Report either an unaligned read or a read that exceeded the -m
* ceiling. We output placeholders for most of the fields in this
* case.
*/
void SAMHitSink::reportUnOrMax(PatternSourcePerThread& p,
vector<Hit>* hs,
bool un) // lower bound on number of other hits
{
if(un) HitSink::reportUnaligned(p);
else HitSink::reportMaxed(*hs, p);
ostringstream ss;
bool paired = !p.bufb().empty();
assert(paired || p.bufa().mate == 0);
assert(!paired || p.bufa().mate > 0);
assert(un || hs->size() > 0);
assert(!un || hs == NULL || hs->size() == 0);
size_t hssz = 0;
if(hs != NULL) hssz = hs->size();
if(paired) {
// truncate final 2 chars
for(int i = 0; i < (int)seqan::length(p.bufa().name)-2; i++) {
if(!noQnameTrunc_ && isspace((int)p.bufa().name[i])) break;
ss << p.bufa().name[i];
}
} else {
for(int i = 0; i < (int)seqan::length(p.bufa().name); i++) {
if(!noQnameTrunc_ && isspace((int)p.bufa().name[i])) break;
ss << p.bufa().name[i];
}
}
ss << "\t"
<< (SAM_FLAG_UNMAPPED | (paired ? (SAM_FLAG_PAIRED | SAM_FLAG_FIRST_IN_PAIR | SAM_FLAG_MATE_UNMAPPED) : 0)) << "\t*"
<< "\t0\t0\t*\t*\t0\t0\t"
<< p.bufa().patFw << "\t" << p.bufa().qual << "\tXM:i:"
<< (paired ? (hssz+1)/2 : hssz);
// Add optional fields reporting the primer base and the downstream color,
// which, if they were present, were clipped when the read was read in
if(p.bufa().color && gReportColorPrimer) {
if(p.bufa().primer != '?') {
ss << "\tZP:Z:" << p.bufa().primer;
assert(isprint(p.bufa().primer));
}
if(p.bufa().trimc != '?') {
ss << "\tZp:Z:" << p.bufa().trimc;
assert(isprint(p.bufa().trimc));
}
}
ss << endl;
if(paired) {
// truncate final 2 chars
for(int i = 0; i < (int)seqan::length(p.bufb().name)-2; i++) {
ss << p.bufb().name[i];
}
ss << "\t"
<< (SAM_FLAG_UNMAPPED | (paired ? (SAM_FLAG_PAIRED | SAM_FLAG_SECOND_IN_PAIR | SAM_FLAG_MATE_UNMAPPED) : 0)) << "\t*"
<< "\t0\t0\t*\t*\t0\t0\t"
<< p.bufb().patFw << "\t" << p.bufb().qual << "\tXM:i:"
<< (hssz+1)/2;
// Add optional fields reporting the primer base and the downstream color,
// which, if they were present, were clipped when the read was read in
if(p.bufb().color && gReportColorPrimer) {
if(p.bufb().primer != '?') {
ss << "\tZP:Z:" << p.bufb().primer;
assert(isprint(p.bufb().primer));
}
if(p.bufb().trimc != '?') {
ss << "\tZp:Z:" << p.bufb().trimc;
assert(isprint(p.bufb().trimc));
}
}
ss << endl;
}
lock(0);
out(0).writeString(ss.str());
unlock(0);
}
/**
* Append a SAM alignment to the given output stream.
*/
void SAMHitSink::append(ostream& ss,
const Hit& h,
int mapq,
int xms,
const vector<string>* refnames,
ReferenceMap *rmap,
AnnotationMap *amap,
bool fullRef,
bool noQnameTrunc,
int offBase)
{
appendAligned(ss, h, mapq, xms, refnames, rmap, amap, fullRef, noQnameTrunc, offBase);
}
/**
* Report maxed-out read; if sampleMax_ is set, then report 1 alignment
* at random.
*/
void SAMHitSink::reportMaxed(vector<Hit>& hs, PatternSourcePerThread& p) {
if(sampleMax_) {
HitSink::reportMaxed(hs, p);
RandomSource rand;
rand.init(p.bufa().seed);
assert_gt(hs.size(), 0);
bool paired = hs.front().mate > 0;
size_t num = 1;
if(paired) {
num = 0;
int bestStratum = 999;
for(size_t i = 0; i < hs.size()-1; i += 2) {
int strat = min(hs[i].stratum, hs[i+1].stratum);
if(strat < bestStratum) {
bestStratum = strat;
num = 1;
} else if(strat == bestStratum) {
num++;
}
}
assert_leq(num, hs.size());
uint32_t r = rand.nextU32() % num;
num = 0;
for(size_t i = 0; i < hs.size()-1; i += 2) {
int strat = min(hs[i].stratum, hs[i+1].stratum);
if(strat == bestStratum) {
if(num == r) {
reportSamHits(hs, i, i+2, 0, (int)(hs.size()/2)+1);
break;
}
num++;
}
}
assert_eq(num, r);
} else {
for(size_t i = 1; i < hs.size(); i++) {
assert_geq(hs[i].stratum, hs[i-1].stratum);
if(hs[i].stratum == hs[i-1].stratum) num++;
else break;
}
assert_leq(num, hs.size());
uint32_t r = rand.nextU32() % num;
reportSamHit(hs[r], /*MAPQ*/0, /*XM:I*/(int)hs.size()+1);
}
} else {
reportUnOrMax(p, &hs, false);
}
}
|