62
62
long double sum_a[257], b[256], q_c[256], tmp[256], fk2[256];
65
lC = (double*)calloc(256 * 256, sizeof(double));
66
65
// aa->lhet will be allocated and initialized
67
66
free(aa->fk); free(aa->coef);
68
68
aa->fk = (double*)calloc(256, sizeof(double));
69
aa->coef = (double*)calloc(256*256*64, sizeof(double));
70
69
aa->fk[0] = fk2[0] = 1.0;
71
70
for (n = 1; n != 256; ++n) {
72
71
aa->fk[n] = pow(aa->theta, n) * (1.0 - aa->eta) + aa->eta;
73
72
fk2[n] = aa->fk[n>>1]; // this is an approximation, assuming reads equally likely come from both strands
74
if (aa->is_soap) return;
75
aa->coef = (double*)calloc(256*256*64, sizeof(double));
76
lC = (double*)calloc(256 * 256, sizeof(double));
75
77
for (n = 1; n != 256; ++n)
76
78
for (k = 1; k <= n; ++k)
77
79
lC[n<<8|k] = lgamma(n+1) - lgamma(k+1) - lgamma(n-k+1);
180
182
for (j = 0; j != 4; ++j) b->c[j] = (int)(254.0 * b->c[j] / c + 0.5);
181
183
for (j = c = 0; j != 4; ++j) c += b->c[j];
183
// generate likelihood
184
for (j = 0; j != 4; ++j) {
188
for (k = 0, tmp1 = tmp3 = 0.0, tmp2 = 0; k != 4; ++k) {
189
if (j == k) continue;
190
tmp1 += b->esum[k]; tmp2 += b->c[k]; tmp3 += b->fsum[k];
193
bar_e = (int)(tmp1 / tmp3 + 0.5);
194
if (bar_e < 4) bar_e = 4; // should not happen
195
if (bar_e > 63) bar_e = 63;
196
p[j<<2|j] = tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
197
} else p[j<<2|j] = 0.0; // all the bases are j
199
for (k = j + 1; k < 4; ++k) {
200
for (i = 0, tmp2 = 0, tmp1 = tmp3 = 0.0; i != 4; ++i) {
201
if (i == j || i == k) continue;
202
tmp1 += b->esum[i]; tmp2 += b->c[i]; tmp3 += b->fsum[i];
186
// generate likelihood
187
for (j = 0; j != 4; ++j) {
191
for (k = 0, tmp1 = tmp3 = 0.0, tmp2 = 0; k != 4; ++k) {
192
if (j == k) continue;
193
tmp1 += b->esum[k]; tmp2 += b->c[k]; tmp3 += b->fsum[k];
205
196
bar_e = (int)(tmp1 / tmp3 + 0.5);
206
if (bar_e < 4) bar_e = 4;
197
if (bar_e < 4) bar_e = 4; // should not happen
207
198
if (bar_e > 63) bar_e = 63;
208
p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
209
} else p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]]; // all the bases are either j or k
199
p[j<<2|j] = tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
200
} else p[j<<2|j] = 0.0; // all the bases are j
202
for (k = j + 1; k < 4; ++k) {
203
for (i = 0, tmp2 = 0, tmp1 = tmp3 = 0.0; i != 4; ++i) {
204
if (i == j || i == k) continue;
205
tmp1 += b->esum[i]; tmp2 += b->c[i]; tmp3 += b->fsum[i];
208
bar_e = (int)(tmp1 / tmp3 + 0.5);
209
if (bar_e < 4) bar_e = 4;
210
if (bar_e > 63) bar_e = 63;
211
p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp1 + bm->coef[bar_e<<16|c<<8|tmp2];
212
} else p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]]; // all the bases are either j or k
215
for (k = 0; k != 4; ++k)
216
if (p[j<<2|k] < 0.0) p[j<<2|k] = 0.0;
212
for (k = 0; k != 4; ++k)
213
if (p[j<<2|k] < 0.0) p[j<<2|k] = 0.0;
217
float max1, max2, min1, min2;
220
max1 = max2 = -1.0; min1 = min2 = 1e30;
221
for (k = 0; k < 4; ++k) {
222
if (b->esum[k] > max1) {
223
max2 = max1; max1 = b->esum[k]; max_k = k;
224
} else if (b->esum[k] > max2) max2 = b->esum[k];
226
for (k = 0; k < 4; ++k) {
227
if (p[k<<2|k] < min1) {
228
min2 = min1; min1 = p[k<<2|k]; min_k = k;
229
} else if (p[k<<2|k] < min2) min2 = p[k<<2|k];
231
if (max1 > max2 && (min_k != max_k || min1 + 1.0 > min2))
232
p[max_k<<2|max_k] = min1 > 1.0? min1 - 1.0 : 0.0;
220
float max1, max2, min1, min2;
223
max1 = max2 = -1.0; min1 = min2 = 1e30;
224
for (k = 0; k < 4; ++k) {
225
if (b->esum[k] > max1) {
226
max2 = max1; max1 = b->esum[k]; max_k = k;
227
} else if (b->esum[k] > max2) max2 = b->esum[k];
229
for (k = 0; k < 4; ++k) {
230
if (p[k<<2|k] < min1) {
231
min2 = min1; min1 = p[k<<2|k]; min_k = k;
232
} else if (p[k<<2|k] < min2) min2 = p[k<<2|k];
234
if (max1 > max2 && (min_k != max_k || min1 + 1.0 > min2))
235
p[max_k<<2|max_k] = min1 > 1.0? min1 - 1.0 : 0.0;
237
} else { // apply the SOAP model
238
// generate likelihood
239
for (j = 0; j != 4; ++j) {
242
for (k = 0, tmp = 0.0; k != 4; ++k)
243
if (j != k) tmp += b->esum[k];
246
for (k = j + 1; k < 4; ++k) {
247
for (i = 0, tmp = 0.0; i != 4; ++i)
248
if (i != j && i != k) tmp += b->esum[i];
249
p[j<<2|k] = p[k<<2|j] = -4.343 * bm->lhet[b->c[j]<<8|b->c[k]] + tmp;
235
254
// convert necessary information to glf1_t
304
323
free(mir->s[0]); free(mir->s[1]); free(mir);
326
int bam_tpos2qpos(const bam1_core_t *c, const uint32_t *cigar, int32_t tpos, int is_left, int32_t *_tpos)
328
int k, x = c->pos, y = 0, last_y = 0;
330
for (k = 0; k < c->n_cigar; ++k) {
331
int op = cigar[k] & BAM_CIGAR_MASK;
332
int l = cigar[k] >> BAM_CIGAR_SHIFT;
333
if (op == BAM_CMATCH) {
334
if (c->pos > tpos) return y;
337
return y + (tpos - x);
341
} else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) y += l;
342
else if (op == BAM_CDEL || op == BAM_CREF_SKIP) {
344
*_tpos = is_left? x : x + l;
307
354
#define MINUS_CONST 0x10000000
309
356
bam_maqindel_ret_t *bam_maqindel(int n, int pos, const bam_maqindel_opt_t *mi, const bam_pileup1_t *pl, const char *ref,
310
357
int _n_types, int *_types)
312
int i, j, n_types, *types, left, right;
359
int i, j, n_types, *types, left, right, max_rd_len = 0;
313
360
bam_maqindel_ret_t *ret = 0;
314
361
// if there is no proposed indel, check if there is an indel from the alignment
315
362
if (_n_types == 0) {
349
398
{ // calculate left and right boundary
351
left = 0x7fffffff; right = 0;
352
for (i = 0; i < n; ++i) {
353
const bam_pileup1_t *p = pl + i;
354
if (!(p->b->core.flag&BAM_FUNMAP)) {
355
bam_segreg(pos, &p->b->core, bam1_cigar(p->b), &seg);
356
if (seg.tbeg < left) left = seg.tbeg;
357
if (seg.tend > right) right = seg.tend;
360
if (pos - left > MAX_WINDOW) left = pos - MAX_WINDOW;
361
if (right - pos> MAX_WINDOW) right = pos + MAX_WINDOW;
399
left = pos > INDEL_WINDOW_SIZE? pos - INDEL_WINDOW_SIZE : 0;
400
right = pos + INDEL_WINDOW_SIZE;
401
if (types[0] < 0) right -= types[0];
402
// in case the alignments stand out the reference
403
for (i = pos; i < right; ++i)
404
if (ref[i] == 0) break;
363
407
{ // the core part
364
char *ref2, *inscns = 0;
408
char *ref2, *rs, *inscns = 0;
365
409
int k, l, *score, *pscore, max_ins = types[n_types-1];
366
ref2 = (char*)calloc(right - left + types[n_types-1] + 2, 1);
367
410
if (max_ins > 0) { // get the consensus of inserted sequences
368
411
int *inscns_aux = (int*)calloc(4 * n_types * max_ins, sizeof(int));
369
412
// count occurrences
396
439
free(inscns_aux);
398
441
// calculate score
442
ref2 = (char*)calloc(right - left + types[n_types-1] + 2, 1);
443
rs = (char*)calloc(right - left + max_rd_len + types[n_types-1] + 2, 1);
399
444
score = (int*)calloc(n_types * n, sizeof(int));
400
445
pscore = (int*)calloc(n_types * n, sizeof(int));
401
446
for (i = 0; i < n_types; ++i) {
447
ka_param_t ap = ka_param_blast;
448
ap.band_width = 2 * types[n_types - 1] + 2;
403
450
for (k = 0, j = left; j <= pos; ++j)
404
ref2[k++] = bam_nt16_table[(int)ref[j]];
451
ref2[k++] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[j]]];
405
452
if (types[i] <= 0) j += -types[i];
406
453
else for (l = 0; l < types[i]; ++l)
407
ref2[k++] = inscns[i*max_ins + l];
454
ref2[k++] = bam_nt16_nt4_table[(int)inscns[i*max_ins + l]];
408
455
for (; j < right && ref[j]; ++j)
409
ref2[k++] = bam_nt16_table[(int)ref[j]];
456
ref2[k++] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[j]]];
457
if (j < right) right = j;
410
458
// calculate score for each read
411
459
for (j = 0; j < n; ++j) {
412
460
const bam_pileup1_t *p = pl + j;
414
bam1_core_t *c = &p->b->core;
417
if (c->flag&BAM_FUNMAP) continue;
418
cigar = bam1_cigar(p->b);
419
bam_segreg(pos, c, cigar, &seg);
420
for (ps = s = 0, l = seg.qbeg; c->pos + l < right && l < seg.qend; ++l) {
421
int cq = bam1_seqi(bam1_seq(p->b), l), ct;
422
// in the following line, "<" will happen if reads are too long
423
ct = c->pos + l - seg.qbeg >= left? ref2[c->pos + l - seg.qbeg - left] : 15;
424
if (cq < 15 && ct < 15) {
425
s += cq == ct? 1 : -mi->mm_penalty;
426
if (cq != ct) ps += bam1_qual(p->b)[l];
461
int qbeg, qend, tbeg, tend;
462
if (p->b->core.flag & BAM_FUNMAP) continue;
463
qbeg = bam_tpos2qpos(&p->b->core, bam1_cigar(p->b), left, 0, &tbeg);
464
qend = bam_tpos2qpos(&p->b->core, bam1_cigar(p->b), right, 1, &tend);
465
assert(tbeg >= left);
466
for (l = qbeg; l < qend; ++l)
467
rs[l - qbeg] = bam_nt16_nt4_table[bam1_seqi(bam1_seq(p->b), l)];
469
int x, y, n_acigar, ps;
472
if (tend - tbeg + types[i] <= 0) {
473
score[i*n+j] = -(1<<20);
474
pscore[i*n+j] = 1<<20;
429
score[i*n + j] = s; pscore[i*n + j] = ps;
430
if (types[i] != 0) { // then try the other way to calculate the score
431
for (ps = s = 0, l = seg.qbeg; c->pos + l + types[i] < right && l < seg.qend; ++l) {
432
int cq = bam1_seqi(bam1_seq(p->b), l), ct;
433
ct = c->pos + l - seg.qbeg + types[i] >= left? ref2[c->pos + l - seg.qbeg + types[i] - left] : 15;
434
if (cq < 15 && ct < 15) {
435
s += cq == ct? 1 : -mi->mm_penalty;
436
if (cq != ct) ps += bam1_qual(p->b)[l];
477
acigar = ka_global_core((uint8_t*)ref2 + tbeg - left, tend - tbeg + types[i], (uint8_t*)rs, qend - qbeg, &ap, &score[i*n+j], &n_acigar);
478
x = tbeg - left; y = 0;
479
for (l = 0; l < n_acigar; ++l) {
480
int op = acigar[l]&0xf;
481
int len = acigar[l]>>4;
482
if (op == BAM_CMATCH) {
484
for (k = 0; k < len; ++k)
485
if (ref2[x+k] != rs[y+k]) ps += bam1_qual(p->b)[y+k];
487
} else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) {
488
if (op == BAM_CINS) ps += mi->q_indel * len;
490
} else if (op == BAM_CDEL) {
491
ps += mi->q_indel * len;
496
/*if (pos == 2618517) { // for debugging only
497
fprintf(stderr, "pos=%d, type=%d, j=%d, score=%d, psore=%d, %d, %d, %d, %d, ", pos+1, types[i], j, score[i*n+j], pscore[i*n+j], tbeg, tend, qbeg, qend);
498
for (l = 0; l < n_acigar; ++l) fprintf(stderr, "%d%c", acigar[l]>>4, "MIDS"[acigar[l]&0xf]); fprintf(stderr, "\n");
499
for (l = 0; l < tend - tbeg + types[i]; ++l) fputc("ACGTN"[ref2[l]], stderr); fputc('\n', stderr);
500
for (l = 0; l < qend - qbeg; ++l) fputc("ACGTN"[rs[l]], stderr); fputc('\n', stderr);
440
if (score[i*n+j] < s) score[i*n+j] = s; // choose the higher of the two scores
441
if (pscore[i*n+j] > ps) pscore[i*n+j] = ps;
442
//if (types[i] != 0) score[i*n+j] -= mi->indel_err;
443
//printf("%d, %d, %d, %d, %d, %d, %d\n", p->b->core.pos + 1, seg.qbeg, i, types[i], j,
444
// score[i*n+j], pscore[i*n+j]);
447
506
{ // get final result
491
550
else if (p->indel == ret->indel2) ++ret->cnt2;
492
551
else ++ret->cnt_anti;
495
ret->gl[0] = ret->gl[1] = 0;
496
for (j = 0; j < n; ++j) {
497
int s1 = pscore[max1_i*n + j], s2 = pscore[max2_i*n + j];
498
//printf("%d, %d, %d, %d, %d\n", pl[j].b->core.pos+1, max1_i, max2_i, s1, s2);
499
if (s1 > s2) ret->gl[0] += s1 - s2 < mi->q_indel? s1 - s2 : mi->q_indel;
500
else ret->gl[1] += s2 - s1 < mi->q_indel? s2 - s1 : mi->q_indel;
554
int tmp, seq_err = 0;
556
tmp = max1_i - max2_i;
557
if (tmp < 0) tmp = -tmp;
558
for (j = 0; j < tmp + 1; ++j) x *= INDEL_EXT_DEP;
559
seq_err = mi->q_indel * (1.0 - x) / (1.0 - INDEL_EXT_DEP);
560
ret->gl[0] = ret->gl[1] = 0;
561
for (j = 0; j < n; ++j) {
562
int s1 = pscore[max1_i*n + j], s2 = pscore[max2_i*n + j];
563
//printf("%d, %d, %d, %d, %d\n", pl[j].b->core.pos+1, max1_i, max2_i, s1, s2);
564
if (s1 > s2) ret->gl[0] += s1 - s2 < seq_err? s1 - s2 : seq_err;
565
else ret->gl[1] += s2 - s1 < seq_err? s2 - s1 : seq_err;
502
568
// write cnt_ref and cnt_ambi
503
569
if (max1_i != 0 && max2_i != 0) {