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realn.c
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/* realn.c -- BAQ calculation and realignment.
Copyright (C) 2009-2011, 2014-2015 Genome Research Ltd.
Portions copyright (C) 2009-2011 Broad Institute.
Author: Heng Li <[email protected]>
Permission is hereby granted, free of charge, to any person obtaining a copy
of this software and associated documentation files (the "Software"), to deal
in the Software without restriction, including without limitation the rights
to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
copies of the Software, and to permit persons to whom the Software is
furnished to do so, subject to the following conditions:
The above copyright notice and this permission notice shall be included in
all copies or substantial portions of the Software.
THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL
THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING
FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER
DEALINGS IN THE SOFTWARE. */
#include <config.h>
#include <stdlib.h>
#include <string.h>
#include <math.h>
#include "htslib/hts.h"
#include "htslib/sam.h"
int sam_cap_mapq(bam1_t *b, const char *ref, int ref_len, int thres)
{
uint8_t *seq = bam_get_seq(b), *qual = bam_get_qual(b);
uint32_t *cigar = bam_get_cigar(b);
bam1_core_t *c = &b->core;
int i, x, y, mm, q, len, clip_l, clip_q;
double t;
if (thres < 0) thres = 40; // set the default
mm = q = len = clip_l = clip_q = 0;
for (i = y = 0, x = c->pos; i < c->n_cigar; ++i) {
int j, l = cigar[i]>>4, op = cigar[i]&0xf;
if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) {
for (j = 0; j < l; ++j) {
int c1, c2, z = y + j;
if (x+j >= ref_len || ref[x+j] == '\0') break; // out of bounds
c1 = bam_seqi(seq, z), c2 = seq_nt16_table[(int)ref[x+j]];
if (c2 != 15 && c1 != 15 && qual[z] >= 13) { // not ambiguous
++len;
if (c1 && c1 != c2 && qual[z] >= 13) { // mismatch
++mm;
q += qual[z] > 33? 33 : qual[z];
}
}
}
if (j < l) break;
x += l; y += l; len += l;
} else if (op == BAM_CDEL) {
for (j = 0; j < l; ++j)
if (x+j >= ref_len || ref[x+j] == '\0') break;
if (j < l) break;
x += l;
} else if (op == BAM_CSOFT_CLIP) {
for (j = 0; j < l; ++j) clip_q += qual[y+j];
clip_l += l;
y += l;
} else if (op == BAM_CHARD_CLIP) {
clip_q += 13 * l;
clip_l += l;
} else if (op == BAM_CINS) y += l;
else if (op == BAM_CREF_SKIP) x += l;
}
for (i = 0, t = 1; i < mm; ++i)
t *= (double)len / (i+1);
t = q - 4.343 * log(t) + clip_q / 5.;
if (t > thres) return -1;
if (t < 0) t = 0;
t = sqrt((thres - t) / thres) * thres;
//fprintf(stderr, "%s %lf %d\n", bam_get_qname(b), t, q);
return (int)(t + .499);
}
int sam_prob_realn(bam1_t *b, const char *ref, int ref_len, int flag)
{
int k, i, bw, x, y, yb, ye, xb, xe, apply_baq = flag&1, extend_baq = flag>>1&1, redo_baq = flag&4;
uint32_t *cigar = bam_get_cigar(b);
bam1_core_t *c = &b->core;
probaln_par_t conf = { 0.001, 0.1, 10 };
uint8_t *bq = 0, *zq = 0, *qual = bam_get_qual(b);
if ((c->flag & BAM_FUNMAP) || b->core.l_qseq == 0 || qual[0] == (uint8_t)-1)
return -1; // do nothing
// test if BQ or ZQ is present
if ((bq = bam_aux_get(b, "BQ")) != 0) ++bq;
if ((zq = bam_aux_get(b, "ZQ")) != 0 && *zq == 'Z') ++zq;
if (bq && redo_baq)
{
bam_aux_del(b, bq-1);
bq = 0;
}
if (bq && zq) { // remove the ZQ tag
bam_aux_del(b, zq-1);
zq = 0;
}
if (bq || zq) {
if ((apply_baq && zq) || (!apply_baq && bq)) return -3; // in both cases, do nothing
if (bq && apply_baq) { // then convert BQ to ZQ
for (i = 0; i < c->l_qseq; ++i)
qual[i] = qual[i] + 64 < bq[i]? 0 : qual[i] - ((int)bq[i] - 64);
*(bq - 3) = 'Z';
} else if (zq && !apply_baq) { // then convert ZQ to BQ
for (i = 0; i < c->l_qseq; ++i)
qual[i] += (int)zq[i] - 64;
*(zq - 3) = 'B';
}
return 0;
}
// find the start and end of the alignment
x = c->pos, y = 0, yb = ye = xb = xe = -1;
for (k = 0; k < c->n_cigar; ++k) {
int op, l;
op = cigar[k]&0xf; l = cigar[k]>>4;
if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) {
if (yb < 0) yb = y;
if (xb < 0) xb = x;
ye = y + l; xe = x + l;
x += l; y += l;
} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
else if (op == BAM_CDEL) x += l;
else if (op == BAM_CREF_SKIP) return -1; // do nothing if there is a reference skip
}
// set bandwidth and the start and the end
bw = 7;
if (abs((xe - xb) - (ye - yb)) > bw)
bw = abs((xe - xb) - (ye - yb)) + 3;
conf.bw = bw;
xb -= yb + bw/2; if (xb < 0) xb = 0;
xe += c->l_qseq - ye + bw/2;
if (xe - xb - c->l_qseq > bw)
xb += (xe - xb - c->l_qseq - bw) / 2, xe -= (xe - xb - c->l_qseq - bw) / 2;
{ // glocal
uint8_t *s, *r, *q, *seq = bam_get_seq(b), *bq;
int *state;
bq = calloc(c->l_qseq + 1, 1);
memcpy(bq, qual, c->l_qseq);
s = calloc(c->l_qseq, 1);
for (i = 0; i < c->l_qseq; ++i) s[i] = seq_nt16_int[bam_seqi(seq, i)];
r = calloc(xe - xb, 1);
for (i = xb; i < xe; ++i) {
if (i >= ref_len || ref[i] == '\0') { xe = i; break; }
r[i-xb] = seq_nt16_int[seq_nt16_table[(int)ref[i]]];
}
state = calloc(c->l_qseq, sizeof(int));
q = calloc(c->l_qseq, 1);
probaln_glocal(r, xe-xb, s, c->l_qseq, qual, &conf, state, q);
if (!extend_baq) { // in this block, bq[] is capped by base quality qual[]
for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) {
int op = cigar[k]&0xf, l = cigar[k]>>4;
if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) {
for (i = y; i < y + l; ++i) {
if ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y)) bq[i] = 0;
else bq[i] = bq[i] < q[i]? bq[i] : q[i];
}
x += l; y += l;
} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
else if (op == BAM_CDEL) x += l;
}
for (i = 0; i < c->l_qseq; ++i) bq[i] = qual[i] - bq[i] + 64; // finalize BQ
} else { // in this block, bq[] is BAQ that can be larger than qual[] (different from the above!)
uint8_t *left, *rght;
left = calloc(c->l_qseq, 1); rght = calloc(c->l_qseq, 1);
for (k = 0, x = c->pos, y = 0; k < c->n_cigar; ++k) {
int op = cigar[k]&0xf, l = cigar[k]>>4;
if (op == BAM_CMATCH || op == BAM_CEQUAL || op == BAM_CDIFF) {
for (i = y; i < y + l; ++i)
bq[i] = ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y))? 0 : q[i];
for (left[y] = bq[y], i = y + 1; i < y + l; ++i)
left[i] = bq[i] > left[i-1]? bq[i] : left[i-1];
for (rght[y+l-1] = bq[y+l-1], i = y + l - 2; i >= y; --i)
rght[i] = bq[i] > rght[i+1]? bq[i] : rght[i+1];
for (i = y; i < y + l; ++i)
bq[i] = left[i] < rght[i]? left[i] : rght[i];
x += l; y += l;
} else if (op == BAM_CSOFT_CLIP || op == BAM_CINS) y += l;
else if (op == BAM_CDEL) x += l;
}
for (i = 0; i < c->l_qseq; ++i) bq[i] = 64 + (qual[i] <= bq[i]? 0 : qual[i] - bq[i]); // finalize BQ
free(left); free(rght);
}
if (apply_baq) {
for (i = 0; i < c->l_qseq; ++i) qual[i] -= bq[i] - 64; // modify qual
bam_aux_append(b, "ZQ", 'Z', c->l_qseq + 1, bq);
} else bam_aux_append(b, "BQ", 'Z', c->l_qseq + 1, bq);
free(bq); free(s); free(r); free(q); free(state);
}
return 0;
}