#include <assert.h>
#include <string.h>
#include <ctype.h>
+#include <math.h>
#include "faidx.h"
#include "sam.h"
#include "kstring.h"
#include "kaln.h"
+#include "kprobaln.h"
void bam_fillmd1_core(bam1_t *b, char *ref, int is_equal, int max_nm)
{
bam_fillmd1_core(b, ref, is_equal, 0);
}
-// local realignment
-
-#define MIN_REF_LEN 10
-#define MIN_BAND_WIDTH 11
-
-int bam_realn(bam1_t *b, const char *ref)
+int bam_cap_mapQ(bam1_t *b, char *ref, int thres)
{
- int k, l_ref, score, n_cigar;
+ uint8_t *seq = bam1_seq(b), *qual = bam1_qual(b);
uint32_t *cigar = bam1_cigar(b);
- uint8_t *s_ref = 0, *s_read = 0, *seq;
- ka_param_t par;
bam1_core_t *c = &b->core;
- // set S/W parameters
- par = ka_param_blast;
- par.gap_open = 4; par.gap_ext = 1; par.gap_end_open = par.gap_end_ext = 0;
- if (c->n_cigar > 1) { // set band width
- int sumi, sumd;
- sumi = sumd = 0;
- for (k = 0; k < c->n_cigar; ++k)
- if ((cigar[k]&0xf) == BAM_CINS) sumi += cigar[k]>>4;
- else if ((cigar[k]&0xf) == BAM_CDEL) sumd += cigar[k]>>4;
- par.band_width = (sumi > sumd? sumi : sumd) + MIN_BAND_WIDTH;
- } else par.band_width = MIN_BAND_WIDTH;
- // calculate the length of the reference in the alignment
- for (k = l_ref = 0; k < c->n_cigar; ++k) {
- if ((cigar[k]&0xf) == BAM_CREF_SKIP) break; // do not do realignment if there is an `N' operation
- if ((cigar[k]&0xf) == BAM_CMATCH || (cigar[k]&0xf) == BAM_CDEL)
- l_ref += cigar[k]>>4;
- }
- if (k != c->n_cigar || l_ref < MIN_REF_LEN) return -1;
- for (k = 0; k < l_ref; ++k)
- if (ref[c->pos + k] == 0) return -1; // the read stands out of the reference
- // allocate
- s_ref = calloc(l_ref, 1);
- s_read = calloc(c->l_qseq, 1);
- for (k = 0, seq = bam1_seq(b); k < c->l_qseq; ++k)
- s_read[k] = bam_nt16_nt4_table[bam1_seqi(seq, k)];
- for (k = 0; k < l_ref; ++k)
- s_ref[k] = bam_nt16_nt4_table[(int)bam_nt16_table[(int)ref[c->pos + k]]];
- // do alignment
- cigar = ka_global_core(s_ref, l_ref, s_read, c->l_qseq, &par, &score, &n_cigar);
- if (score <= 0) { // realignment failed
- free(cigar); free(s_ref); free(s_read);
- return -1;
+ 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) {
+ for (j = 0; j < l; ++j) {
+ int z = y + j;
+ int c1 = bam1_seqi(seq, z), c2 = bam_nt16_table[(int)ref[x+j]];
+ if (ref[x+j] == 0) break; // out of boundary
+ 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 (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;
}
- // copy over the alignment
- if (4 * (n_cigar - (int)c->n_cigar) + b->data_len > b->m_data) { // enlarge b->data
- b->m_data = 4 * (n_cigar - (int)c->n_cigar) + b->data_len;
- kroundup32(b->m_data);
- b->data = realloc(b->data, b->m_data);
+ 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", bam1_qname(b), t, q);
+ return (int)(t + .499);
+}
+
+int bam_prob_realn(bam1_t *b, const char *ref)
+{
+ int k, i, bw, x, y, yb, ye, xb, xe;
+ uint32_t *cigar = bam1_cigar(b);
+ bam1_core_t *c = &b->core;
+ kpa_par_t conf = kpa_par_def;
+ // find the start and end of the alignment
+ if (c->flag & BAM_FUNMAP) return -1;
+ 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) {
+ 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;
}
- if (n_cigar != (int)c->n_cigar) { // move data
- memmove(b->data + c->l_qname + 4 * n_cigar, bam1_seq(b), b->data_len - c->l_qname - 4 * c->n_cigar);
- b->data_len += 4 * (n_cigar - (int)c->n_cigar);
+ // 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 = bam1_seq(b), *qual = bam1_qual(b);
+ int *state;
+ s = calloc(c->l_qseq, 1);
+ for (i = 0; i < c->l_qseq; ++i) s[i] = bam_nt16_nt4_table[bam1_seqi(seq, i)];
+ r = calloc(xe - xb, 1);
+ for (i = xb; i < xe; ++i)
+ r[i-xb] = bam_nt16_nt4_table[bam_nt16_table[(int)ref[i]]];
+ state = calloc(c->l_qseq, sizeof(int));
+ q = calloc(c->l_qseq, 1);
+ kpa_glocal(r, xe-xb, s, c->l_qseq, qual, &conf, state, q);
+ 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) {
+ for (i = y; i < y + l; ++i) {
+ if ((state[i]&3) != 0 || state[i]>>2 != x - xb + (i - y)) qual[i] = 0;
+ else qual[i] = qual[i] < q[i]? qual[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;
+ }
+ free(s); free(r); free(q); free(state);
}
- memcpy(bam1_cigar(b), cigar, n_cigar * 4);
- c->n_cigar = n_cigar;
- free(s_ref); free(s_read); free(cigar);
return 0;
}
int bam_fillmd(int argc, char *argv[])
{
- int c, is_equal = 0, tid = -2, ret, len, is_bam_out, is_sam_in, is_uncompressed, max_nm = 0, is_realn;
+ int c, is_equal = 0, tid = -2, ret, len, is_bam_out, is_sam_in, is_uncompressed, max_nm = 0, is_realn, capQ = 0;
samfile_t *fp, *fpout = 0;
faidx_t *fai;
char *ref = 0, mode_w[8], mode_r[8];
is_bam_out = is_sam_in = is_uncompressed = is_realn = 0;
mode_w[0] = mode_r[0] = 0;
strcpy(mode_r, "r"); strcpy(mode_w, "w");
- while ((c = getopt(argc, argv, "reubSn:")) >= 0) {
+ while ((c = getopt(argc, argv, "reubSC:n:")) >= 0) {
switch (c) {
case 'r': is_realn = 1; break;
case 'e': is_equal = 1; break;
case 'u': is_uncompressed = is_bam_out = 1; break;
case 'S': is_sam_in = 1; break;
case 'n': max_nm = atoi(optarg); break;
+ case 'C': capQ = atoi(optarg); break;
default: fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n", c); return 1;
}
}
fprintf(stderr, "[bam_fillmd] fail to find sequence '%s' in the reference.\n",
fp->header->target_name[tid]);
}
- if (is_realn) bam_realn(b, ref);
+ if (is_realn) bam_prob_realn(b, ref);
+ if (capQ > 10) {
+ int q = bam_cap_mapQ(b, ref, capQ);
+ if (b->core.qual > q) b->core.qual = q;
+ }
if (ref) bam_fillmd1_core(b, ref, is_equal, max_nm);
}
samwrite(fpout, b);
projects / samtools.git / blobdiff