#include #include #include #include #include "faidx.h" #include "sam.h" #include "kstring.h" void bam_fillmd1_core(bam1_t *b, char *ref, int is_equal, int max_nm) { uint8_t *seq = bam1_seq(b); uint32_t *cigar = bam1_cigar(b); bam1_core_t *c = &b->core; int i, x, y, u = 0; kstring_t *str; uint8_t *old_md, *old_nm; int32_t old_nm_i = -1, nm = 0; str = (kstring_t*)calloc(1, sizeof(kstring_t)); 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 ((c1 == c2 && c1 != 15 && c2 != 15) || c1 == 0) { // a match if (is_equal) seq[z/2] &= (z&1)? 0xf0 : 0x0f; ++u; } else { ksprintf(str, "%d", u); kputc(ref[x+j], str); u = 0; ++nm; } } if (j < l) break; x += l; y += l; } else if (op == BAM_CDEL) { ksprintf(str, "%d", u); kputc('^', str); for (j = 0; j < l; ++j) { if (ref[x+j] == 0) break; kputc(ref[x+j], str); } u = 0; if (j < l) break; x += l; nm += l; } else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) { y += l; if (op == BAM_CINS) nm += l; } else if (op == BAM_CREF_SKIP) { x += l; } } ksprintf(str, "%d", u); // apply max_nm if (max_nm > 0 && nm >= max_nm) { 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 ((c1 == c2 && c1 != 15 && c2 != 15) || c1 == 0) { // a match seq[z/2] |= (z&1)? 0x0f : 0xf0; bam1_qual(b)[z] = 0; } } if (j < l) break; x += l; y += l; } else if (op == BAM_CDEL || op == BAM_CREF_SKIP) x += l; else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) y += l; } } // update NM old_nm = bam_aux_get(b, "NM"); if (c->flag & BAM_FUNMAP) return; if (old_nm) old_nm_i = bam_aux2i(old_nm); if (!old_nm) bam_aux_append(b, "NM", 'i', 4, (uint8_t*)&nm); else if (nm != old_nm_i) { fprintf(stderr, "[bam_fillmd1] different NM for read '%s': %d -> %d\n", bam1_qname(b), old_nm_i, nm); bam_aux_del(b, old_nm); bam_aux_append(b, "NM", 'i', 4, (uint8_t*)&nm); } // update MD old_md = bam_aux_get(b, "MD"); if (!old_md) bam_aux_append(b, "MD", 'Z', str->l + 1, (uint8_t*)str->s); else { int is_diff = 0; if (strlen((char*)old_md+1) == str->l) { for (i = 0; i < str->l; ++i) if (toupper(old_md[i+1]) != toupper(str->s[i])) break; if (i < str->l) is_diff = 1; } else is_diff = 1; if (is_diff) { fprintf(stderr, "[bam_fillmd1] different MD for read '%s': '%s' -> '%s'\n", bam1_qname(b), old_md+1, str->s); bam_aux_del(b, old_md); bam_aux_append(b, "MD", 'Z', str->l + 1, (uint8_t*)str->s); } } free(str->s); free(str); } void bam_fillmd1(bam1_t *b, char *ref, int is_equal) { bam_fillmd1_core(b, ref, is_equal, 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; samfile_t *fp, *fpout = 0; faidx_t *fai; char *ref = 0, mode_w[8], mode_r[8]; bam1_t *b; is_bam_out = is_sam_in = is_uncompressed = 0; mode_w[0] = mode_r[0] = 0; strcpy(mode_r, "r"); strcpy(mode_w, "w"); while ((c = getopt(argc, argv, "eubSn:")) >= 0) { switch (c) { case 'e': is_equal = 1; break; case 'b': is_bam_out = 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; default: fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n", c); return 1; } } if (!is_sam_in) strcat(mode_r, "b"); if (is_bam_out) strcat(mode_w, "b"); else strcat(mode_w, "h"); if (is_uncompressed) strcat(mode_w, "u"); if (optind + 1 >= argc) { fprintf(stderr, "\n"); fprintf(stderr, "Usage: samtools fillmd [-eubS] \n\n"); fprintf(stderr, "Options: -e change identical bases to '='\n"); fprintf(stderr, " -u uncompressed BAM output (for piping)\n"); fprintf(stderr, " -b compressed BAM output\n"); fprintf(stderr, " -S the input is SAM with header\n\n"); return 1; } fp = samopen(argv[optind], mode_r, 0); if (fp == 0) return 1; if (is_sam_in && (fp->header == 0 || fp->header->n_targets == 0)) { fprintf(stderr, "[bam_fillmd] input SAM does not have header. Abort!\n"); return 1; } fpout = samopen("-", mode_w, fp->header); fai = fai_load(argv[optind+1]); b = bam_init1(); while ((ret = samread(fp, b)) >= 0) { if (b->core.tid >= 0) { if (tid != b->core.tid) { free(ref); ref = fai_fetch(fai, fp->header->target_name[b->core.tid], &len); tid = b->core.tid; if (ref == 0) fprintf(stderr, "[bam_fillmd] fail to find sequence '%s' in the reference.\n", fp->header->target_name[tid]); } if (ref) bam_fillmd1_core(b, ref, is_equal, max_nm); } samwrite(fpout, b); } bam_destroy1(b); free(ref); fai_destroy(fai); samclose(fp); samclose(fpout); return 0; }