/* This program demonstrates how to generate pileup from multiple BAMs * simutaneously, to achieve random access and to use the BED interface. * To compile this program separately, you may: * * gcc -g -O2 -Wall -o bam2depth -D_MAIN_BAM2DEPTH bam2depth.c -L. -lbam -lz */ #include #include #include #include #include "bam.h" typedef struct { // auxiliary data structure bamFile fp; // the file handler bam_iter_t iter; // NULL if a region not specified int min_mapQ, min_len; // mapQ filter; length filter } aux_t; void *bed_read(const char *fn); // read a BED or position list file void bed_destroy(void *_h); // destroy the BED data structure int bed_overlap(const void *_h, const char *chr, int beg, int end); // test if chr:beg-end overlaps // This function reads a BAM alignment from one BAM file. static int read_bam(void *data, bam1_t *b) // read level filters better go here to avoid pileup { aux_t *aux = (aux_t*)data; // data in fact is a pointer to an auxiliary structure int ret = aux->iter? bam_iter_read(aux->fp, aux->iter, b) : bam_read1(aux->fp, b); if (!(b->core.flag&BAM_FUNMAP)) { if ((int)b->core.qual < aux->min_mapQ) b->core.flag |= BAM_FUNMAP; else if (aux->min_len && bam_cigar2qlen(&b->core, bam1_cigar(b)) < aux->min_len) b->core.flag |= BAM_FUNMAP; } return ret; } int read_file_list(const char *file_list,int *n,char **argv[]); #ifdef _MAIN_BAM2DEPTH int main(int argc, char *argv[]) #else int main_depth(int argc, char *argv[]) #endif { int i, n, tid, beg, end, pos, *n_plp, baseQ = 0, mapQ = 0, min_len = 0, nfiles; const bam_pileup1_t **plp; char *reg = 0; // specified region void *bed = 0; // BED data structure char *file_list = NULL, **fn = NULL; bam_header_t *h = 0; // BAM header of the 1st input aux_t **data; bam_mplp_t mplp; // parse the command line while ((n = getopt(argc, argv, "r:b:q:Q:l:f:")) >= 0) { switch (n) { case 'l': min_len = atoi(optarg); break; // minimum query length case 'r': reg = strdup(optarg); break; // parsing a region requires a BAM header case 'b': bed = bed_read(optarg); break; // BED or position list file can be parsed now case 'q': baseQ = atoi(optarg); break; // base quality threshold case 'Q': mapQ = atoi(optarg); break; // mapping quality threshold case 'f': file_list = optarg; break; } } if (optind == argc && !file_list) { fprintf(stderr, "\n"); fprintf(stderr, "Usage: samtools depth [options] in1.bam [in2.bam [...]]\n"); fprintf(stderr, "Options:\n"); fprintf(stderr, " -b list of positions or regions\n"); fprintf(stderr, " -f list of input BAM filenames, one per line [null]\n"); fprintf(stderr, " -l minQLen\n"); fprintf(stderr, " -q base quality threshold\n"); fprintf(stderr, " -Q mapping quality threshold\n"); fprintf(stderr, " -r region\n"); fprintf(stderr, "\n"); return 1; } // initialize the auxiliary data structures if (file_list) { if ( read_file_list(file_list,&nfiles,&fn) ) return 1; n = nfiles; argv = fn; optind = 0; } else n = argc - optind; // the number of BAMs on the command line data = calloc(n, sizeof(void*)); // data[i] for the i-th input beg = 0; end = 1<<30; tid = -1; // set the default region for (i = 0; i < n; ++i) { bam_header_t *htmp; data[i] = calloc(1, sizeof(aux_t)); data[i]->fp = bam_open(argv[optind+i], "r"); // open BAM data[i]->min_mapQ = mapQ; // set the mapQ filter data[i]->min_len = min_len; // set the qlen filter htmp = bam_header_read(data[i]->fp); // read the BAM header if (i == 0) { h = htmp; // keep the header of the 1st BAM if (reg) bam_parse_region(h, reg, &tid, &beg, &end); // also parse the region } else bam_header_destroy(htmp); // if not the 1st BAM, trash the header if (tid >= 0) { // if a region is specified and parsed successfully bam_index_t *idx = bam_index_load(argv[optind+i]); // load the index data[i]->iter = bam_iter_query(idx, tid, beg, end); // set the iterator bam_index_destroy(idx); // the index is not needed any more; phase out of the memory } } // the core multi-pileup loop mplp = bam_mplp_init(n, read_bam, (void**)data); // initialization n_plp = calloc(n, sizeof(int)); // n_plp[i] is the number of covering reads from the i-th BAM plp = calloc(n, sizeof(void*)); // plp[i] points to the array of covering reads (internal in mplp) while (bam_mplp_auto(mplp, &tid, &pos, n_plp, plp) > 0) { // come to the next covered position if (pos < beg || pos >= end) continue; // out of range; skip if (bed && bed_overlap(bed, h->target_name[tid], pos, pos + 1) == 0) continue; // not in BED; skip fputs(h->target_name[tid], stdout); printf("\t%d", pos+1); // a customized printf() would be faster for (i = 0; i < n; ++i) { // base level filters have to go here int j, m = 0; for (j = 0; j < n_plp[i]; ++j) { const bam_pileup1_t *p = plp[i] + j; // DON'T modfity plp[][] unless you really know if (p->is_del || p->is_refskip) ++m; // having dels or refskips at tid:pos else if (bam1_qual(p->b)[p->qpos] < baseQ) ++m; // low base quality } printf("\t%d", n_plp[i] - m); // this the depth to output } putchar('\n'); } free(n_plp); free(plp); bam_mplp_destroy(mplp); bam_header_destroy(h); for (i = 0; i < n; ++i) { bam_close(data[i]->fp); if (data[i]->iter) bam_iter_destroy(data[i]->iter); free(data[i]); } free(data); free(reg); if (bed) bed_destroy(bed); if ( file_list ) { for (i=0; i