#include #include #include #include #include #include "sam.h" typedef bam1_t *bam1_p; #include "khash.h" KHASH_SET_INIT_STR(name) KHASH_MAP_INIT_INT64(pos, bam1_p) #define BUFFER_SIZE 0x40000 typedef struct { uint64_t n_checked, n_removed; khash_t(pos) *best_hash; } lib_aux_t; KHASH_MAP_INIT_STR(lib, lib_aux_t) typedef struct { int n, max; bam1_t **a; } tmp_stack_t; static inline void stack_insert(tmp_stack_t *stack, bam1_t *b) { if (stack->n == stack->max) { stack->max = stack->max? stack->max<<1 : 0x10000; stack->a = (bam1_t**)realloc(stack->a, sizeof(bam1_t*) * stack->max); } stack->a[stack->n++] = b; } static inline void dump_best(tmp_stack_t *stack, samfile_t *out) { int i; for (i = 0; i != stack->n; ++i) { samwrite(out, stack->a[i]); bam_destroy1(stack->a[i]); } stack->n = 0; } static void clear_del_set(khash_t(name) *del_set) { khint_t k; for (k = kh_begin(del_set); k < kh_end(del_set); ++k) if (kh_exist(del_set, k)) free((char*)kh_key(del_set, k)); kh_clear(name, del_set); } static lib_aux_t *get_aux(khash_t(lib) *aux, const char *lib) { khint_t k = kh_get(lib, aux, lib); if (k == kh_end(aux)) { int ret; char *p = strdup(lib); lib_aux_t *q; k = kh_put(lib, aux, p, &ret); q = &kh_val(aux, k); q->n_checked = q->n_removed = 0; q->best_hash = kh_init(pos); return q; } else return &kh_val(aux, k); } static void clear_best(khash_t(lib) *aux, int max) { khint_t k; for (k = kh_begin(aux); k != kh_end(aux); ++k) { if (kh_exist(aux, k)) { lib_aux_t *q = &kh_val(aux, k); if (kh_size(q->best_hash) >= max) kh_clear(pos, q->best_hash); } } } static inline int sum_qual(const bam1_t *b) { int i, q; uint8_t *qual = bam1_qual(b); for (i = q = 0; i < b->core.l_qseq; ++i) q += qual[i]; return q; } void bam_rmdup_core(samfile_t *in, samfile_t *out) { bam1_t *b; int last_tid = -1, last_pos = -1; tmp_stack_t stack; khint_t k; khash_t(lib) *aux; khash_t(name) *del_set; aux = kh_init(lib); del_set = kh_init(name); b = bam_init1(); memset(&stack, 0, sizeof(tmp_stack_t)); kh_resize(name, del_set, 4 * BUFFER_SIZE); while (samread(in, b) >= 0) { bam1_core_t *c = &b->core; if (c->tid != last_tid || last_pos != c->pos) { dump_best(&stack, out); // write the result clear_best(aux, BUFFER_SIZE); if (c->tid != last_tid) { clear_best(aux, 0); if (kh_size(del_set)) { // check fprintf(stderr, "[bam_rmdup_core] %llu unmatched pairs\n", (long long)kh_size(del_set)); clear_del_set(del_set); } if ((int)c->tid == -1) { // append unmapped reads samwrite(out, b); while (samread(in, b) >= 0) samwrite(out, b); break; } last_tid = c->tid; fprintf(stderr, "[bam_rmdup_core] processing reference %s...\n", in->header->target_name[c->tid]); } } if (!(c->flag&BAM_FPAIRED) || (c->flag&(BAM_FUNMAP|BAM_FMUNMAP)) || (c->mtid >= 0 && c->tid != c->mtid)) { samwrite(out, b); } else if (c->isize > 0) { // paired, head uint64_t key = (uint64_t)c->pos<<32 | c->isize; const char *lib; lib_aux_t *q; int ret; lib = bam_get_library(in->header, b); q = lib? get_aux(aux, lib) : get_aux(aux, "\t"); ++q->n_checked; k = kh_put(pos, q->best_hash, key, &ret); if (ret == 0) { // found in best_hash bam1_t *p = kh_val(q->best_hash, k); ++q->n_removed; if (sum_qual(p) < sum_qual(b)) { // the current alignment is better; this can be accelerated in principle kh_put(name, del_set, strdup(bam1_qname(p)), &ret); // p will be removed bam_copy1(p, b); // replaced as b } else kh_put(name, del_set, strdup(bam1_qname(b)), &ret); // b will be removed if (ret == 0) fprintf(stderr, "[bam_rmdup_core] inconsistent BAM file for pair '%s'. Continue anyway.\n", bam1_qname(b)); } else { // not found in best_hash kh_val(q->best_hash, k) = bam_dup1(b); stack_insert(&stack, kh_val(q->best_hash, k)); } } else { // paired, tail k = kh_get(name, del_set, bam1_qname(b)); if (k != kh_end(del_set)) { free((char*)kh_key(del_set, k)); kh_del(name, del_set, k); } else samwrite(out, b); } last_pos = c->pos; } for (k = kh_begin(aux); k != kh_end(aux); ++k) { if (kh_exist(aux, k)) { lib_aux_t *q = &kh_val(aux, k); dump_best(&stack, out); fprintf(stderr, "[bam_rmdup_core] %lld / %lld = %.4lf in library '%s'\n", (long long)q->n_removed, (long long)q->n_checked, (double)q->n_removed/q->n_checked, kh_key(aux, k)); kh_destroy(pos, q->best_hash); free((char*)kh_key(aux, k)); } } kh_destroy(lib, aux); clear_del_set(del_set); kh_destroy(name, del_set); free(stack.a); bam_destroy1(b); } void bam_rmdupse_core(samfile_t *in, samfile_t *out, int force_se); int bam_rmdup(int argc, char *argv[]) { int c, is_se = 0, force_se = 0; samfile_t *in, *out; while ((c = getopt(argc, argv, "sS")) >= 0) { switch (c) { case 's': is_se = 1; break; case 'S': force_se = is_se = 1; break; } } if (optind + 2 > argc) { fprintf(stderr, "\n"); fprintf(stderr, "Usage: samtools rmdup [-sS] \n\n"); fprintf(stderr, "Option: -s rmdup for SE reads\n"); fprintf(stderr, " -S treat PE reads as SE in rmdup (force -s)\n\n"); return 1; } in = samopen(argv[optind], "rb", 0); out = samopen(argv[optind+1], "wb", in->header); if (in == 0 || out == 0) { fprintf(stderr, "[bam_rmdup] fail to read/write input files\n"); return 1; } if (is_se) bam_rmdupse_core(in, out, force_se); else bam_rmdup_core(in, out); samclose(in); samclose(out); return 0; }