#include #include #include #include #include "sam.h" typedef struct __linkbuf_t { bam1_t b; uint32_t beg, end; struct __linkbuf_t *next; } lbnode_t; /* --- BEGIN: Memory pool */ typedef struct { int cnt, n, max; lbnode_t **buf; } mempool_t; static mempool_t *mp_init() { mempool_t *mp; mp = (mempool_t*)calloc(1, sizeof(mempool_t)); return mp; } static void mp_destroy(mempool_t *mp) { int k; for (k = 0; k < mp->n; ++k) { free(mp->buf[k]->b.data); free(mp->buf[k]); } free(mp->buf); free(mp); } static inline lbnode_t *mp_alloc(mempool_t *mp) { ++mp->cnt; if (mp->n == 0) return (lbnode_t*)calloc(1, sizeof(lbnode_t)); else return mp->buf[--mp->n]; } static inline void mp_free(mempool_t *mp, lbnode_t *p) { --mp->cnt; p->next = 0; // clear lbnode_t::next here if (mp->n == mp->max) { mp->max = mp->max? mp->max<<1 : 256; mp->buf = (lbnode_t**)realloc(mp->buf, sizeof(lbnode_t*) * mp->max); } mp->buf[mp->n++] = p; } /* --- END: Memory pool */ /* --- BEGIN: Auxiliary functions */ static inline int resolve_cigar(bam_pileup1_t *p, uint32_t pos) { unsigned k; bam1_t *b = p->b; bam1_core_t *c = &b->core; uint32_t x = c->pos, y = 0; int ret = 1, is_restart = 1; if (c->flag&BAM_FUNMAP) return 0; // unmapped read assert(x <= pos); // otherwise a bug p->qpos = -1; p->indel = 0; p->is_del = p->is_head = p->is_tail = 0; for (k = 0; k < c->n_cigar; ++k) { int op = bam1_cigar(b)[k] & BAM_CIGAR_MASK; // operation int l = bam1_cigar(b)[k] >> BAM_CIGAR_SHIFT; // length if (op == BAM_CMATCH) { // NOTE: this assumes the first and the last operation MUST BE a match or a clip if (x + l > pos) { // overlap with pos p->indel = p->is_del = 0; p->qpos = y + (pos - x); if (x == pos && is_restart) p->is_head = 1; if (x + l - 1 == pos) { // come to the end of a match int has_next_match = 0; unsigned i; for (i = k + 1; i < c->n_cigar; ++i) { uint32_t cigar = bam1_cigar(b)[i]; int opi = cigar&BAM_CIGAR_MASK; if (opi == BAM_CMATCH) { has_next_match = 1; break; } else if (opi == BAM_CSOFT_CLIP || opi == BAM_CREF_SKIP || opi == BAM_CHARD_CLIP) break; } if (!has_next_match) p->is_tail = 1; if (k < c->n_cigar - 1 && has_next_match) { // there are additional operation(s) uint32_t cigar = bam1_cigar(b)[k+1]; // next CIGAR int op_next = cigar&BAM_CIGAR_MASK; // next CIGAR operation if (op_next == BAM_CDEL) p->indel = -(int32_t)(cigar>>BAM_CIGAR_SHIFT); // del else if (op_next == BAM_CINS) p->indel = cigar>>BAM_CIGAR_SHIFT; // ins else if (op_next == BAM_CPAD && k + 2 < c->n_cigar) { // no working for adjacent padding cigar = bam1_cigar(b)[k+2]; op_next = cigar&BAM_CIGAR_MASK; if (op_next == BAM_CDEL) p->indel = -(int32_t)(cigar>>BAM_CIGAR_SHIFT); // del else if (op_next == BAM_CINS) p->indel = cigar>>BAM_CIGAR_SHIFT; // ins } } } } x += l; y += l; } else if (op == BAM_CDEL) { // then set ->is_del if (x + l > pos) { p->indel = 0; p->is_del = 1; p->qpos = y + (pos - x); } x += l; } else if (op == BAM_CREF_SKIP) x += l; else if (op == BAM_CINS || op == BAM_CSOFT_CLIP) y += l; if (is_restart) is_restart ^= (op == BAM_CMATCH); else is_restart ^= (op == BAM_CREF_SKIP || op == BAM_CSOFT_CLIP || op == BAM_CHARD_CLIP); if (x > pos) { if (op == BAM_CREF_SKIP) ret = 0; // then do not put it into pileup at all break; } } assert(x > pos); // otherwise a bug return ret; } /* --- END: Auxiliary functions */ /******************* * pileup iterator * *******************/ struct __bam_plp_t { mempool_t *mp; lbnode_t *head, *tail, *dummy; int32_t tid, pos, max_tid, max_pos; int is_eof, flag_mask, max_plp, error; bam_pileup1_t *plp; // for the "auto" interface only bam1_t *b; bam_plp_auto_f func; void *data; }; bam_plp_t bam_plp_init(bam_plp_auto_f func, void *data) { bam_plp_t iter; iter = calloc(1, sizeof(struct __bam_plp_t)); iter->mp = mp_init(); iter->head = iter->tail = mp_alloc(iter->mp); iter->dummy = mp_alloc(iter->mp); iter->max_tid = iter->max_pos = -1; iter->flag_mask = BAM_DEF_MASK; if (func) { iter->func = func; iter->data = data; iter->b = bam_init1(); } return iter; } void bam_plp_destroy(bam_plp_t iter) { mp_free(iter->mp, iter->dummy); mp_free(iter->mp, iter->head); if (iter->mp->cnt != 0) fprintf(stderr, "[bam_plp_destroy] memory leak: %d. Continue anyway.\n", iter->mp->cnt); mp_destroy(iter->mp); if (iter->b) bam_destroy1(iter->b); free(iter->plp); free(iter); } const bam_pileup1_t *bam_plp_next(bam_plp_t iter, int *_tid, int *_pos, int *_n_plp) { if (iter->error) { *_n_plp = -1; return 0; } *_n_plp = 0; if (iter->is_eof && iter->head->next == 0) return 0; while (iter->is_eof || iter->max_tid > iter->tid || (iter->max_tid == iter->tid && iter->max_pos > iter->pos)) { int n_plp = 0; lbnode_t *p, *q; // write iter->plp at iter->pos iter->dummy->next = iter->head; for (p = iter->head, q = iter->dummy; p->next; q = p, p = p->next) { if (p->b.core.tid < iter->tid || (p->b.core.tid == iter->tid && p->end <= iter->pos)) { // then remove q->next = p->next; mp_free(iter->mp, p); p = q; } else if (p->b.core.tid == iter->tid && p->beg <= iter->pos) { // here: p->end > pos; then add to pileup if (n_plp == iter->max_plp) { // then double the capacity iter->max_plp = iter->max_plp? iter->max_plp<<1 : 256; iter->plp = (bam_pileup1_t*)realloc(iter->plp, sizeof(bam_pileup1_t) * iter->max_plp); } iter->plp[n_plp].b = &p->b; if (resolve_cigar(iter->plp + n_plp, iter->pos)) ++n_plp; // skip the read if we are looking at ref-skip } } iter->head = iter->dummy->next; // dummy->next may be changed *_n_plp = n_plp; *_tid = iter->tid; *_pos = iter->pos; // update iter->tid and iter->pos if (iter->head->next) { if (iter->tid > iter->head->b.core.tid) { fprintf(stderr, "[%s] unsorted input. Pileup aborts.\n", __func__); iter->error = 1; *_n_plp = -1; return 0; } } if (iter->tid < iter->head->b.core.tid) { // come to a new reference sequence iter->tid = iter->head->b.core.tid; iter->pos = iter->head->beg; // jump to the next reference } else if (iter->pos < iter->head->beg) { // here: tid == head->b.core.tid iter->pos = iter->head->beg; // jump to the next position } else ++iter->pos; // scan contiguously // return if (n_plp) return iter->plp; if (iter->is_eof && iter->head->next == 0) break; } return 0; } int bam_plp_push(bam_plp_t iter, const bam1_t *b) { if (iter->error) return -1; if (b) { if (b->core.tid < 0) return 0; if (b->core.flag & iter->flag_mask) return 0; bam_copy1(&iter->tail->b, b); iter->tail->beg = b->core.pos; iter->tail->end = bam_calend(&b->core, bam1_cigar(b)); if (b->core.tid < iter->max_tid) { fprintf(stderr, "[bam_pileup_core] the input is not sorted (chromosomes out of order)\n"); iter->error = 1; return -1; } if ((b->core.tid == iter->max_tid) && (iter->tail->beg < iter->max_pos)) { fprintf(stderr, "[bam_pileup_core] the input is not sorted (reads out of order)\n"); iter->error = 1; return -1; } iter->max_tid = b->core.tid; iter->max_pos = iter->tail->beg; if (iter->tail->end > iter->pos || iter->tail->b.core.tid > iter->tid) { iter->tail->next = mp_alloc(iter->mp); iter->tail = iter->tail->next; } } else iter->is_eof = 1; return 0; } const bam_pileup1_t *bam_plp_auto(bam_plp_t iter, int *_tid, int *_pos, int *_n_plp) { const bam_pileup1_t *plp; if (iter->func == 0 || iter->error) { *_n_plp = -1; return 0; } if ((plp = bam_plp_next(iter, _tid, _pos, _n_plp)) != 0) return plp; else { *_n_plp = 0; if (iter->is_eof) return 0; while (iter->func(iter->data, iter->b) >= 0) { if (bam_plp_push(iter, iter->b) < 0) { *_n_plp = -1; return 0; } if ((plp = bam_plp_next(iter, _tid, _pos, _n_plp)) != 0) return plp; } bam_plp_push(iter, 0); if ((plp = bam_plp_next(iter, _tid, _pos, _n_plp)) != 0) return plp; return 0; } } void bam_plp_reset(bam_plp_t iter) { lbnode_t *p, *q; iter->max_tid = iter->max_pos = -1; iter->tid = iter->pos = 0; iter->is_eof = 0; for (p = iter->head; p->next;) { q = p->next; mp_free(iter->mp, p); p = q; } iter->head = iter->tail; } void bam_plp_set_mask(bam_plp_t iter, int mask) { iter->flag_mask = mask < 0? BAM_DEF_MASK : (BAM_FUNMAP | mask); } /***************** * callback APIs * *****************/ int bam_pileup_file(bamFile fp, int mask, bam_pileup_f func, void *func_data) { bam_plbuf_t *buf; int ret; bam1_t *b; b = bam_init1(); buf = bam_plbuf_init(func, func_data); bam_plbuf_set_mask(buf, mask); while ((ret = bam_read1(fp, b)) >= 0) bam_plbuf_push(b, buf); bam_plbuf_push(0, buf); bam_plbuf_destroy(buf); bam_destroy1(b); return 0; } void bam_plbuf_set_mask(bam_plbuf_t *buf, int mask) { bam_plp_set_mask(buf->iter, mask); } void bam_plbuf_reset(bam_plbuf_t *buf) { bam_plp_reset(buf->iter); } bam_plbuf_t *bam_plbuf_init(bam_pileup_f func, void *data) { bam_plbuf_t *buf; buf = calloc(1, sizeof(bam_plbuf_t)); buf->iter = bam_plp_init(0, 0); buf->func = func; buf->data = data; return buf; } void bam_plbuf_destroy(bam_plbuf_t *buf) { bam_plp_destroy(buf->iter); free(buf); } int bam_plbuf_push(const bam1_t *b, bam_plbuf_t *buf) { int ret, n_plp, tid, pos; const bam_pileup1_t *plp; ret = bam_plp_push(buf->iter, b); if (ret < 0) return ret; while ((plp = bam_plp_next(buf->iter, &tid, &pos, &n_plp)) != 0) buf->func(tid, pos, n_plp, plp, buf->data); return 0; } /*********** * mpileup * ***********/ struct __bam_mplp_t { int n; uint64_t min, *pos; bam_plp_t *iter; int *n_plp; const bam_pileup1_t **plp; }; bam_mplp_t bam_mplp_init(int n, bam_plp_auto_f func, void **data) { int i; bam_mplp_t iter; iter = calloc(1, sizeof(struct __bam_mplp_t)); iter->pos = calloc(n, 8); iter->n_plp = calloc(n, sizeof(int)); iter->plp = calloc(n, sizeof(void*)); iter->iter = calloc(n, sizeof(void*)); iter->n = n; iter->min = (uint64_t)-1; for (i = 0; i < n; ++i) { iter->iter[i] = bam_plp_init(func, data[i]); iter->pos[i] = iter->min; } return iter; } void bam_mplp_destroy(bam_mplp_t iter) { int i; for (i = 0; i < iter->n; ++i) bam_plp_destroy(iter->iter[i]); free(iter->iter); free(iter->pos); free(iter->n_plp); free(iter->plp); free(iter); } int bam_mplp_auto(bam_mplp_t iter, int *_tid, int *_pos, int *n_plp, const bam_pileup1_t **plp) { int i, ret = 0; uint64_t new_min = (uint64_t)-1; for (i = 0; i < iter->n; ++i) { if (iter->pos[i] == iter->min) { int tid, pos; iter->plp[i] = bam_plp_auto(iter->iter[i], &tid, &pos, &iter->n_plp[i]); iter->pos[i] = (uint64_t)tid<<32 | pos; } if (iter->plp[i] && iter->pos[i] < new_min) new_min = iter->pos[i]; } iter->min = new_min; if (new_min == (uint64_t)-1) return 0; *_tid = new_min>>32; *_pos = (uint32_t)new_min; for (i = 0; i < iter->n; ++i) { if (iter->pos[i] == iter->min) { n_plp[i] = iter->n_plp[i], plp[i] = iter->plp[i]; ++ret; } else n_plp[i] = 0, plp[i] = 0; } return ret; }