#include #include #include #include #include "kthread.h" #include "kvec.h" #include "kalloc.h" #include "sdust.h" #include "mmpriv.h" #include "bseq.h" #include "khash.h" struct mm_tbuf_s { void *km; int rep_len, frag_gap; }; mm_tbuf_t *mm_tbuf_init(void) { mm_tbuf_t *b; b = (mm_tbuf_t*)calloc(1, sizeof(mm_tbuf_t)); if (!(mm_dbg_flag & 1)) b->km = km_init(); return b; } void mm_tbuf_destroy(mm_tbuf_t *b) { if (b == 0) return; km_destroy(b->km); free(b); } void *mm_tbuf_get_km(mm_tbuf_t *b) { return b->km; } static int mm_dust_minier(void *km, int n, mm128_t *a, int l_seq, const char *seq, int sdust_thres) { int n_dreg, j, k, u = 0; const uint64_t *dreg; sdust_buf_t *sdb; if (sdust_thres <= 0) return n; sdb = sdust_buf_init(km); dreg = sdust_core((const uint8_t*)seq, l_seq, sdust_thres, 64, &n_dreg, sdb); for (j = k = 0; j < n; ++j) { // squeeze out minimizers that significantly overlap with LCRs int32_t qpos = (uint32_t)a[j].y>>1, span = a[j].x&0xff; int32_t s = qpos - (span - 1), e = s + span; while (u < n_dreg && (int32_t)dreg[u] <= s) ++u; if (u < n_dreg && (int32_t)(dreg[u]>>32) < e) { int v, l = 0; for (v = u; v < n_dreg && (int32_t)(dreg[v]>>32) < e; ++v) { // iterate over LCRs overlapping this minimizer int ss = s > (int32_t)(dreg[v]>>32)? s : dreg[v]>>32; int ee = e < (int32_t)dreg[v]? e : (uint32_t)dreg[v]; l += ee - ss; } if (l <= span>>1) a[k++] = a[j]; // keep the minimizer if less than half of it falls in masked region } else a[k++] = a[j]; } sdust_buf_destroy(sdb); return k; // the new size } static void collect_minimizers(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int n_segs, const int *qlens, const char **seqs, mm128_v *mv) { int i, n, sum = 0; mv->n = 0; for (i = n = 0; i < n_segs; ++i) { size_t j; mm_sketch(km, seqs[i], qlens[i], mi->w, mi->k, i, mi->flag&MM_I_HPC, mv); for (j = n; j < mv->n; ++j) mv->a[j].y += sum << 1; if (opt->sdust_thres > 0) // mask low-complexity minimizers mv->n = n + mm_dust_minier(km, mv->n - n, mv->a + n, qlens[i], seqs[i], opt->sdust_thres); sum += qlens[i], n = mv->n; } } #include "ksort.h" #define heap_lt(a, b) ((a).x > (b).x) KSORT_INIT(heap, mm128_t, heap_lt) static inline int skip_seed(int flag, uint64_t r, const mm_seed_t *q, const char *qname, int qlen, const mm_idx_t *mi, int *is_self) { *is_self = 0; if (qname && (flag & (MM_F_NO_DIAG|MM_F_NO_DUAL))) { const mm_idx_seq_t *s = &mi->seq[r>>32]; int cmp; cmp = strcmp(qname, s->name); if ((flag&MM_F_NO_DIAG) && cmp == 0 && (int)s->len == qlen) { if ((uint32_t)r>>1 == (q->q_pos>>1)) return 1; // avoid the diagnonal anchors if ((r&1) == (q->q_pos&1)) *is_self = 1; // this flag is used to avoid spurious extension on self chain } if ((flag&MM_F_NO_DUAL) && cmp > 0) // all-vs-all mode: map once return 1; } if (flag & (MM_F_FOR_ONLY|MM_F_REV_ONLY)) { if ((r&1) == (q->q_pos&1)) { // forward strand if (flag & MM_F_REV_ONLY) return 1; } else { if (flag & MM_F_FOR_ONLY) return 1; } } return 0; } static mm128_t *collect_seed_hits_heap(void *km, const mm_mapopt_t *opt, int max_occ, const mm_idx_t *mi, const char *qname, const mm128_v *mv, int qlen, int64_t *n_a, int *rep_len, int *n_mini_pos, uint64_t **mini_pos) { int i, n_m, heap_size = 0; int64_t j, n_for = 0, n_rev = 0; mm_seed_t *m; mm128_t *a, *heap; m = mm_collect_matches(km, &n_m, qlen, max_occ, opt->max_max_occ, opt->occ_dist, mi, mv, n_a, rep_len, n_mini_pos, mini_pos); heap = (mm128_t*)kmalloc(km, n_m * sizeof(mm128_t)); a = (mm128_t*)kmalloc(km, *n_a * sizeof(mm128_t)); for (i = 0, heap_size = 0; i < n_m; ++i) { if (m[i].n > 0) { heap[heap_size].x = m[i].cr[0]; heap[heap_size].y = (uint64_t)i<<32; ++heap_size; } } ks_heapmake_heap(heap_size, heap); while (heap_size > 0) { mm_seed_t *q = &m[heap->y>>32]; mm128_t *p; uint64_t r = heap->x; int32_t is_self, rpos = (uint32_t)r >> 1; if (!skip_seed(opt->flag, r, q, qname, qlen, mi, &is_self)) { if ((r&1) == (q->q_pos&1)) { // forward strand p = &a[n_for++]; p->x = (r&0xffffffff00000000ULL) | rpos; p->y = (uint64_t)q->q_span << 32 | q->q_pos >> 1; } else { // reverse strand p = &a[(*n_a) - (++n_rev)]; p->x = 1ULL<<63 | (r&0xffffffff00000000ULL) | rpos; p->y = (uint64_t)q->q_span << 32 | (qlen - ((q->q_pos>>1) + 1 - q->q_span) - 1); } p->y |= (uint64_t)q->seg_id << MM_SEED_SEG_SHIFT; if (q->is_tandem) p->y |= MM_SEED_TANDEM; if (is_self) p->y |= MM_SEED_SELF; } // update the heap if ((uint32_t)heap->y < q->n - 1) { ++heap[0].y; heap[0].x = m[heap[0].y>>32].cr[(uint32_t)heap[0].y]; } else { heap[0] = heap[heap_size - 1]; --heap_size; } ks_heapdown_heap(0, heap_size, heap); } kfree(km, m); kfree(km, heap); // reverse anchors on the reverse strand, as they are in the descending order for (j = 0; j < n_rev>>1; ++j) { mm128_t t = a[(*n_a) - 1 - j]; a[(*n_a) - 1 - j] = a[(*n_a) - (n_rev - j)]; a[(*n_a) - (n_rev - j)] = t; } if (*n_a > n_for + n_rev) { memmove(a + n_for, a + (*n_a) - n_rev, n_rev * sizeof(mm128_t)); *n_a = n_for + n_rev; } return a; } static mm128_t *collect_seed_hits(void *km, const mm_mapopt_t *opt, int max_occ, const mm_idx_t *mi, const char *qname, const mm128_v *mv, int qlen, int64_t *n_a, int *rep_len, int *n_mini_pos, uint64_t **mini_pos) { int i, n_m; mm_seed_t *m; mm128_t *a; m = mm_collect_matches(km, &n_m, qlen, max_occ, opt->max_max_occ, opt->occ_dist, mi, mv, n_a, rep_len, n_mini_pos, mini_pos); a = (mm128_t*)kmalloc(km, *n_a * sizeof(mm128_t)); for (i = 0, *n_a = 0; i < n_m; ++i) { mm_seed_t *q = &m[i]; const uint64_t *r = q->cr; uint32_t k; for (k = 0; k < q->n; ++k) { int32_t is_self, rpos = (uint32_t)r[k] >> 1; mm128_t *p; if (skip_seed(opt->flag, r[k], q, qname, qlen, mi, &is_self)) continue; p = &a[(*n_a)++]; if ((r[k]&1) == (q->q_pos&1)) { // forward strand p->x = (r[k]&0xffffffff00000000ULL) | rpos; p->y = (uint64_t)q->q_span << 32 | q->q_pos >> 1; } else if (!(opt->flag & MM_F_QSTRAND)) { // reverse strand and not in the query-strand mode p->x = 1ULL<<63 | (r[k]&0xffffffff00000000ULL) | rpos; p->y = (uint64_t)q->q_span << 32 | (qlen - ((q->q_pos>>1) + 1 - q->q_span) - 1); } else { // reverse strand; query-strand int32_t len = mi->seq[r[k]>>32].len; p->x = 1ULL<<63 | (r[k]&0xffffffff00000000ULL) | (len - (rpos + 1 - q->q_span) - 1); // coordinate only accurate for non-HPC seeds p->y = (uint64_t)q->q_span << 32 | q->q_pos >> 1; } p->y |= (uint64_t)q->seg_id << MM_SEED_SEG_SHIFT; if (q->is_tandem) p->y |= MM_SEED_TANDEM; if (is_self) p->y |= MM_SEED_SELF; } } kfree(km, m); radix_sort_128x(a, a + (*n_a)); return a; } static void chain_post(const mm_mapopt_t *opt, int max_chain_gap_ref, const mm_idx_t *mi, void *km, int qlen, int n_segs, const int *qlens, int *n_regs, mm_reg1_t *regs, mm128_t *a) { if (!(opt->flag & MM_F_ALL_CHAINS)) { // don't choose primary mapping(s) mm_set_parent(km, opt->mask_level, opt->mask_len, *n_regs, regs, opt->a * 2 + opt->b, opt->flag&MM_F_HARD_MLEVEL, opt->alt_drop); if (n_segs <= 1) mm_select_sub(km, opt->pri_ratio, mi->k*2, opt->best_n, 1, opt->max_gap * 0.8, n_regs, regs); else mm_select_sub_multi(km, opt->pri_ratio, 0.2f, 0.7f, max_chain_gap_ref, mi->k*2, opt->best_n, n_segs, qlens, n_regs, regs); } } static mm_reg1_t *align_regs(const mm_mapopt_t *opt, const mm_idx_t *mi, void *km, int qlen, const char *seq, int *n_regs, mm_reg1_t *regs, mm128_t *a) { if (!(opt->flag & MM_F_CIGAR)) return regs; regs = mm_align_skeleton(km, opt, mi, qlen, seq, n_regs, regs, a); // this calls mm_filter_regs() if (!(opt->flag & MM_F_ALL_CHAINS)) { // don't choose primary mapping(s) mm_set_parent(km, opt->mask_level, opt->mask_len, *n_regs, regs, opt->a * 2 + opt->b, opt->flag&MM_F_HARD_MLEVEL, opt->alt_drop); mm_select_sub(km, opt->pri_ratio, mi->k*2, opt->best_n, 0, opt->max_gap * 0.8, n_regs, regs); mm_set_sam_pri(*n_regs, regs); } return regs; } void mm_map_frag(const mm_idx_t *mi, int n_segs, const int *qlens, const char **seqs, int *n_regs, mm_reg1_t **regs, mm_tbuf_t *b, const mm_mapopt_t *opt, const char *qname) { int i, j, rep_len, qlen_sum, n_regs0, n_mini_pos; int max_chain_gap_qry, max_chain_gap_ref, is_splice = !!(opt->flag & MM_F_SPLICE), is_sr = !!(opt->flag & MM_F_SR); uint32_t hash; int64_t n_a; uint64_t *u, *mini_pos; mm128_t *a; mm128_v mv = {0,0,0}; mm_reg1_t *regs0; km_stat_t kmst; float chn_pen_gap, chn_pen_skip; for (i = 0, qlen_sum = 0; i < n_segs; ++i) qlen_sum += qlens[i], n_regs[i] = 0, regs[i] = 0; if (qlen_sum == 0 || n_segs <= 0 || n_segs > MM_MAX_SEG) return; if (opt->max_qlen > 0 && qlen_sum > opt->max_qlen) return; hash = qname && !(opt->flag & MM_F_NO_HASH_NAME)? __ac_X31_hash_string(qname) : 0; hash ^= __ac_Wang_hash(qlen_sum) + __ac_Wang_hash(opt->seed); hash = __ac_Wang_hash(hash); collect_minimizers(b->km, opt, mi, n_segs, qlens, seqs, &mv); if (opt->q_occ_frac > 0.0f) mm_seed_mz_flt(b->km, &mv, opt->mid_occ, opt->q_occ_frac); if (opt->flag & MM_F_HEAP_SORT) a = collect_seed_hits_heap(b->km, opt, opt->mid_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos); else a = collect_seed_hits(b->km, opt, opt->mid_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos); if (mm_dbg_flag & MM_DBG_PRINT_SEED) { fprintf(stderr, "RS\t%d\n", rep_len); for (i = 0; i < n_a; ++i) fprintf(stderr, "SD\t%s\t%d\t%c\t%d\t%d\t%d\n", mi->seq[a[i].x<<1>>33].name, (int32_t)a[i].x, "+-"[a[i].x>>63], (int32_t)a[i].y, (int32_t)(a[i].y>>32&0xff), i == 0? 0 : ((int32_t)a[i].y - (int32_t)a[i-1].y) - ((int32_t)a[i].x - (int32_t)a[i-1].x)); } // set max chaining gap on the query and the reference sequence if (is_sr) max_chain_gap_qry = qlen_sum > opt->max_gap? qlen_sum : opt->max_gap; else max_chain_gap_qry = opt->max_gap; if (opt->max_gap_ref > 0) { max_chain_gap_ref = opt->max_gap_ref; // always honor mm_mapopt_t::max_gap_ref if set } else if (opt->max_frag_len > 0) { max_chain_gap_ref = opt->max_frag_len - qlen_sum; if (max_chain_gap_ref < opt->max_gap) max_chain_gap_ref = opt->max_gap; } else max_chain_gap_ref = opt->max_gap; chn_pen_gap = opt->chain_gap_scale * 0.01 * mi->k; chn_pen_skip = opt->chain_skip_scale * 0.01 * mi->k; if (opt->flag & MM_F_RMQ) { a = mg_lchain_rmq(opt->max_gap, opt->rmq_inner_dist, opt->bw, opt->max_chain_skip, opt->rmq_size_cap, opt->min_cnt, opt->min_chain_score, chn_pen_gap, chn_pen_skip, n_a, a, &n_regs0, &u, b->km); } else { a = mg_lchain_dp(max_chain_gap_ref, max_chain_gap_qry, opt->bw, opt->max_chain_skip, opt->max_chain_iter, opt->min_cnt, opt->min_chain_score, chn_pen_gap, chn_pen_skip, is_splice, n_segs, n_a, a, &n_regs0, &u, b->km); } if (opt->bw_long > opt->bw && (opt->flag & (MM_F_SPLICE|MM_F_SR|MM_F_NO_LJOIN)) == 0 && n_segs == 1 && n_regs0 > 1) { // re-chain/long-join for long sequences int32_t st = (int32_t)a[0].y, en = (int32_t)a[(int32_t)u[0] - 1].y; if (qlen_sum - (en - st) > opt->rmq_rescue_size || en - st > qlen_sum * opt->rmq_rescue_ratio) { int32_t i; for (i = 0, n_a = 0; i < n_regs0; ++i) n_a += (int32_t)u[i]; kfree(b->km, u); radix_sort_128x(a, a + n_a); a = mg_lchain_rmq(opt->max_gap, opt->rmq_inner_dist, opt->bw_long, opt->max_chain_skip, opt->rmq_size_cap, opt->min_cnt, opt->min_chain_score, chn_pen_gap, chn_pen_skip, n_a, a, &n_regs0, &u, b->km); } } else if (opt->max_occ > opt->mid_occ && rep_len > 0 && !(opt->flag & MM_F_RMQ)) { // re-chain, mostly for short reads int rechain = 0; if (n_regs0 > 0) { // test if the best chain has all the segments int n_chained_segs = 1, max = 0, max_i = -1, max_off = -1, off = 0; for (i = 0; i < n_regs0; ++i) { // find the best chain if (max < (int)(u[i]>>32)) max = u[i]>>32, max_i = i, max_off = off; off += (uint32_t)u[i]; } for (i = 1; i < (int32_t)u[max_i]; ++i) // count the number of segments in the best chain if ((a[max_off+i].y&MM_SEED_SEG_MASK) != (a[max_off+i-1].y&MM_SEED_SEG_MASK)) ++n_chained_segs; if (n_chained_segs < n_segs) rechain = 1; } else rechain = 1; if (rechain) { // redo chaining with a higher max_occ threshold kfree(b->km, a); kfree(b->km, u); kfree(b->km, mini_pos); if (opt->flag & MM_F_HEAP_SORT) a = collect_seed_hits_heap(b->km, opt, opt->max_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos); else a = collect_seed_hits(b->km, opt, opt->max_occ, mi, qname, &mv, qlen_sum, &n_a, &rep_len, &n_mini_pos, &mini_pos); a = mg_lchain_dp(max_chain_gap_ref, max_chain_gap_qry, opt->bw, opt->max_chain_skip, opt->max_chain_iter, opt->min_cnt, opt->min_chain_score, chn_pen_gap, chn_pen_skip, is_splice, n_segs, n_a, a, &n_regs0, &u, b->km); } } b->frag_gap = max_chain_gap_ref; b->rep_len = rep_len; regs0 = mm_gen_regs(b->km, hash, qlen_sum, n_regs0, u, a, !!(opt->flag&MM_F_QSTRAND)); if (mi->n_alt) { mm_mark_alt(mi, n_regs0, regs0); mm_hit_sort(b->km, &n_regs0, regs0, opt->alt_drop); // this step can be merged into mm_gen_regs(); will do if this shows up in profile } if (mm_dbg_flag & (MM_DBG_PRINT_SEED|MM_DBG_PRINT_CHAIN)) for (j = 0; j < n_regs0; ++j) for (i = regs0[j].as; i < regs0[j].as + regs0[j].cnt; ++i) fprintf(stderr, "CN\t%d\t%s\t%d\t%c\t%d\t%d\t%d\n", j, mi->seq[a[i].x<<1>>33].name, (int32_t)a[i].x, "+-"[a[i].x>>63], (int32_t)a[i].y, (int32_t)(a[i].y>>32&0xff), i == regs0[j].as? 0 : ((int32_t)a[i].y - (int32_t)a[i-1].y) - ((int32_t)a[i].x - (int32_t)a[i-1].x)); chain_post(opt, max_chain_gap_ref, mi, b->km, qlen_sum, n_segs, qlens, &n_regs0, regs0, a); if (!is_sr && !(opt->flag&MM_F_QSTRAND)) { mm_est_err(mi, qlen_sum, n_regs0, regs0, a, n_mini_pos, mini_pos); n_regs0 = mm_filter_strand_retained(n_regs0, regs0); } if (n_segs == 1) { // uni-segment regs0 = align_regs(opt, mi, b->km, qlens[0], seqs[0], &n_regs0, regs0, a); regs0 = (mm_reg1_t*)realloc(regs0, sizeof(*regs0) * n_regs0); mm_set_mapq(b->km, n_regs0, regs0, opt->min_chain_score, opt->a, rep_len, is_sr); n_regs[0] = n_regs0, regs[0] = regs0; } else { // multi-segment mm_seg_t *seg; seg = mm_seg_gen(b->km, hash, n_segs, qlens, n_regs0, regs0, n_regs, regs, a); // split fragment chain to separate segment chains free(regs0); for (i = 0; i < n_segs; ++i) { mm_set_parent(b->km, opt->mask_level, opt->mask_len, n_regs[i], regs[i], opt->a * 2 + opt->b, opt->flag&MM_F_HARD_MLEVEL, opt->alt_drop); // update mm_reg1_t::parent regs[i] = align_regs(opt, mi, b->km, qlens[i], seqs[i], &n_regs[i], regs[i], seg[i].a); mm_set_mapq(b->km, n_regs[i], regs[i], opt->min_chain_score, opt->a, rep_len, is_sr); } mm_seg_free(b->km, n_segs, seg); if (n_segs == 2 && opt->pe_ori >= 0 && (opt->flag&MM_F_CIGAR)) mm_pair(b->km, max_chain_gap_ref, opt->pe_bonus, opt->a * 2 + opt->b, opt->a, qlens, n_regs, regs); // pairing } kfree(b->km, mv.a); kfree(b->km, a); kfree(b->km, u); kfree(b->km, mini_pos); if (b->km) { km_stat(b->km, &kmst); if (mm_dbg_flag & MM_DBG_PRINT_QNAME) fprintf(stderr, "QM\t%s\t%d\tcap=%ld,nCore=%ld,largest=%ld\n", qname, qlen_sum, kmst.capacity, kmst.n_cores, kmst.largest); assert(kmst.n_blocks == kmst.n_cores); // otherwise, there is a memory leak if (kmst.largest > 1U<<28 || (opt->cap_kalloc > 0 && kmst.capacity > opt->cap_kalloc)) { if (mm_dbg_flag & MM_DBG_PRINT_QNAME) fprintf(stderr, "[W::%s] reset thread-local memory after read %s\n", __func__, qname); km_destroy(b->km); b->km = km_init(); } } } mm_reg1_t *mm_map(const mm_idx_t *mi, int qlen, const char *seq, int *n_regs, mm_tbuf_t *b, const mm_mapopt_t *opt, const char *qname) { mm_reg1_t *regs; mm_map_frag(mi, 1, &qlen, &seq, n_regs, ®s, b, opt, qname); return regs; } /************************** * Multi-threaded mapping * **************************/ typedef struct { int n_processed, n_threads, n_fp; int64_t mini_batch_size; const mm_mapopt_t *opt; mm_bseq_file_t **fp; const mm_idx_t *mi; kstring_t str; int n_parts; uint32_t *rid_shift; FILE *fp_split, **fp_parts; } pipeline_t; typedef struct { const pipeline_t *p; int n_seq, n_frag; mm_bseq1_t *seq; int *n_reg, *seg_off, *n_seg, *rep_len, *frag_gap; mm_reg1_t **reg; mm_tbuf_t **buf; } step_t; static void worker_for(void *_data, long i, int tid) // kt_for() callback { step_t *s = (step_t*)_data; int qlens[MM_MAX_SEG], j, off = s->seg_off[i], pe_ori = s->p->opt->pe_ori; const char *qseqs[MM_MAX_SEG]; double t = 0.0; mm_tbuf_t *b = s->buf[tid]; assert(s->n_seg[i] <= MM_MAX_SEG); if (mm_dbg_flag & MM_DBG_PRINT_QNAME) { fprintf(stderr, "QR\t%s\t%d\t%d\n", s->seq[off].name, tid, s->seq[off].l_seq); t = realtime(); } for (j = 0; j < s->n_seg[i]; ++j) { if (s->n_seg[i] == 2 && ((j == 0 && (pe_ori>>1&1)) || (j == 1 && (pe_ori&1)))) mm_revcomp_bseq(&s->seq[off + j]); qlens[j] = s->seq[off + j].l_seq; qseqs[j] = s->seq[off + j].seq; } if (s->p->opt->flag & MM_F_INDEPEND_SEG) { for (j = 0; j < s->n_seg[i]; ++j) { mm_map_frag(s->p->mi, 1, &qlens[j], &qseqs[j], &s->n_reg[off+j], &s->reg[off+j], b, s->p->opt, s->seq[off+j].name); s->rep_len[off + j] = b->rep_len; s->frag_gap[off + j] = b->frag_gap; } } else { mm_map_frag(s->p->mi, s->n_seg[i], qlens, qseqs, &s->n_reg[off], &s->reg[off], b, s->p->opt, s->seq[off].name); for (j = 0; j < s->n_seg[i]; ++j) { s->rep_len[off + j] = b->rep_len; s->frag_gap[off + j] = b->frag_gap; } } for (j = 0; j < s->n_seg[i]; ++j) // flip the query strand and coordinate to the original read strand if (s->n_seg[i] == 2 && ((j == 0 && (pe_ori>>1&1)) || (j == 1 && (pe_ori&1)))) { int k, t; mm_revcomp_bseq(&s->seq[off + j]); for (k = 0; k < s->n_reg[off + j]; ++k) { mm_reg1_t *r = &s->reg[off + j][k]; t = r->qs; r->qs = qlens[j] - r->qe; r->qe = qlens[j] - t; r->rev = !r->rev; } } if (mm_dbg_flag & MM_DBG_PRINT_QNAME) fprintf(stderr, "QT\t%s\t%d\t%.6f\n", s->seq[off].name, tid, realtime() - t); } static void merge_hits(step_t *s) { int f, i, k0, k, max_seg = 0, *n_reg_part, *rep_len_part, *frag_gap_part, *qlens; void *km; FILE **fp = s->p->fp_parts; const mm_mapopt_t *opt = s->p->opt; km = km_init(); for (f = 0; f < s->n_frag; ++f) max_seg = max_seg > s->n_seg[f]? max_seg : s->n_seg[f]; qlens = CALLOC(int, max_seg + s->p->n_parts * 3); n_reg_part = qlens + max_seg; rep_len_part = n_reg_part + s->p->n_parts; frag_gap_part = rep_len_part + s->p->n_parts; for (f = 0, k = k0 = 0; f < s->n_frag; ++f) { k0 = k; for (i = 0; i < s->n_seg[f]; ++i, ++k) { int j, l, t, rep_len = 0; qlens[i] = s->seq[k].l_seq; for (j = 0, s->n_reg[k] = 0; j < s->p->n_parts; ++j) { mm_err_fread(&n_reg_part[j], sizeof(int), 1, fp[j]); mm_err_fread(&rep_len_part[j], sizeof(int), 1, fp[j]); mm_err_fread(&frag_gap_part[j], sizeof(int), 1, fp[j]); s->n_reg[k] += n_reg_part[j]; if (rep_len < rep_len_part[j]) rep_len = rep_len_part[j]; } s->reg[k] = CALLOC(mm_reg1_t, s->n_reg[k]); for (j = 0, l = 0; j < s->p->n_parts; ++j) { for (t = 0; t < n_reg_part[j]; ++t, ++l) { mm_reg1_t *r = &s->reg[k][l]; uint32_t capacity; mm_err_fread(r, sizeof(mm_reg1_t), 1, fp[j]); r->rid += s->p->rid_shift[j]; if (opt->flag & MM_F_CIGAR) { mm_err_fread(&capacity, 4, 1, fp[j]); r->p = (mm_extra_t*)calloc(capacity, 4); r->p->capacity = capacity; mm_err_fread(r->p, r->p->capacity, 4, fp[j]); } } } if (!(opt->flag&MM_F_SR) && s->seq[k].l_seq >= opt->rank_min_len) mm_update_dp_max(s->seq[k].l_seq, s->n_reg[k], s->reg[k], opt->rank_frac, opt->a, opt->b); for (j = 0; j < s->n_reg[k]; ++j) { mm_reg1_t *r = &s->reg[k][j]; if (r->p) r->p->dp_max2 = 0; // reset ->dp_max2 as mm_set_parent() doesn't clear it; necessary with mm_update_dp_max() r->subsc = 0; // this may not be necessary r->n_sub = 0; // n_sub will be an underestimate as we don't see all the chains now, but it can't be accurate anyway } mm_hit_sort(km, &s->n_reg[k], s->reg[k], opt->alt_drop); mm_set_parent(km, opt->mask_level, opt->mask_len, s->n_reg[k], s->reg[k], opt->a * 2 + opt->b, opt->flag&MM_F_HARD_MLEVEL, opt->alt_drop); if (!(opt->flag & MM_F_ALL_CHAINS)) { mm_select_sub(km, opt->pri_ratio, s->p->mi->k*2, opt->best_n, 0, opt->max_gap * 0.8, &s->n_reg[k], s->reg[k]); mm_set_sam_pri(s->n_reg[k], s->reg[k]); } mm_set_mapq(km, s->n_reg[k], s->reg[k], opt->min_chain_score, opt->a, rep_len, !!(opt->flag & MM_F_SR)); } if (s->n_seg[f] == 2 && opt->pe_ori >= 0 && (opt->flag&MM_F_CIGAR)) mm_pair(km, frag_gap_part[0], opt->pe_bonus, opt->a * 2 + opt->b, opt->a, qlens, &s->n_reg[k0], &s->reg[k0]); } free(qlens); km_destroy(km); } static void *worker_pipeline(void *shared, int step, void *in) { int i, j, k; pipeline_t *p = (pipeline_t*)shared; if (step == 0) { // step 0: read sequences int with_qual = (!!(p->opt->flag & MM_F_OUT_SAM) && !(p->opt->flag & MM_F_NO_QUAL)); int with_comment = !!(p->opt->flag & MM_F_COPY_COMMENT); int frag_mode = (p->n_fp > 1 || !!(p->opt->flag & MM_F_FRAG_MODE)); step_t *s; s = (step_t*)calloc(1, sizeof(step_t)); if (p->n_fp > 1) s->seq = mm_bseq_read_frag2(p->n_fp, p->fp, p->mini_batch_size, with_qual, with_comment, &s->n_seq); else s->seq = mm_bseq_read3(p->fp[0], p->mini_batch_size, with_qual, with_comment, frag_mode, &s->n_seq); if (s->seq) { s->p = p; for (i = 0; i < s->n_seq; ++i) s->seq[i].rid = p->n_processed++; s->buf = (mm_tbuf_t**)calloc(p->n_threads, sizeof(mm_tbuf_t*)); for (i = 0; i < p->n_threads; ++i) s->buf[i] = mm_tbuf_init(); s->n_reg = (int*)calloc(5 * s->n_seq, sizeof(int)); s->seg_off = s->n_reg + s->n_seq; // seg_off, n_seg, rep_len and frag_gap are allocated together with n_reg s->n_seg = s->seg_off + s->n_seq; s->rep_len = s->n_seg + s->n_seq; s->frag_gap = s->rep_len + s->n_seq; s->reg = (mm_reg1_t**)calloc(s->n_seq, sizeof(mm_reg1_t*)); for (i = 1, j = 0; i <= s->n_seq; ++i) if (i == s->n_seq || !frag_mode || !mm_qname_same(s->seq[i-1].name, s->seq[i].name)) { s->n_seg[s->n_frag] = i - j; s->seg_off[s->n_frag++] = j; j = i; } return s; } else free(s); } else if (step == 1) { // step 1: map if (p->n_parts > 0) merge_hits((step_t*)in); else kt_for(p->n_threads, worker_for, in, ((step_t*)in)->n_frag); return in; } else if (step == 2) { // step 2: output void *km = 0; step_t *s = (step_t*)in; const mm_idx_t *mi = p->mi; for (i = 0; i < p->n_threads; ++i) mm_tbuf_destroy(s->buf[i]); free(s->buf); if ((p->opt->flag & MM_F_OUT_CS) && !(mm_dbg_flag & MM_DBG_NO_KALLOC)) km = km_init(); for (k = 0; k < s->n_frag; ++k) { int seg_st = s->seg_off[k], seg_en = s->seg_off[k] + s->n_seg[k]; for (i = seg_st; i < seg_en; ++i) { mm_bseq1_t *t = &s->seq[i]; if (p->opt->split_prefix && p->n_parts == 0) { // then write to temporary files mm_err_fwrite(&s->n_reg[i], sizeof(int), 1, p->fp_split); mm_err_fwrite(&s->rep_len[i], sizeof(int), 1, p->fp_split); mm_err_fwrite(&s->frag_gap[i], sizeof(int), 1, p->fp_split); for (j = 0; j < s->n_reg[i]; ++j) { mm_reg1_t *r = &s->reg[i][j]; mm_err_fwrite(r, sizeof(mm_reg1_t), 1, p->fp_split); if (p->opt->flag & MM_F_CIGAR) { mm_err_fwrite(&r->p->capacity, 4, 1, p->fp_split); mm_err_fwrite(r->p, r->p->capacity, 4, p->fp_split); } } } else if (s->n_reg[i] > 0) { // the query has at least one hit for (j = 0; j < s->n_reg[i]; ++j) { mm_reg1_t *r = &s->reg[i][j]; assert(!r->sam_pri || r->id == r->parent); if ((p->opt->flag & MM_F_NO_PRINT_2ND) && r->id != r->parent) continue; if (p->opt->flag & MM_F_OUT_SAM) mm_write_sam3(&p->str, mi, t, i - seg_st, j, s->n_seg[k], &s->n_reg[seg_st], (const mm_reg1_t*const*)&s->reg[seg_st], km, p->opt->flag, s->rep_len[i]); else mm_write_paf3(&p->str, mi, t, r, km, p->opt->flag, s->rep_len[i]); mm_err_puts(p->str.s); } } else if ((p->opt->flag & MM_F_PAF_NO_HIT) || ((p->opt->flag & MM_F_OUT_SAM) && !(p->opt->flag & MM_F_SAM_HIT_ONLY))) { // output an empty hit, if requested if (p->opt->flag & MM_F_OUT_SAM) mm_write_sam3(&p->str, mi, t, i - seg_st, -1, s->n_seg[k], &s->n_reg[seg_st], (const mm_reg1_t*const*)&s->reg[seg_st], km, p->opt->flag, s->rep_len[i]); else mm_write_paf3(&p->str, mi, t, 0, 0, p->opt->flag, s->rep_len[i]); mm_err_puts(p->str.s); } } for (i = seg_st; i < seg_en; ++i) { for (j = 0; j < s->n_reg[i]; ++j) free(s->reg[i][j].p); free(s->reg[i]); free(s->seq[i].seq); free(s->seq[i].name); if (s->seq[i].qual) free(s->seq[i].qual); if (s->seq[i].comment) free(s->seq[i].comment); } } free(s->reg); free(s->n_reg); free(s->seq); // seg_off, n_seg, rep_len and frag_gap were allocated with reg; no memory leak here km_destroy(km); if (mm_verbose >= 3) fprintf(stderr, "[M::%s::%.3f*%.2f] mapped %d sequences\n", __func__, realtime() - mm_realtime0, cputime() / (realtime() - mm_realtime0), s->n_seq); free(s); } return 0; } static mm_bseq_file_t **open_bseqs(int n, const char **fn) { mm_bseq_file_t **fp; int i, j; fp = (mm_bseq_file_t**)calloc(n, sizeof(mm_bseq_file_t*)); for (i = 0; i < n; ++i) { if ((fp[i] = mm_bseq_open(fn[i])) == 0) { if (mm_verbose >= 1) fprintf(stderr, "ERROR: failed to open file '%s': %s\n", fn[i], strerror(errno)); for (j = 0; j < i; ++j) mm_bseq_close(fp[j]); free(fp); return 0; } } return fp; } int mm_map_file_frag(const mm_idx_t *idx, int n_segs, const char **fn, const mm_mapopt_t *opt, int n_threads) { int i, pl_threads; pipeline_t pl; if (n_segs < 1) return -1; memset(&pl, 0, sizeof(pipeline_t)); pl.n_fp = n_segs; pl.fp = open_bseqs(pl.n_fp, fn); if (pl.fp == 0) return -1; pl.opt = opt, pl.mi = idx; pl.n_threads = n_threads > 1? n_threads : 1; pl.mini_batch_size = opt->mini_batch_size; if (opt->split_prefix) pl.fp_split = mm_split_init(opt->split_prefix, idx); pl_threads = n_threads == 1? 1 : (opt->flag&MM_F_2_IO_THREADS)? 3 : 2; kt_pipeline(pl_threads, worker_pipeline, &pl, 3); free(pl.str.s); if (pl.fp_split) fclose(pl.fp_split); for (i = 0; i < pl.n_fp; ++i) mm_bseq_close(pl.fp[i]); free(pl.fp); return 0; } int mm_map_file(const mm_idx_t *idx, const char *fn, const mm_mapopt_t *opt, int n_threads) { return mm_map_file_frag(idx, 1, &fn, opt, n_threads); } int mm_split_merge(int n_segs, const char **fn, const mm_mapopt_t *opt, int n_split_idx) { int i; pipeline_t pl; mm_idx_t *mi; if (n_segs < 1 || n_split_idx < 1) return -1; memset(&pl, 0, sizeof(pipeline_t)); pl.n_fp = n_segs; pl.fp = open_bseqs(pl.n_fp, fn); if (pl.fp == 0) return -1; pl.opt = opt; pl.mini_batch_size = opt->mini_batch_size; pl.n_parts = n_split_idx; pl.fp_parts = CALLOC(FILE*, pl.n_parts); pl.rid_shift = CALLOC(uint32_t, pl.n_parts); pl.mi = mi = mm_split_merge_prep(opt->split_prefix, n_split_idx, pl.fp_parts, pl.rid_shift); if (pl.mi == 0) { free(pl.fp_parts); free(pl.rid_shift); return -1; } for (i = n_split_idx - 1; i > 0; --i) pl.rid_shift[i] = pl.rid_shift[i - 1]; for (pl.rid_shift[0] = 0, i = 1; i < n_split_idx; ++i) pl.rid_shift[i] += pl.rid_shift[i - 1]; if (opt->flag & MM_F_OUT_SAM) for (i = 0; i < (int32_t)pl.mi->n_seq; ++i) printf("@SQ\tSN:%s\tLN:%d\n", pl.mi->seq[i].name, pl.mi->seq[i].len); kt_pipeline(2, worker_pipeline, &pl, 3); free(pl.str.s); mm_idx_destroy(mi); free(pl.rid_shift); for (i = 0; i < n_split_idx; ++i) fclose(pl.fp_parts[i]); free(pl.fp_parts); for (i = 0; i < pl.n_fp; ++i) mm_bseq_close(pl.fp[i]); free(pl.fp); mm_split_rm_tmp(opt->split_prefix, n_split_idx); return 0; }