#include #include #include #include "kthread.h" #include "kvec.h" #include "kalloc.h" #include "sdust.h" #include "mmpriv.h" #include "bseq.h" void mm_mapopt_init(mm_mapopt_t *opt) { memset(opt, 0, sizeof(mm_mapopt_t)); opt->max_occ_frac = 1e-5f; opt->mid_occ_frac = 2e-4f; opt->sdust_thres = 0; opt->min_cnt = 3; opt->min_chain_score = 40; opt->bw = 1000; opt->max_gap = 10000; opt->max_chain_skip = 15; opt->min_seedcov_ratio = 0.0f; opt->mask_level = 0.5f; opt->pri_ratio = 0.8f; opt->best_n = 5; opt->max_join_long = 20000; opt->max_join_short = 2000; opt->min_join_flank_sc = 1000; opt->a = 1, opt->b = 2, opt->q = 2, opt->e = 1; opt->zdrop = 200; opt->min_dp_max = opt->min_chain_score; opt->min_ksw_len = 100; } void mm_mapopt_update(mm_mapopt_t *opt, const mm_idx_t *mi) { opt->max_occ = mm_idx_cal_max_occ(mi, opt->max_occ_frac); opt->mid_occ = mm_idx_cal_max_occ(mi, opt->mid_occ_frac); if (mm_verbose >= 3) fprintf(stderr, "[M::%s::%.3f*%.2f] mid_occ = %d; max_occ = %d\n", __func__, realtime() - mm_realtime0, cputime() / (realtime() - mm_realtime0), opt->mid_occ, opt->max_occ); } typedef struct { uint32_t n:31, is_alloc:1; uint32_t qpos; union { const uint64_t *cr; uint64_t *r; } x; } mm_match_t; struct mm_tbuf_s { sdust_buf_t *sdb; mm128_v mini; void *km; }; mm_tbuf_t *mm_tbuf_init(void) { mm_tbuf_t *b; b = (mm_tbuf_t*)calloc(1, sizeof(mm_tbuf_t)); if (mm_verbose < 10) b->km = km_init(); b->sdb = sdust_buf_init(b->km); return b; } void mm_tbuf_destroy(mm_tbuf_t *b) { if (b == 0) return; kfree(b->km, b->mini.a); sdust_buf_destroy(b->sdb); km_destroy(b->km); free(b); } static void mm_dust_minier(mm128_v *mini, int l_seq, const char *seq, int sdust_thres, sdust_buf_t *sdb) { int n_dreg, j, k, u = 0; const uint64_t *dreg; if (sdust_thres <= 0 || sdb == 0) return; dreg = sdust_core((const uint8_t*)seq, l_seq, sdust_thres, 64, &n_dreg, sdb); for (j = k = 0; j < mini->n; ++j) { // squeeze out minimizers that significantly overlap with LCRs int32_t qpos = (uint32_t)mini->a[j].y>>1, span = mini->a[j].x&0xff; int32_t s = qpos - (span - 1), e = s + span; while (u < n_dreg && (uint32_t)dreg[u] <= s) ++u; if (u < n_dreg && dreg[u]>>32 < e) { int v, l = 0; for (v = u; v < n_dreg && dreg[v]>>32 < e; ++v) { // iterate over LCRs overlapping this minimizer int ss = s > dreg[v]>>32? s : dreg[v]>>32; int ee = e < (uint32_t)dreg[v]? e : (uint32_t)dreg[v]; l += ee - ss; } if (l <= span>>1) mini->a[k++] = mini->a[j]; // keep the minimizer if less than half of it falls in masked region } } mini->n = k; } #if 0 int mm_pair_thin_core(mm_tbuf_t *b, uint64_t x, int radius, int rel, int st0, int n, const uint64_t *z, uint64_v *a) { int i, st = st0, en = n, mid = en - 1; while (st < en) { uint64_t y; mid = st + ((en - st) >> 1); y = z[mid]; if (y < x && (x - y)>>1 > radius) st = mid + 1; else if (y >= x && (y - x)>>1 > radius) en = mid; else break; } if (st < en) { for (en = mid + 1; en < n; ++en) if (z[en] > x && (z[en] - x)>>1 > radius) break; for (st = mid - 1; st >= st0; --st) if (z[st] < x && (x - z[st])>>1 > radius) break; ++st; for (i = st; i < en; ++i) { uint64_t y = z[i]; if (((x ^ y) & 1) == rel) { // printf("* %d,%d\n", (uint32_t)x>>1, (uint32_t)y>>1); kv_push(uint64_t, b->km, *a, y); } } return en; } else return st < n && z[st] < x? st + 1 : en; } void mm_pair_thin(mm_tbuf_t *b, int radius, mm_match_t *m1, mm_match_t *m2) { mm_match_t *m[2]; const uint64_t *z[2]; uint64_v a[2]; int i, n[2], k[2], u = 0, rel = (m1->qpos ^ m2->qpos) & 1; m[0] = m1, m[1] = m2; for (i = 0; i < 2; ++i) { n[i] = m[i]->n; z[i] = m[i]->x.cr; k[i] = 0; kv_init(a[i]); kv_resize(uint64_t, b->km, a[i], 256); } while (k[0] < n[0] && k[1] < n[1]) { //printf("%d; %d,%d\n", u, k[0], k[1]); int v = u^1, dist = (int)(m[v]->qpos>>1) - (int)(m[u]->qpos>>1); uint64_t x = z[u][k[u]]; int uori = (x ^ m[u]->qpos) & 1, last; int64_t tpos = x>>1 & 0x7fffffff; tpos = uori == 0? tpos + dist : tpos - dist; if (tpos < 0) tpos = 0; x = x>>32<<32 | tpos<<1 | (x&1); last = a[v].n; k[v] = mm_pair_thin_core(b, x, radius, rel, k[v], n[v], z[v], &a[v]); if (a[v].n > last) kv_push(uint64_t, b->km, a[u], z[u][k[u]]); ++k[u]; u ^= 1; } for (i = 0; i < 2; ++i) m[i]->n = a[i].n, m[i]->x.r = a[i].a, m[i]->is_alloc = 1; // printf("%d,%d; %d,%d\n", m[0]->qpos>>1, m[1]->qpos>>1, m[0]->n, m[1]->n); } #endif mm_reg1_t *mm_map_frag(const mm_mapopt_t *opt, const mm_idx_t *mi, mm_tbuf_t *b, uint32_t m_st, uint32_t m_en, const char *qname, int qlen, const char *seq, int *n_regs) { int i, n = m_en - m_st, j, n_u; int64_t n_a; uint64_t *u; mm_match_t *m; mm128_t *a; mm_reg1_t *regs; // convert to local representation m = (mm_match_t*)kmalloc(b->km, n * sizeof(mm_match_t)); for (i = 0; i < n; ++i) { int t; mm128_t *p = &b->mini.a[i + m_st]; m[i].is_alloc = 0; m[i].qpos = (uint32_t)p->y; m[i].x.cr = mm_idx_get(mi, p->x>>8, &t); m[i].n = t; } #if 0 int last = -1, last2 = -1; // pair k-mer thinning for (i = 0; i < n; ++i) { if (m[i].n >= opt->mid_occ && m[i].n < opt->max_occ) { if (last2 < 0) last2 = i; if (last < 0 || m[last].n < m[i].n) last = i; if (last >= 0 && (m[last].qpos>>1) + (m[last].span>>1) <= m[i].qpos>>1) { mm_pair_thin(b, opt->bw, &m[last], &m[i]); last2 = last = -1; } else if (last2 >= 0 && (m[last2].qpos>>1) + (m[last2].span>>1) <= m[i].qpos>>1) { mm_pair_thin(b, opt->bw, &m[last2], &m[i]); last2 = last = -1; } } } #endif // fill the _a_ array for (i = 0, n_a = 0; i < n; ++i) // find the length of a[] if (m[i].n < opt->mid_occ) n_a += m[i].n; a = (mm128_t*)kmalloc(b->km, n_a * sizeof(mm128_t)); for (i = j = 0; i < n; ++i) { mm128_t *p = &b->mini.a[i + m_st]; mm_match_t *q = &m[i]; const uint64_t *r = q->x.cr; int k, q_span = p->x & 0xff; if (q->n >= opt->mid_occ) continue; for (k = 0; k < q->n; ++k) { const char *tname = mi->seq[r[k]>>32].name; int32_t rpos = (uint32_t)r[k] >> 1; mm128_t *p; if (qname && (opt->flag&MM_F_NO_SELF) && strcmp(qname, tname) == 0 && rpos == (q->qpos>>1)) // avoid the diagonal continue; if (qname && (opt->flag&MM_F_AVA) && strcmp(qname, tname) > 0) // all-vs-all mode: map once continue; p = &a[j++]; if ((r[k]&1) == (q->qpos&1)) { // forward strand p->x = (r[k]&0xffffffff00000000ULL) | (uint32_t)r[k]>>1; p->y = (uint64_t)q_span << 32 | q->qpos >> 1; } else { // reverse strand p->x = 1ULL<<63 | (r[k]&0xffffffff00000000ULL) | (uint32_t)r[k]>>1; p->y = (uint64_t)q_span << 32 | (qlen - ((q->qpos>>1) + 1 - q_span) - 1); } } } n_a = j; radix_sort_128x(a, a + n_a); for (i = 0; i < n; ++i) if (m[i].is_alloc) kfree(b->km, m[i].x.r); kfree(b->km, m); n_u = mm_chain_dp(opt->max_gap, opt->bw, opt->max_chain_skip, opt->min_cnt, opt->min_chain_score, n_a, a, &u, b->km); regs = mm_gen_regs(b->km, qlen, n_u, u, a); *n_regs = n_u; if (!(opt->flag & MM_F_AVA)) { // don't choose primary mapping(s) for read overlap mm_set_parent(b->km, opt->mask_level, *n_regs, regs); mm_select_sub(b->km, opt->mask_level, opt->pri_ratio, opt->best_n, n_regs, regs); mm_join_long(b->km, opt, qlen, n_regs, regs, a); // TODO: this can be applied to all-vs-all in principle } if (opt->flag & MM_F_CIGAR) { regs = mm_align_skeleton(b->km, opt, mi, qlen, seq, n_regs, regs, a); // this calls mm_filter_regs() if (!(opt->flag & MM_F_AVA)) { mm_update_parent(b->km, opt->mask_level, *n_regs, regs); mm_select_sub(b->km, opt->mask_level, opt->pri_ratio, opt->best_n, n_regs, regs); } } else mm_filter_regs(b->km, opt, n_regs, regs); mm_set_mapq(*n_regs, regs); // free kfree(b->km, a); kfree(b->km, u); return regs; } mm_reg1_t *mm_map(const mm_idx_t *mi, int l_seq, const char *seq, int *n_regs, mm_tbuf_t *b, const mm_mapopt_t *opt, const char *qname) { mm_reg1_t *regs; if (mm_verbose >= 11) fprintf(stderr, "===> %s <===\n", qname); b->mini.n = 0; mm_sketch(b->km, seq, l_seq, mi->w, mi->k, 0, mi->is_hpc, &b->mini); if (opt->sdust_thres > 0) mm_dust_minier(&b->mini, l_seq, seq, opt->sdust_thres, b->sdb); regs = mm_map_frag(opt, mi, b, 0, b->mini.n, qname, l_seq, seq, n_regs); return regs; } /************************** * Multi-threaded mapping * **************************/ typedef struct { int mini_batch_size, n_processed, n_threads; const mm_mapopt_t *opt; bseq_file_t *fp; const mm_idx_t *mi; kstring_t str; } pipeline_t; typedef struct { const pipeline_t *p; int n_seq; bseq1_t *seq; int *n_reg; 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 *step = (step_t*)_data; step->reg[i] = mm_map(step->p->mi, step->seq[i].l_seq, step->seq[i].seq, &step->n_reg[i], step->buf[tid], step->p->opt, step->seq[i].name); } static void *worker_pipeline(void *shared, int step, void *in) { int i, j; pipeline_t *p = (pipeline_t*)shared; if (step == 0) { // step 0: read sequences step_t *s; s = (step_t*)calloc(1, sizeof(step_t)); s->seq = bseq_read(p->fp, p->mini_batch_size, !!(p->opt->flag & MM_F_OUT_SAM), &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(s->n_seq, sizeof(int)); s->reg = (mm_reg1_t**)calloc(s->n_seq, sizeof(mm_reg1_t*)); return s; } else free(s); } else if (step == 1) { // step 1: map kt_for(p->n_threads, worker_for, in, ((step_t*)in)->n_seq); return in; } else if (step == 2) { // step 2: output 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); for (i = 0; i < s->n_seq; ++i) { bseq1_t *t = &s->seq[i]; for (j = 0; j < s->n_reg[i]; ++j) { mm_reg1_t *r = &s->reg[i][j]; if (p->opt->flag & MM_F_OUT_SAM) mm_write_sam(&p->str, mi, t, r); else mm_write_paf(&p->str, mi, t, r); puts(p->str.s); free(r->p); } if (s->n_reg[i] == 0 && (p->opt->flag & MM_F_OUT_SAM)) { mm_write_sam(&p->str, 0, t, 0); puts(p->str.s); } free(s->reg[i]); free(s->seq[i].seq); free(s->seq[i].name); if (s->seq[i].qual) free(s->seq[i].qual); } free(s->reg); free(s->n_reg); free(s->seq); 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; } int mm_map_file(const mm_idx_t *idx, const char *fn, const mm_mapopt_t *opt, int n_threads, int mini_batch_size) { pipeline_t pl; memset(&pl, 0, sizeof(pipeline_t)); pl.fp = bseq_open(fn); if (pl.fp == 0) return -1; pl.opt = opt, pl.mi = idx; pl.n_threads = n_threads, pl.mini_batch_size = mini_batch_size; if (opt->flag & MM_F_OUT_SAM) { uint32_t i; for (i = 0; i < idx->n_seq; ++i) printf("@SQ\tSN:%s\tLN:%d\n", idx->seq[i].name, idx->seq[i].len); } kt_pipeline(n_threads == 1? 1 : 2, worker_pipeline, &pl, 3); free(pl.str.s); bseq_close(pl.fp); return 0; }