#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->mid_occ_frac = 2e-4f; opt->sdust_thres = 0; opt->min_cnt = 3; opt->min_chain_score = 40; opt->bw = 500; opt->max_gap = 5000; opt->max_gap_ref = -1; opt->max_chain_skip = 25; 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 = 2, opt->b = 4, opt->q = 4, opt->e = 2, opt->q2 = 24, opt->e2 = 1; opt->zdrop = 400; opt->min_dp_max = opt->min_chain_score * opt->a; opt->min_ksw_len = 200; opt->mini_batch_size = 200000000; } void mm_mapopt_update(mm_mapopt_t *opt, const mm_idx_t *mi) { if (opt->flag & MM_F_SPLICE_BOTH) opt->flag &= ~(MM_F_SPLICE_FOR|MM_F_SPLICE_REV); if (opt->mid_occ <= 0) 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\n", __func__, realtime() - mm_realtime0, cputime() / (realtime() - mm_realtime0), opt->mid_occ); } int mm_set_opt(const char *preset, mm_idxopt_t *io, mm_mapopt_t *mo) { if (preset == 0) { mm_idxopt_init(io); mm_mapopt_init(mo); } else if (strcmp(preset, "ava-ont") == 0) { io->is_hpc = 0, io->k = 15, io->w = 5; mo->flag |= MM_F_AVA | MM_F_NO_SELF; mo->min_chain_score = 100, mo->pri_ratio = 0.0f, mo->max_gap = 10000, mo->max_chain_skip = 25; mo->mini_batch_size = 500000000; } else if (strcmp(preset, "ava-pb") == 0) { io->is_hpc = 1, io->k = 19, io->w = 5; mo->flag |= MM_F_AVA | MM_F_NO_SELF; mo->min_chain_score = 100, mo->pri_ratio = 0.0f, mo->max_gap = 10000, mo->max_chain_skip = 25; mo->mini_batch_size = 500000000; } else if (strcmp(preset, "map10k") == 0 || strcmp(preset, "map-pb") == 0) { io->is_hpc = 1, io->k = 19; } else if (strcmp(preset, "map-ont") == 0) { io->is_hpc = 0, io->k = 15; } else if (strcmp(preset, "asm5") == 0) { io->is_hpc = 0, io->k = 19, io->w = 19; mo->a = 1, mo->b = 19, mo->q = 39, mo->q2 = 81, mo->e = 3, mo->e2 = 1, mo->zdrop = 200; mo->min_dp_max = 200; } else if (strcmp(preset, "asm10") == 0) { io->is_hpc = 0, io->k = 19, io->w = 19; mo->a = 1, mo->b = 9, mo->q = 16, mo->q2 = 41, mo->e = 2, mo->e2 = 1, mo->zdrop = 200; mo->min_dp_max = 200; } else if (strcmp(preset, "short") == 0 || strcmp(preset, "sr") == 0) { io->is_hpc = 0, io->k = 21, io->w = 11; mo->flag |= MM_F_APPROX_EXT; mo->a = 2, mo->b = 8, mo->q = 12, mo->e = 2, mo->q2 = 32, mo->e2 = 1; mo->max_gap = 100; mo->pri_ratio = 0.5f; mo->min_cnt = 2; mo->min_chain_score = 20; mo->min_dp_max = 40; mo->best_n = 20; mo->bw = 50; mo->mid_occ = 1000; mo->mini_batch_size = 50000000; } else if (strcmp(preset, "splice") == 0 || strcmp(preset, "cdna") == 0) { io->is_hpc = 0, io->k = 15, io->w = 5; mo->flag |= MM_F_SPLICE | MM_F_SPLICE_FOR | MM_F_SPLICE_REV; mo->max_gap = 2000, mo->max_gap_ref = mo->bw = 200000; mo->a = 1, mo->b = 2, mo->q = 2, mo->e = 1, mo->q2 = 32, mo->e2 = 0; mo->noncan = 5; mo->zdrop = 200; } else return -1; return 0; } typedef struct { uint32_t n; 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_dbg_flag & 1)) 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; } 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) { int i, n, j, n_u, max_gap_ref, rep_st = 0, rep_en = 0, rep_len = 0; int64_t n_a; uint64_t *u; mm_match_t *m; mm128_t *a; mm_reg1_t *regs; // collect minimizers b->mini.n = 0; mm_sketch(b->km, seq, qlen, mi->w, mi->k, 0, mi->is_hpc, &b->mini); n = b->mini.n; if (opt->sdust_thres > 0) mm_dust_minier(&b->mini, qlen, seq, opt->sdust_thres, b->sdb); // 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[i].qpos = (uint32_t)p->y; m[i].x.cr = mm_idx_get(mi, p->x>>8, &t); m[i].n = t; } // 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]; mm_match_t *q = &m[i]; const uint64_t *r = q->x.cr; int k, q_span = p->x & 0xff, is_tandem = 0; if (q->n >= opt->mid_occ) { int en = (q->qpos>>1) + 1, st = en - q_span; if (st > rep_en) { rep_len += rep_en - rep_st; rep_st = st, rep_en = en; } else rep_en = en; continue; } if (i > 0 && p->x>>8 == b->mini.a[i - 1].x>>8) is_tandem = 1; if (i < n - 1 && p->x>>8 == b->mini.a[i + 1].x>>8) is_tandem = 1; for (k = 0; k < q->n; ++k) { int32_t rpos = (uint32_t)r[k] >> 1; mm128_t *p; if (qname && (opt->flag&(MM_F_NO_SELF|MM_F_AVA))) { const char *tname = mi->seq[r[k]>>32].name; if ((opt->flag&MM_F_NO_SELF) && strcmp(qname, tname) == 0 && rpos == (q->qpos>>1)) // avoid the diagonal continue; if ((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) | rpos; p->y = (uint64_t)q_span << 32 | q->qpos >> 1; } else { // reverse strand p->x = 1ULL<<63 | (r[k]&0xffffffff00000000ULL) | rpos; p->y = (uint64_t)q_span << 32 | (qlen - ((q->qpos>>1) + 1 - q_span) - 1); } if (is_tandem) p->y |= MM_SEED_TANDEM; } } rep_len += rep_en - rep_st; n_a = j; radix_sort_128x(a, a + n_a); kfree(b->km, m); 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)); } max_gap_ref = opt->max_gap_ref >= 0? opt->max_gap_ref : opt->max_gap; n_u = mm_chain_dp(max_gap_ref, opt->max_gap, opt->bw, opt->max_chain_skip, opt->min_cnt, opt->min_chain_score, !!(opt->flag&MM_F_SPLICE), n_a, a, &u, b->km); regs = mm_gen_regs(b->km, qlen, n_u, u, a); *n_regs = n_u; if (mm_dbg_flag & MM_DBG_PRINT_SEED) for (j = 0; j < n_u; ++j) for (i = regs[j].as; i < regs[j].as + regs[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 == regs[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)); 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, opt->a * 2 + opt->b); mm_select_sub(b->km, opt->mask_level, opt->pri_ratio, mi->k*2, opt->best_n, n_regs, regs); if (!(opt->flag & MM_F_SPLICE)) 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_set_parent(b->km, opt->mask_level, *n_regs, regs, opt->a * 2 + opt->b); mm_select_sub(b->km, opt->mask_level, opt->pri_ratio, mi->k*2, opt->best_n, n_regs, regs); mm_set_sam_pri(*n_regs, regs); } } mm_set_mapq(*n_regs, regs, opt->min_chain_score, opt->a, rep_len); // free kfree(b->km, a); kfree(b->km, u); return regs; } /************************** * Multi-threaded mapping * **************************/ typedef struct { int mini_batch_size, n_processed, n_threads; const mm_mapopt_t *opt; mm_bseq_file_t *fp; const mm_idx_t *mi; kstring_t str; } pipeline_t; typedef struct { const pipeline_t *p; int n_seq; mm_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; if (mm_dbg_flag & MM_DBG_PRINT_QNAME) fprintf(stderr, "QR\t%s\t%d\n", step->seq[i].name, tid); 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 int with_qual = (!!(p->opt->flag & MM_F_OUT_SAM) && !(p->opt->flag & MM_F_NO_QUAL)); step_t *s; s = (step_t*)calloc(1, sizeof(step_t)); s->seq = mm_bseq_read(p->fp, p->mini_batch_size, with_qual, &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 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 (i = 0; i < s->n_seq; ++i) { mm_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, s->n_reg[i], s->reg[i]); else mm_write_paf(&p->str, mi, t, r, km, p->opt->flag); puts(p->str.s); } if (s->n_reg[i] == 0 && (p->opt->flag & MM_F_OUT_SAM)) { mm_write_sam(&p->str, 0, t, 0, 0, 0); puts(p->str.s); } 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); } free(s->reg); free(s->n_reg); free(s->seq); 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; } int mm_map_file(const mm_idx_t *idx, const char *fn, const mm_mapopt_t *opt, int n_threads) { pipeline_t pl; memset(&pl, 0, sizeof(pipeline_t)); pl.fp = mm_bseq_open(fn); if (pl.fp == 0) { if (mm_verbose >= 1) fprintf(stderr, "ERROR: failed to open file '%s'\n", fn); return -1; } pl.opt = opt, pl.mi = idx; pl.n_threads = n_threads, pl.mini_batch_size = opt->mini_batch_size; if ((opt->flag & MM_F_OUT_SAM) && !(opt->flag & MM_F_NO_SAM_SQ)) mm_write_sam_SQ(idx); kt_pipeline(n_threads == 1? 1 : 2, worker_pipeline, &pl, 3); free(pl.str.s); mm_bseq_close(pl.fp); return 0; }