#include #include #include #include #include #include "kseq.h" // FASTA/Q parser #include "kavl.h" #include "khash.h" #include "kalloc.h" #include "kthread.h" #include "inter.h" #include "Overlaps.h" #include "CommandLines.h" #include "htab.h" #include "Hash_Table.h" #include "Correct.h" #include "Process_Read.h" #include "Assembly.h" KSEQ_INIT(gzFile, gzread) void ha_get_ul_candidates_interface(ha_abufl_t *ab, int64_t rid, char* rs, uint64_t rl, uint64_t mz_w, uint64_t mz_k, const ul_idx_t *uref, overlap_region_alloc *overlap_list, overlap_region_alloc *overlap_list_hp, Candidates_list *cl, double bw_thres, int max_n_chain, int keep_whole_chain, kvec_t_u8_warp* k_flag, kvec_t_u64_warp* chain_idx, overlap_region* f_cigar, kvec_t_u64_warp* dbg_ct, st_mt_t *sp, void *km); #define G_CHAIN_BW 16//128 #define FLANK_M (0x7fffU) #define P_CHAIN_COV 0.985 #define P_FRAGEMENT_CHAIN_COV 0.20 #define P_FRAGEMENT_PRIMARY_CHAIN_COV 0.70 #define P_FRAGEMENT_PRIMARY_SECOND_COV 0.25 #define P_CHAIN_SCORE 0.6 #define G_CHAIN_GAP 0.1 #define UG_SKIP 5 #define RG_SKIP 25 #define G_CHAIN_TRANS_RATE 0.25 #define G_CHAIN_TRANS_WEIGHT -1 #define G_CHAIN_INDEL 128 #define W_CHN_PEN_GAP 0.1 #define N_GCHAIN_RATE 0.04 #define MG_SEED_IGNORE (1ULL<<41) #define MG_SEED_TANDEM (1ULL<<42) #define MG_SEED_KEPT (1ULL<<43) #define MG_MAX_SEG 255 #define MG_SEED_SEG_SHIFT 48 #define MG_SEED_SEG_MASK (0xffULL<<(MG_SEED_SEG_SHIFT)) #define mg_seg_id(a) ((int32_t)(((a).y&MG_SEED_SEG_MASK) >> MG_SEED_SEG_SHIFT)) #define MG_SEED_WT_SHIFT 56 #define MG_MAX_SHORT_K 15 #define MG_SHORT_K_EXT 10000 ///1000 in minigraph #define generic_key(x) (x) KRADIX_SORT_INIT(gfa64, uint64_t, generic_key, 8) #define ul_ov_srt_qe_key(p) ((p).qe) KRADIX_SORT_INIT(ul_ov_srt_qe, ul_ov_t, ul_ov_srt_qe_key, member_size(ul_ov_t, qe)) #define ul_ov_srt_qs_key(p) ((p).qs) KRADIX_SORT_INIT(ul_ov_srt_qs, ul_ov_t, ul_ov_srt_qs_key, member_size(ul_ov_t, qs)) #define ul_ov_srt_tn_key(p) ((p).tn) KRADIX_SORT_INIT(ul_ov_srt_tn, ul_ov_t, ul_ov_srt_tn_key, member_size(ul_ov_t, tn)) #define ul_ov_srt_qn_key(p) ((p).qn) KRADIX_SORT_INIT(ul_ov_srt_qn, ul_ov_t, ul_ov_srt_qn_key, member_size(ul_ov_t, qn)) #define utg_ct_t_x_key(p) ((p).x) KRADIX_SORT_INIT(utg_ct_t_x_srt, utg_ct_t, utg_ct_t_x_key, member_size(utg_ct_t, x)) #define utg_ct_t_s_key(p) ((p).s) KRADIX_SORT_INIT(utg_ct_t_s_srt, utg_ct_t, utg_ct_t_s_key, member_size(utg_ct_t, s)) #define hap_ev_cov_key(x) ((x).cov) KRADIX_SORT_INIT(hap_ev_cov_srt, haplotype_evdience, hap_ev_cov_key, member_size(haplotype_evdience, cov)) struct mg_tbuf_s { void *km; int frag_gap; }; typedef struct mg_tbuf_s mg_tbuf_t; mg_tbuf_t *mg_tbuf_init(void) { mg_tbuf_t *b; b = (mg_tbuf_t*)calloc(1, sizeof(mg_tbuf_t)); b->km = km_init(); return b; } void mg_tbuf_destroy(mg_tbuf_t *b) { if (b == 0) return; if (b->km) km_destroy(b->km); free(b); } void *mg_tbuf_get_km(mg_tbuf_t *b) { return b->km; } typedef struct { int w, k, bw, max_gap, is_HPC, hap_n, occ_weight, max_gap_pre, max_gc_seq_ext, seed; int max_lc_skip, max_lc_iter, min_lc_cnt, min_lc_score, max_gc_skip, ref_bonus; int min_gc_cnt, min_gc_score, sub_diff, best_n; float chn_pen_gap, mask_level, pri_ratio; ///base-alignment double bw_thres, diff_ec_ul; int max_n_chain; } mg_idxopt_t; typedef struct { ///off: start idx in mg128_t * a[]; ///cnt: how many eles in this chain ///a[off, off+cnt) saves the eles in this chain int32_t off, cnt:31, inner_pre:1; ///ref_id|rev uint32_t v; ///chain in ref: [rs, re) ///chain in query: [qs, qe) int32_t rs, re, qs, qe; ///score: chain score int32_t score, dist_pre; uint32_t hash_pre; } mg_lchain_t; typedef struct { uint32_t v, d; int32_t pre; } mg_pathv_t; typedef struct { int32_t qs, qe, rs, re; uint32_t v; } mg_coor_t; ///mg128_t->y: weight(8)seg_id(8)flag(8)span(8)pos(32) ///mg128_t->x: rid(31)rev(1)pos(33); keep reference typedef struct { uint64_t x, y; } mg128_t; #define sort_key_128x(a) ((a).x) KRADIX_SORT_INIT(128x, mg128_t, sort_key_128x, 8) void radix_sort_128x(mg128_t *beg, mg128_t *end); typedef struct { int32_t off, cnt; uint32_t v; int32_t score; } mg_llchain_t; typedef struct { int32_t id, parent; int32_t off, cnt; int32_t n_anchor, score; int32_t qs, qe; int32_t plen, ps, pe; int32_t blen, mlen; float div; uint32_t hash; int32_t subsc, n_sub; uint32_t mapq:8, flt:1, dummy:23; } mg_gchain_t; typedef struct { size_t n,m; uint64_t *a, tl; kvec_t(char) cc; } mg_dbn_t; typedef struct { int32_t cnt; uint32_t v; int32_t score; uint32_t qs, qe, ts, te; } mg_lres_t; typedef struct { int32_t n_gc, n_lc; mg_gchain_t *gc;///g_chain; idx in l_chains mg_lres_t *lc;///l_chain uint64_t qid, qlen; } mg_gres_t; typedef struct { size_t n,m; mg_gres_t *a; uint64_t total_base; uint64_t total_pair; } mg_gres_a; typedef struct { // global data structure for kt_pipeline() const void *ha_flt_tab; const ha_pt_t *ha_idx; const mg_idxopt_t *opt; const ma_ug_t *ug; const asg_t *rg; const ug_opt_t *uopt; const ul_idx_t *uu; kseq_t *ks; int64_t chunk_size; uint64_t n_thread; uint64_t total_base; uint64_t total_pair; mg_gres_a hits; mg_dbn_t nn; uint64_t num_bases, num_corrected_bases, num_recorrected_bases; } uldat_t; typedef struct { uint64_t asm_size; uint64_t asm_cov; } mul_ov_t; ///three levels: ///level-0: minimizers ///level-1: linear chains ///level-2: g chains ///gc[] saves the idx in lc[], lc saves the idx in a[] typedef struct { void *km; int32_t n_gc, n_lc, n_a, rep_len; mg_gchain_t *gc;///g_chain; idx in l_chains mg_llchain_t *lc;///l_chain mg128_t *a; // minimizer positions; see comments above mg_update_anchors() for details uint64_t qid, qlen; } mg_gchains_t; typedef struct { uint32_t n; ///length of candidate list uint64_t q_span:31, rev:1, q_pos:32; uint32_t qid:16, weight:15, is_tandem:1; const ha_idxposl_t *cr; ///candidate list } mg_match_t; typedef struct { uint64_t qse, rse, gld; } lc_srt_t; #define lc_srt_key(p) ((p).qse) KRADIX_SORT_INIT(lc_srt, lc_srt_t, lc_srt_key, member_size(lc_srt_t, qse)) typedef struct { uint64_t x, e; int32_t d; uint32_t id; } eg_srt_t; #define eg_srt_x_key(p) ((p).x) KRADIX_SORT_INIT(eg_srt_x, eg_srt_t, eg_srt_x_key, member_size(eg_srt_t, x)) #define eg_srt_d_key(p) ((p).d) KRADIX_SORT_INIT(eg_srt_d, eg_srt_t, eg_srt_d_key, member_size(eg_srt_t, d)) // shortest path typedef struct { // input ///(lj_ref_id)|(lj_ref_rev^1) uint32_t v; ///target_dist should like the overlap length in string graph ///it should be used to evaluate if the identified path is close to real path/alignment int32_t target_dist; uint32_t target_hash; ///inner: if li and lj are at the same ref id ///meta: j uint32_t meta:30, check_hash:1, inner:1; /** * There are two cases: * (1) lj->qs************lj->qe * li->qs************li->qe * (2) lj->qs************lj->qe * li->qs************li->qe * qlen = li->qs - lj->qe;///might be negative * **/ int32_t qlen/**, so**/; // output uint32_t n_path:31, is_0:1;///I guess n_path is how many path from src to dest int32_t path_end;///looks like an idx to alignment int32_t dist, mlen; uint32_t hash; // aux uint64_t srt_key; } mg_path_dst_t; typedef struct { uint32_t srt; int32_t i; } gc_frag_t; ///I think this structure is just used for iteration ///iterate each ref id, instead of alignment id typedef struct sp_node_s { uint64_t di; // dist<<32 | node_id in avl tree(doesn't matter too much) uint32_t v;///ref_id|rev int32_t pre; uint32_t hash;///hash is path hash, instead of node hash int32_t is_0; KAVL_HEAD(struct sp_node_s) head; } sp_node_t, *sp_node_p; typedef struct { int32_t k, mlen;//k: number of walks from src to this node int32_t qs, qe; sp_node_t *p[MG_MAX_SHORT_K]; // this forms a max-heap } sp_topk_t; #define gc_frag_key(p) ((p).srt) KRADIX_SORT_INIT(gc, gc_frag_t, gc_frag_key, 4) #define dst_key(p) ((p).srt_key) KRADIX_SORT_INIT(dst, mg_path_dst_t, dst_key, 8) #define sp_node_cmp(a, b) (((a)->di > (b)->di) - ((a)->di < (b)->di)) KAVL_INIT(sp, sp_node_t, head, sp_node_cmp) #define sp_node_lt(a, b) ((a)->di < (b)->di) KSORT_INIT(sp, sp_node_p, sp_node_lt) KHASH_MAP_INIT_INT(sp, sp_topk_t) KHASH_MAP_INIT_INT(sp2, uint64_t) typedef struct { kv_ul_ov_t lo; kv_ul_ov_t tk; kvec_t_u64_warp srt; }glchain_t; typedef struct { mg_lchain_t *a; size_t n, m; }vec_mg_lchain_t; typedef struct { mg_path_dst_t *a; size_t n, m; }vec_mg_path_dst_t; typedef struct { sp_node_t **a; size_t n, m; }vec_sp_node_t; typedef struct { mg_pathv_t *a; size_t n, m; }vec_mg_pathv_t; typedef struct { vec_mg_lchain_t l; vec_mg_lchain_t swap; vec_mg_path_dst_t dst; vec_sp_node_t out; vec_mg_pathv_t path; kvec_t(uint64_t) v; kvec_t(int64_t) f; st_mt_t dst_done; }gdpchain_t; typedef struct { // data structure for each step in kt_pipeline() const mg_idxopt_t *opt; const void *ha_flt_tab; const ha_pt_t *ha_idx; const ma_ug_t *ug; const asg_t *rg; const ug_opt_t *uopt; const ul_idx_t *uu; int n, m, sum_len; uint64_t *len, id; char **seq; ha_mzl_v *mzs;///useless st_mt_t *sps;///useless mg_gchains_t **gcs;///useless mg_tbuf_t **buf;///useless ha_ovec_buf_t **hab; glchain_t *ll; gdpchain_t *gdp; // glchain_t *sec_ll; uint64_t num_bases, num_corrected_bases, num_recorrected_bases; } utepdat_t; void hc_glchain_destroy(glchain_t *b) { if (!b) return; kv_destroy(b->lo); kv_destroy(b->tk); kv_destroy(b->srt.a); } void hc_gdpchain_destroy(gdpchain_t *b) { if (!b) return; kv_destroy(b->l); kv_destroy(b->swap); kv_destroy(b->dst); kv_destroy(b->out); kv_destroy(b->path); kv_destroy(b->v); kv_destroy(b->f); kv_destroy(b->dst_done); } void init_mg_opt(mg_idxopt_t *opt, int is_HPC, int k, int w, int hap_n, int max_n_chain, double bw_thres, double diff_ec_ul) { opt->k = k; opt->w = w; opt->hap_n = hap_n; opt->is_HPC = is_HPC; opt->bw = 10000;///2000 in minigraph opt->max_gap = 500000;///5000 in minigraph opt->occ_weight = 20; opt->max_gap_pre = 10000;///1000 in minigraph opt->max_lc_iter = 10000; opt->chn_pen_gap = 0.19;///using minimap2's value opt->max_lc_skip = 25;// mo->max_gc_skip = 25; opt->max_lc_iter = 10000; opt->min_lc_cnt = 2; opt->min_lc_score = 30; opt->max_gc_skip = 25; opt->ref_bonus = 0; opt->mask_level = 0.5f; opt->max_gc_seq_ext = 5; opt->seed = 11; opt->min_gc_cnt = 3, opt->min_gc_score = 50; opt->sub_diff = 6; opt->best_n = 5; opt->pri_ratio = 0.8f; opt->max_n_chain = max_n_chain; opt->bw_thres = bw_thres; opt->diff_ec_ul = diff_ec_ul; } void uidx_l_build(ma_ug_t *ug, mg_idxopt_t *opt, int cutoff) { ha_flt_tab = ha_ft_ul_gen(&asm_opt, &(ug->u), opt->k, opt->w, cutoff); ha_idx = ha_pt_ul_gen(&asm_opt, ha_flt_tab, &(ug->u), opt->k, opt->w, cutoff); fprintf(stderr, "[M::%s] Index has been built.\n", __func__); } void uidx_build(ma_ug_t *ug, mg_idxopt_t *opt) { int flag = asm_opt.flag; asm_opt.flag |= HA_F_NO_HPC; ha_flt_tab = ha_ft_ug_gen(&asm_opt, &(ug->u), opt->is_HPC, opt->k, opt->w, 1, opt->hap_n*5); ha_idx = ha_pt_ug_gen(&asm_opt, ha_flt_tab, &(ug->u), opt->is_HPC, opt->k, opt->w, 1); asm_opt.flag = flag; fprintf(stderr, "[M::%s] Index has been built.\n", __func__); } void uidx_destory() { ha_ft_destroy(ha_flt_tab); ha_pt_destroy(ha_idx); ha_flt_tab = NULL; ha_idx = NULL; } void mg_gres_a_des(mg_gres_a *p) { uint64_t i = 0; for (i = 0; i < p->n; i++){ free(p->a[i].lc); free(p->a[i].gc); } free(p->a); } ///only use non-repetitive minimizers static mg_match_t *collect_matches(void *km, int *_n_m, int max_occ, const void *ha_flt_tab, const ha_pt_t *ha_idx, int check_unique, const ha_mzl_v *mv, int64_t *n_a, int *rep_len, int *n_mini_pos, int32_t **mini_pos) { int rep_st = 0, rep_en = 0, n_m, tn, tw; size_t i; mg_match_t *m; *n_mini_pos = 0; KMALLOC(km, *mini_pos, mv->n);///mv->n how many minimizers in query m = (mg_match_t*)kmalloc(km, mv->n * sizeof(mg_match_t)); for (i = 0, n_m = 0, *rep_len = 0, *n_a = 0; i < mv->n; ++i) { const ha_idxposl_t *cr; ha_mzl_t *z = &mv->a[i]; cr = ha_ptl_get(ha_idx, z->x, &tn); tw = ha_ft_cnt(ha_flt_tab, z->x); if ((tw > max_occ) || (check_unique && tw != 1)) { ///the frequency of repetitive regions; ignore those minimizers int en = z->pos + 1, st = en - z->span;//[st, en) if (st > rep_en) { ///just record the length of repetive regions *rep_len += rep_en - rep_st; rep_st = st, rep_en = en; } else rep_en = en; } else { mg_match_t *q = &m[n_m++]; q->q_pos = z->pos, q->q_span = z->span, q->rev = z->rev, q->cr = cr, q->n = tn, q->qid = 0; q->is_tandem = 0, q->weight = 255; if(check_unique && tw != 1) q->is_tandem = 1, q->weight = 1; *n_a += q->n;///how many candidates (*mini_pos)[(*n_mini_pos)++] = z->pos;///minimizer offset in query } } *rep_len += rep_en - rep_st; ///the length of repetitive regions *_n_m = n_m; return m; } mg128_t *collect_seed_hits(void *km, const mg_idxopt_t *opt, int max_occ, const void *ha_flt_tab, const ha_pt_t *ha_idx, const ma_ug_t *ug, const ha_mzl_v *mv, int64_t *n_a, int *rep_len, int *n_mini_pos, int32_t **mini_pos) { int i, n_m; mg128_t *a = NULL; mg_match_t *m = collect_matches(km, &n_m, max_occ, ha_flt_tab, ha_idx, 1, mv, n_a, rep_len, n_mini_pos, mini_pos); a = (mg128_t*)kmalloc(km, *n_a * sizeof(mg128_t));///n_a: how many available candidates in total for (i = 0, *n_a = 0; i < n_m; ++i) {///n_m: how many available seeds, instead of candidates mg_match_t *q = &m[i]; const ha_idxposl_t *r = q->cr; uint32_t k; for (k = 0; k < q->n; ++k) {///q->n: number of candidates belonging to seed m[i] mg128_t *p; p = &a[(*n_a)++];///pick up a slot for one candidate if (r[k].rev == q->rev) // forward strand p->x = (uint64_t)(r[k].rid)<<33|r[k].pos; ///reference: rid(31)|rev(1)|pos(32) else // reverse strand p->x = (uint64_t)(r[k].rid)<<33 | 1ULL<<32 | (ug->g->seq[r[k].rid].len - (r[k].pos + 1 - r[k].span) - 1); p->y = (uint64_t)q->q_span << 32 | q->q_pos; p->y |= (uint64_t)q->qid << MG_SEED_SEG_SHIFT; if (q->is_tandem) p->y |= MG_SEED_TANDEM; p->y |= (uint64_t)q->weight << MG_SEED_WT_SHIFT; ///p->y: weight(8)seg_id(8)flag(8)span(8)pos(32) ///p->x: rid(31)rev(1)pos(33); keep reference } } kfree(km, m); radix_sort_128x(a, a + (*n_a)); return a; } ///r is 1000 in default ///remove isolated hits, whic are not close enough to others int64_t flt_anchors(int64_t n_a, mg128_t *a, int32_t r) { int64_t i, j; for (i = 0; i < n_a; ++i) { for (j = i - 1; j >= 0; --j) { /** * a is sorted by x * a[].x: ref_id(31)rev(1)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) **/ int32_t dq; int64_t dr = a[i].x - a[j].x;///a is sorted by x if (dr > r) break;///if two candidates coming from differnt unitigs, dr would be extremly large dq = (int32_t)a[i].y - (int32_t)a[j].y; if (dq > r || dq < 0) continue; a[j].y |= MG_SEED_KEPT; a[i].y |= MG_SEED_KEPT; break; } } for (i = n_a - 1; i >= 0; --i) { if (a[i].y & MG_SEED_KEPT) continue; for (j = i + 1; j < n_a; ++j) { int32_t dq; int64_t dr = a[j].x - a[i].x; if (dr > r) break; dq = (int32_t)a[j].y - (int32_t)a[i].y; if (dq > r || dq < 0) continue; a[j].y |= MG_SEED_KEPT; a[i].y |= MG_SEED_KEPT; break; } } for (i = j = 0; i < n_a; ++i) if (a[i].y & MG_SEED_KEPT) a[j++] = a[i]; return j; } static inline float mg_log2(float x) // NB: this doesn't work when x<2 { union { float f; uint32_t i; } z = { x }; float log_2 = ((z.i >> 23) & 255) - 128; z.i &= ~(255 << 23); z.i += 127 << 23; log_2 += (-0.34484843f * z.f + 2.02466578f) * z.f - 0.67487759f; return log_2; } inline int32_t normal_sc(uint64_t w, int32_t sc) { if(w < 255){ int32_t tmp = (int)(0.00392156862745098 * w * sc); // 0.00392... = 1/255 sc = tmp > 1? tmp : 1; } return sc; } // ai[].x: ref_id(31)rev(1)r_pos(32) // ai[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) // comput_sc(&a[i], &a[j], max_dist_x, max_dist_y, bw, chn_pen_gap, chn_pen_skip, is_cdna, n_segs); static inline int32_t comput_sc(const mg128_t *ai, const mg128_t *aj, int32_t max_dist_x, int32_t max_dist_y, int32_t bw, float chn_pen_gap) { int32_t dq = (int32_t)ai->y - (int32_t)aj->y, dr = (int32_t)ai->x - (int32_t)aj->x, dd, dg, q_span, sc; ///ai and aj has already been sorted by x ///which means ai->x >= aj->x if (dq <= 0 || dq > max_dist_x) return INT32_MIN; if (dr <= 0 || dr > max_dist_y) return INT32_MIN; dd = dr > dq? dr - dq : dq - dr; ///indel, dd is always >= 0 if (dd > bw) return INT32_MIN; dg = dr < dq? dr : dq;///MIN(dr, dq) q_span = aj->y>>32&0xff;///query span; should be ai->y>>32&0xff, is it a bug? sc = normal_sc(aj->y>>MG_SEED_WT_SHIFT, (q_span q_span: there are some bases that are not covered between ai and aj ///it is if (dd || dg > q_span) in minigraph if (dd) { float lin_pen, log_pen; lin_pen = chn_pen_gap * (float)dd; log_pen = dd >= 2? mg_log2(dd) : 0.0f; // mg_log2() only works for dd>=2 sc -= (int)(lin_pen + log_pen); } return sc; } ///p[]: id of last ///f[]: the score ending at i, not always the peak ///v[]: keeps the peak score up to i; ///t[]: used for buffer ///min_cnt = 2; min_sc = 30; extra_u = 0 ///u = mg_chain_backtrack(n, f, p, v, t, min_cnt, min_sc, 0, &n_u, &n_v); uint64_t *mg_chain_backtrack(void *km, int64_t n, const int32_t *f, const int64_t *p, int32_t *v, int32_t *t, int32_t min_cnt, int32_t min_sc, int32_t extra_u, int32_t *n_u_, int32_t *n_v_) { mg128_t *z; uint64_t *u; int64_t i, k, n_z, n_v; int32_t n_u; // v[] keeps the peak score up to i; f[] is the score ending at i, not always the peak *n_u_ = *n_v_ = 0; for (i = 0, n_z = 0; i < n; ++i) // precompute n_z if (f[i] >= min_sc) ++n_z; if (n_z == 0) return 0; KMALLOC(km, z, n_z); for (i = 0, k = 0; i < n; ++i) // populate z[] if (f[i] >= min_sc) z[k].x = f[i], z[k++].y = i; radix_sort_128x(z, z + n_z);///sort by score memset(t, 0, n * 4);///t is a buffer ///from the largest to the smallest for (k = n_z - 1, n_v = n_u = 0; k >= 0; --k) { // precompute n_u int64_t n_v0 = n_v; int32_t sc; ///note t[i] == 0 is not used to find local alignment ///say if we have already found a long chain, then the secondary might be able to merged to the long chain ///t[i] == 0 is used to find those chains for (i = z[k].y; i >= 0 && t[i] == 0; i = p[i]) ++n_v, t[i] = 1; sc = i < 0? z[k].x : (int32_t)z[k].x - f[i]; if (sc >= min_sc && n_v > n_v0 && n_v - n_v0 >= min_cnt) ++n_u;///how many chains, including primary chains and non-primary chains else n_v = n_v0; } KMALLOC(km, u, n_u + extra_u); memset(t, 0, n * 4); for (k = n_z - 1, n_v = n_u = 0; k >= 0; --k) { // populate u[] int64_t n_v0 = n_v; int32_t sc; for (i = z[k].y; i >= 0 && t[i] == 0; i = p[i]) v[n_v++] = i, t[i] = 1; sc = i < 0? z[k].x : (int32_t)z[k].x - f[i]; if (sc >= min_sc && n_v > n_v0 && n_v - n_v0 >= min_cnt) u[n_u++] = (uint64_t)sc << 32 | (n_v - n_v0); else n_v = n_v0; } kfree(km, z); assert(n_v < INT32_MAX); *n_u_ = n_u, *n_v_ = n_v; return u; } //u[]: sc|occ of chains //v[]: idx of each element static mg128_t *compact_a(void *km, int32_t n_u, uint64_t *u, int32_t n_v, int32_t *v, mg128_t *a) { mg128_t *b, *w; uint64_t *u2; int64_t i, j, k; // write the result to b[] KMALLOC(km, b, n_v); for (i = 0, k = 0; i < n_u; ++i) { int32_t k0 = k, ni = (int32_t)u[i]; for (j = 0; j < ni; ++j) b[k++] = a[v[k0 + (ni - j - 1)]];///write all elements of a chain together } kfree(km, v); // sort u[] and a[] by the target position, such that adjacent chains may be joined KMALLOC(km, w, n_u); for (i = k = 0; i < n_u; ++i) {///n_u: how many chains ///x: ref_id(31)rev(1)r_pos(32) w[i].x = b[k].x, w[i].y = (uint64_t)k<<32|i; k += (int32_t)u[i]; } radix_sort_128x(w, w + n_u);///sort by ref_id(31)rev(1)r_pos(32); r_pos is the start pos of chain KMALLOC(km, u2, n_u); for (i = k = 0; i < n_u; ++i) {///note merge chain; just place close chains together ///j is chain id; n is how many elements in j-th chain int32_t j = (int32_t)w[i].y, n = (int32_t)u[j]; u2[i] = u[j]; memcpy(&a[k], &b[w[i].y>>32], n * sizeof(mg128_t)); k += n; } memcpy(u, u2, n_u * 8); memcpy(b, a, k * sizeof(mg128_t)); // write _a_ to _b_ and deallocate _a_ because _a_ is oversized, sometimes a lot kfree(km, a); kfree(km, w); kfree(km, u2); return b; } /* Input: * a[].x: ref_id(31)rev(1)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) * n: length of a[] * Output: * n_u: #chains * u[]: score<<32 | #anchors (sum of lower 32 bits of u[] is the returned length of a[]) * input a[] is deallocated on return */ ///is_cdna is is_splice mg128_t *mg_lchain_dp(int max_dist_x, int max_dist_y, int bw, int max_skip, int max_iter, int min_cnt, int min_sc, float chn_pen_gap, int64_t n, mg128_t *a, int *n_u_, uint64_t **_u, void *km) { // TODO: make sure this works when n has more than 32 bits int32_t *f, *t, *v, n_u, n_v; int64_t *p, i, j, max_ii, st = 0; uint64_t *u; if (_u) *_u = 0, *n_u_ = 0; if (n == 0 || a == 0) return 0; KMALLOC(km, p, n);///id of last cell KMALLOC(km, f, n);///f[] is the score ending at i, not always the peak KMALLOC(km, v, n);///v[] keeps the peak score up to i; KCALLOC(km, t, n);///t doesn't matter too much; it is mainly used to accelrate the iteration // a[].x: ref_id(31)rev(1)r_pos(32) // a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) // fill the score and backtrack arrays for (i = st = 0, max_ii = -1; i < n; ++i) { int64_t max_j = -1, end_j; ///max_f -> score of minimizer int32_t max_f = normal_sc(a[i].y>>MG_SEED_WT_SHIFT, a[i].y>>32&0xff), n_skip = 0; ///until we are at the same rid, same direction, and the coordinates are close enough while (st < i && (a[i].x>>32 != a[st].x>>32 || a[i].x > a[st].x + max_dist_x)) ++st; ///max_iter = 10000 in default, which means dp can go back to up to 10000 cells if (i - st > max_iter) st = i - max_iter; for (j = i - 1; j >= st; --j) { int32_t sc; sc = comput_sc(&a[i], &a[j], max_dist_x, max_dist_y, bw, chn_pen_gap); if (sc == INT32_MIN) continue; sc += f[j]; if (sc > max_f) { max_f = sc, max_j = j; if (n_skip > 0) --n_skip; } else if (t[j] == (int32_t)i) {///note we scan j backwards; we don't need to update t[] for each i if (++n_skip > max_skip) break; } if (p[j] >= 0) t[p[j]] = i;//p[]: prefix idx; means there is a chain longer than 2 } end_j = j;///end_j might be > 0; just the end idx of backwards ///if not close enough, select a new max ///max_ii is just used to rescue best-score in case best-score appears before end_j if (max_ii < 0 || (int64_t)(a[i].x - a[max_ii].x) > (int64_t)max_dist_x) {///select a new max int32_t max = INT32_MIN; max_ii = -1; for (j = i - 1; j >= st; --j) if (max < f[j]) max = f[j], max_ii = j; } ///note: it will happen when `max_ii` < `end_j`; ///iteration is terminated at `end_j` mostly because of `max_skip` and `max_iter` ///max_ii is just used to rescue best-score in case best-score appears before end_j if (max_ii >= 0 && max_ii < end_j) { int32_t tmp; tmp = comput_sc(&a[i], &a[max_ii], max_dist_x, max_dist_y, bw, chn_pen_gap); if (tmp != INT32_MIN && max_f < tmp + f[max_ii]) max_f = tmp + f[max_ii], max_j = max_ii; } // v[] keeps the peak score up to i (as score might decerase); f[] is the score ending at i, not always the peak f[i] = max_f, p[i] = max_j;//p[]: prefix idx v[i] = max_j >= 0 && v[max_j] > max_f? v[max_j] : max_f; if (max_ii < 0 || ((int64_t)(a[i].x - a[max_ii].x) <= (int64_t)max_dist_x && f[max_ii] < f[i])) max_ii = i; } ///after mg_chain_backtrack, the results are saved in u and v; u = mg_chain_backtrack(km, n, f, p, v, t, min_cnt, min_sc, 0, &n_u, &n_v); *n_u_ = n_u, *_u = u; // NB: note that u[] may not be sorted by score here kfree(km, p); kfree(km, f); kfree(km, t); if (n_u == 0) { kfree(km, a); kfree(km, v); return 0; } //u[]: sc|occ of chains; chain is mostly sorted by the score; at least the first chain has the largest score //v[]: idx of each element return compact_a(km, n_u, u, n_v, v, a); } void extend_coordinates(mg_lchain_t *ri, int64_t qlen, int64_t rlen) { int64_t qs, qe, rs, re, qtail, rtail; qs = ri->qs; qe = ri->qe - 1; rs = ri->rs; re = ri->re - 1; if(ri->v&1) { rs = rlen - ri->re; re = rlen - ri->rs - 1; } if(qs <= rs) { rs -= qs; qs = 0; } else { qs -= rs; rs = 0; } qtail = qlen - qe - 1; rtail = rlen - re - 1; if(qtail <= rtail) { qe = qlen - 1; re += qtail; } else { re = rlen - 1; qe += rtail; } ri->qs = qs; ri->qe = qe + 1; ri->rs = rs; ri->re = re + 1; if(ri->v&1) { ri->rs = rlen - re - 1; ri->re = rlen - rs; } } ///qlen: query length ///u[]: sc|occ of chains ///a[]: candidate list mg_lchain_t *mg_lchain_gen(void *km, int qlen, int n_u, uint64_t *u, mg128_t *a, const ma_ug_t *ug) { mg128_t *z; mg_lchain_t *r; int i, k; if (n_u == 0) return 0; KCALLOC(km, r, n_u); KMALLOC(km, z, n_u); // u[] is sorted by query position for (i = k = 0; i < n_u; ++i) { /** * a[].x: ref_id(31)rev(1)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) **/ ///u[]: sc(32)occ(32) int32_t qs = (int32_t)a[k].y + 1 - (a[k].y>>32 & 0xff); z[i].x = (uint64_t)qs << 32 | u[i] >> 32; z[i].y = (uint64_t)k << 32 | (int32_t)u[i]; k += (int32_t)u[i]; } radix_sort_128x(z, z + n_u);//sort by qs|sc // populate r[] for (i = 0; i < n_u; ++i) { mg_lchain_t *ri = &r[i]; /** * z[].x: query start pos| chain score * z[].y: idx in a[] | chain occ * a[].x: ref_id(31)rev(1)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) * **/ int32_t k = z[i].y >> 32, q_span = a[k].y >> 32 & 0xff; ri->off = k; ri->cnt = (int32_t)z[i].y; ri->score = (uint32_t)z[i].x; ri->v = a[k].x >> 32;///ref_id|rev ri->rs = (int32_t)a[k].x + 1 > q_span? (int32_t)a[k].x + 1 - q_span : 0; // for HPC k-mer ri->qs = z[i].x >> 32; ri->re = (int32_t)a[k + ri->cnt - 1].x + 1; ri->qe = (int32_t)a[k + ri->cnt - 1].y + 1; // fprintf(stderr, "+0+\tA\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%d\tts:%u\tte:%u\ttl:%u\n", // (ri->v>>1)+1, "lc"[ug->u.a[ri->v>>1].circ], "+-"[ri->v&1], ri->qs, ri->qe, qlen, ri->rs, ri->re, ug->u.a[ri->v>>1].len); // extend_coordinates(ri, qlen, ug->u.a[ri->v>>1].len); // fprintf(stderr, "-0-\tA\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%d\tts:%u\tte:%u\ttl:%u\n", // (ri->v>>1)+1, "lc"[ug->u.a[ri->v>>1].circ], "+-"[ri->v&1], ri->qs, ri->qe, qlen, ri->rs, ri->re, ug->u.a[ri->v>>1].len); } kfree(km, z); return r; } static int32_t get_mini_idx(const mg128_t *a, int32_t n, const int32_t *mini_pos) { int32_t x, L = 0, R = n - 1; x = (int32_t)a->y; while (L <= R) { // binary search int32_t m = ((uint64_t)L + R) >> 1; int32_t y = mini_pos[m]; if (y < x) L = m + 1; else if (y > x) R = m - 1; else return m; } return -1; } /* Before: * a[].x: ref_id(31)rev(1)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) * After: * a[].x: idx_in_minimizer_arr(32)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) */ void mg_update_anchors(int32_t n_a, mg128_t *a, int32_t n, const int32_t *mini_pos) { int32_t st, j, k; if (n_a <= 0) return; st = get_mini_idx(&a[0], n, mini_pos); assert(st >= 0); for (k = 0, j = st; j < n && k < n_a; ++j) if ((int32_t)a[k].y == mini_pos[j]) a[k].x = (uint64_t)j << 32 | (a[k].x & 0xffffffffU), ++k; assert(k == n_a); } static int32_t find_max(int32_t n, const gc_frag_t *gf, uint32_t x) { int32_t s = 0, e = n; if (n == 0) return -1; if (gf[n-1].srt < x) return n - 1; if (gf[0].srt >= x) return -1; while (e > s) { // TODO: finish this block int32_t m = s + (e - s) / 2; if (gf[m].srt >= x) e = m; else s = m + 1; } assert(s == e); return s; } ///target_dist should like the overlap length in string graph ///it should be used to evaluate if the identified path is close to real path/alignment static int32_t mg_target_dist(const asg_t *g, const mg_lchain_t *l0, const mg_lchain_t *l1) { /** case 1: l0->qs************l0->qe l1->qs************l1->qe case 2: l0->qs************l0->qe l1->qs************l1->qe *****l0->rs************l0->re***** ****l1->rs************l1->re** * **/ ///min_dist = l1->rs + (g->seg[l0->v>>1].len - l0->re); // below equals (l1->qs - l0->qe) - min_dist + g->seg[l1->v>>1].len; see mg_gchain1_dp() for the calculation of min_dist //(l1->qs - l0->qe) is the gap in query, min_dist is the gap in reference return (l1->qs - l0->qe) - (g->seq[l0->v>>1].len - l0->re) + (g->seq[l1->v>>1].len - l1->rs); // when l0->v == l1->v, the above becomes (l1->qs - l0->qe) - (l1->rs - l0->re), which is what we want } static inline sp_node_t *gen_sp_node(void *km, uint32_t v, int32_t d, int32_t id) { sp_node_t *p; KMALLOC(km, p, 1); p->v = v, p->di = (uint64_t)d<<32 | id, p->pre = -1, p->is_0 = 1; return p; } ///max_dist is like the overlap length in string graph ///the end position of qs is li->qs; dst[]->->qlen indicate the region that need to be checked in bases mg_pathv_t *mg_shortest_k(void *km0, const asg_t *g, uint32_t src, int32_t n_dst, mg_path_dst_t *dst, int32_t max_dist, int32_t max_k, /** //pathint32_t ql, const char *qs, int is_rev, **/int32_t *n_pathv) { sp_node_t *p, *root = 0, **out; sp_topk_t *q; khash_t(sp) *h; khash_t(sp2) *h2; void *km; khint_t k; int absent; int32_t i, j, n_done, n_found; uint32_t id, n_out, m_out; int8_t *dst_done; mg_pathv_t *ret = 0; uint64_t *dst_group; /** //path int32_t n_seeds = 0; uint64_t *seeds = 0; void *h_seeds = 0; mg128_v mini = {0,0,0}; **/ if (n_pathv) *n_pathv = 0;///for us, n_pathv = NULL if (n_dst <= 0) return 0;///n_dst: how many candidate nodes for (i = 0; i < n_dst; ++i) { // initialize mg_path_dst_t *t = &dst[i]; ///if src and dest are at the same ref id, there are already one path if (t->inner)///if two chains are at the same ref id t->dist = 0, t->n_path = 1, t->path_end = -1; else t->dist = -1, t->n_path = 0, t->path_end = -1; } if (max_k > MG_MAX_SHORT_K) max_k = MG_MAX_SHORT_K; km = km_init2(km0, 0x4000); /** //path ///for the first time, we just check th reachability without sequence (qs); ///but for the second round, we need to check sequence ///qs is the sequence between two minimizers if (ql > 0 && qs) { // build the seed hash table for the query mg_sketch(km, qs, ql, MG_SHORT_KW, MG_SHORT_KK, 0, &mini); // mini->a[].x = hash_key<<8 | kmerSpan // mini->a[].y = rid<<32 | lastPos<<1 | strand if (is_rev)///is_rev = 1; for (i = 0; i < mini.n; ++i)///reverse qs[0, ql) to qs(ql, 0] mini.a[i].y = (ql - (((int32_t)mini.a[i].y>>1) + 1 - MG_SHORT_KK) - 1) << 1 | ((mini.a[i].y&1)^1); ///h_seeds is the ordinary hash index h_seeds = mg_idx_a2h(km, mini.n, mini.a, 0, &seeds, &n_seeds); ///h_seeds+seeds+n_seeds ----> hash index of qs[0, ql) } **/ ///dst is how many candidates KCALLOC(km, dst_done, n_dst); KMALLOC(km, dst_group, n_dst); // multiple dst[] may have the same dst[].v. We need to group them first. // in other words, one ref id may have multiple dst alignment chains for (i = 0; i < n_dst; ++i) dst_group[i] = (uint64_t)dst[i].v<<32 | i; radix_sort_gfa64(dst_group, dst_group + n_dst); h2 = kh_init2(sp2, km); // (h2+dst_group) keeps all destinations from the same ref id kh_resize(sp2, h2, n_dst * 2); ///please note that one contig in ref may have multiple alignment chains ///so h2 is a index that helps us to query it ///key(h2) = ref id; value(h2) = start_idx | occ for (i = 1, j = 0; i <= n_dst; ++i) { if (i == n_dst || dst_group[i]>>32 != dst_group[j]>>32) { k = kh_put(sp2, h2, dst_group[j]>>32, &absent); kh_val(h2, k) = (uint64_t)j << 32 | (i - j); assert(absent); j = i; } } h = kh_init2(sp, km); // h keeps visited vertices; path to each visited vertice kh_resize(sp, h, 16); m_out = 16, n_out = 0;///16 is just the initial size KMALLOC(km, out, m_out); /** typedef struct { int32_t k, mlen;//k: number of walks from src to this node int32_t qs, qe; sp_node_t *p[MG_MAX_SHORT_K]; // this forms a max-heap; all path } sp_topk_t; **/ id = 0; p = gen_sp_node(km, src, 0, id++);///just malloc a node for src; the distance is 0 p->hash = __ac_Wang_hash(src);///hash is path hash, instead of node hash kavl_insert(sp, &root, p, 0);///should be avl tree ///each src corresponds to one node in the hash table , but corresponds to node in the AVL tree k = kh_put(sp, h, src, &absent);///here is a hash table q = &kh_val(h, k); ///for normal graph traversal, one node just has one parental node; here each node has at most 16 parental nodes q->k = 1, q->p[0] = p, q->mlen = 0, q->qs = q->qe = -1; n_done = 0; ///the key of avl tree: #define sp_node_cmp(a, b) (((a)->di > (b)->di) - ((a)->di < (b)->di)) ///the higher bits of (*)->di is distance to src node ///so the key of avl tree is distance ///in avl tree , one node might be saved multipe times while (kavl_size(head, root) > 0) {///thr first root is src int32_t i, nv; asg_arc_t *av; sp_node_t *r; ///note that one (sp_node_t->v) might be visited multiple times if there are circles ///so there might be multipe nodes with the same (sp_node_t->v) ///delete the first node r = kavl_erase_first(sp, &root); // take out the closest vertex in the heap (as a binary tree) //fprintf(stderr, "XX\t%d\t%d\t%d\t%c%s[%d]\t%d\n", n_out, kavl_size(head, root), n_finished, "><"[(r->v&1)^1], g->seg[r->v>>1].name, r->v, (int32_t)(r->di>>32)); if (n_out == m_out) KEXPAND(km, out, m_out); ///higher 32 bits might be the distance to root node // lower 32 bits now for position in the out[] array r->di = r->di>>32<<32 | n_out; ///n_out is just the id in out ///so one node id in graph might be saved multiple times in avl tree and out[] out[n_out++] = r;///out[0] = src ///r->v is the dst vertex id ///sometimes k==kh_end(h2). Some nodes are found by graph travesal but not in linear chain alignment k = kh_get(sp2, h2, r->v); // we have reached one dst vertex // note that one dst vertex may have multipe alignment chains // we can visit some nodes in graph which are not reachable during chaining // h2 is used to determine if one node is reachable or not if (k != kh_end(h2)) { ///node r->v might be visited multiple times int32_t j, dist = r->di>>32, off = kh_val(h2, k) >> 32, cnt = (int32_t)kh_val(h2, k); //src can reach ref id r->v; there might be not only one alignment chain in r->v //so we need to scan all of them for (j = 0; j < cnt; ++j) { mg_path_dst_t *t = &dst[(int32_t)dst_group[off + j]];///t is a linear alignment at r->v int32_t done = 0; ///the src and dest are at the same ref id, say we directly find the shortest path if (t->inner) {//usually the first node, which is same to src done = 1; } else { int32_t mlen = 0, copy = 0; ///in the first round, we just check reachability without sequence ///so h_seeds = NULL; we can assume mlen = 0 /** //path mlen = h_seeds? path_mlen(out, n_out - 1, h, t->qlen) : 0; **/ //if (mg_dbg_flag & MG_DBG_GC1) fprintf(stderr, " src=%c%s[%d],qlen=%d\tdst=%c%s[%d]\ttarget_distx=%d,target_hash=%x\tdistx=%d,mlen=%d,hash=%x\n", "><"[src&1], g->seg[src>>1].name, src, ql, "><"[t->v&1], g->seg[t->v>>1].name, t->v, t->target_dist - g->seg[src>>1].len, t->target_hash, dist - g->seg[src>>1].len, mlen, r->hash); // note: t indicates a linear alignmnet, instead of a node in graph ///target_dist should be the distance on query if (t->n_path == 0) { // means this alignment has never been visited before; keep the shortest path anyway copy = 1; // we have a target distance; choose the closest; // there is already several paths reaching the linear alignment } else if (t->target_dist >= 0) { // we found the target path; hash is the path hash including multiple nodes, instead of node hash if (dist == t->target_dist && t->check_hash && r->hash == t->target_hash) { copy = 1, done = 1; } else { int32_t d0 = t->dist, d1 = dist; d0 = d0 > t->target_dist? d0 - t->target_dist : t->target_dist - d0; d1 = d1 > t->target_dist? d1 - t->target_dist : t->target_dist - d1; ///if the new distance (d1) is smaller than the old distance (d0), update the results ///the length of new path should be closer to t->target_dist if (d1 - mlen/2 < d0 - t->mlen/2) copy = 1; } } if (copy) { t->path_end = n_out - 1, t->dist = dist, t->hash = r->hash, t->mlen = mlen, t->is_0 = r->is_0; if (t->target_dist >= 0) { ///src is from li from li to lj, so the dis is generally increased; dijkstra algorithm ///target_dist should be the distance on query if (dist == t->target_dist && t->check_hash && r->hash == t->target_hash) done = 1; else if ((dist > t->target_dist + MG_SHORT_K_EXT) && (dist > (t->target_dist>>4))) done = 1; } } ++t->n_path;///we found a path to the alignment t if (t->n_path >= max_k) done = 1; } if (dst_done[off + j] == 0 && done) dst_done[off + j] = 1, ++n_done; } ///if all alignments have been settle down ///pre-end; accelerate the loop if (n_done == n_dst) break; } ///below is used to push new nodes to avl tree for iteration nv = asg_arc_n(g, r->v); av = asg_arc_a(g, r->v); for (i = 0; i < nv; ++i) { // visit all neighbors asg_arc_t *ai = &av[i]; ///v_lv is the (dest_length - overlap_length); it is a normal path length in string graph ///ai->v_lv is the path length from r->v to ai->w ///(r->di>>32) int32_t d = (r->di>>32) + (uint32_t)ai->ul; if (d > max_dist) continue; // don't probe vertices too far away // h keeps visited vertices; path to each visited vertice ///ai->w is the dest ref id; we insert a new ref id, instead of an alignment chain k = kh_put(sp, h, ai->v, &absent);///one node might be visited multiple times q = &kh_val(h, k); if (absent) { // a new vertex visited ///q->k: number of walks from src to ai->w q->k = 0, q->qs = q->qe = -1; q->mlen = 0; ///h_seeds = NULL; so q->mlen = 0 /** //path q->mlen = h_seeds && d + gfa_arc_lw(g, *ai) <= max_dist? node_mlen(km, g, ai->w, &mini, h_seeds, n_seeds, seeds, &q->qs, &q->qe) : 0; **/ //if (ql && qs) fprintf(stderr, "ql=%d,src=%d\tv=%c%s[%d],n_seeds=%d,mlen=%d\n", ql, src, "><"[ai->w&1], g->seg[ai->w>>1].name, ai->w, n_seeds, q->mlen); } ///if there are less than walks from src to ai->w, directly add ///if there are more, keep the smallest walks if (q->k < max_k) { // enough room: add to the heap p = gen_sp_node(km, ai->v, d, id++); p->pre = n_out - 1;///the parent node of this one p->hash = r->hash + __ac_Wang_hash(ai->v); p->is_0 = r->is_0; /** //path if (ai->rank > 0) p->is_0 = 0; **/ kavl_insert(sp, &root, p, 0); q->p[q->k++] = p; ks_heapup_sp(q->k, q->p);///adjust heap by distance } else if ((int32_t)(q->p[0]->di>>32) > d) { // shorter than the longest path so far: replace the longest p = kavl_erase(sp, &root, q->p[0], 0); if (p) { p->di = (uint64_t)d<<32 | (id++); p->pre = n_out - 1; p->hash = r->hash + __ac_Wang_hash(ai->v); p->is_0 = r->is_0; /** //path if (ai->rank > 0) p->is_0 = 0; **/ kavl_insert(sp, &root, p, 0); ks_heapdown_sp(0, q->k, q->p); } else { fprintf(stderr, "Warning: logical bug in gfa_shortest_k(): q->k=%d,q->p[0]->{d,i}={%d,%d},d=%d,src=%u,max_dist=%d,n_dst=%d\n", q->k, (int32_t)(q->p[0]->di>>32), (int32_t)q->p[0]->di, d, src, max_dist, n_dst); km_destroy(km); return 0; } } // else: the path is longer than all the existing paths ended at ai->w } } kfree(km, dst_group); kfree(km, dst_done); kh_destroy(sp, h); /** //path mg_idx_hfree(h_seeds); kfree(km, seeds); kfree(km, mini.a); **/ // NB: AVL nodes are not deallocated. When km==0, they are memory leaks. for (i = 0, n_found = 0; i < n_dst; ++i) if (dst[i].n_path > 0) ++n_found;///n_path might be larger than 16 ///we can assume n_pathv = NULL for now if (n_found > 0 && n_pathv) { // then generate the backtrack array int32_t n, *trans; ///n_out: how many times that nodes in graph have been visited ///note one node might be visited multiples times KCALLOC(km, trans, n_out); // used to squeeze unused elements in out[] ///n_dst: number of alignment chains for (i = 0; i < n_dst; ++i) { // mark dst vertices with a target distance mg_path_dst_t *t = &dst[i]; if (t->n_path > 0 && t->target_dist >= 0 && t->path_end >= 0) trans[(int32_t)out[t->path_end]->di] = 1;///(int32_t)out[]->di: traverse track corresponds to the alignment chain dst[] } for (i = 0; (uint32_t)i < n_out; ++i) { // mark dst vertices without a target distance k = kh_get(sp2, h2, out[i]->v); if (k != kh_end(h2)) { // TODO: check if this is correct! int32_t off = kh_val(h2, k)>>32, cnt = (int32_t)kh_val(h2, k); for (j = off; j < off + cnt; ++j) if (dst[j].target_dist < 0) trans[i] = 1; } } for (i = n_out - 1; i >= 0; --i) // mark all predecessors if (trans[i] && out[i]->pre >= 0) trans[out[i]->pre] = 1; for (i = n = 0; (uint32_t)i < n_out; ++i) // generate coordinate translations if (trans[i]) trans[i] = n++; else trans[i] = -1; *n_pathv = n; KMALLOC(km0, ret, n); for (i = 0; (uint32_t)i < n_out; ++i) { // generate the backtrack array mg_pathv_t *p; if (trans[i] < 0) continue; p = &ret[trans[i]]; p->v = out[i]->v, p->d = out[i]->di >> 32; p->pre = out[i]->pre < 0? out[i]->pre : trans[out[i]->pre]; } for (i = 0; i < n_dst; ++i) // translate "path_end" if (dst[i].path_end >= 0) dst[i].path_end = trans[dst[i].path_end]; } km_destroy(km); return ret; } static inline int32_t cal_sc(const mg_path_dst_t *dj, const mg_lchain_t *li, const mg_lchain_t *lc, const mg128_t *an, const gc_frag_t *a, const int32_t *f, int bw, int ref_bonus, float chn_pen_gap) { const mg_lchain_t *lj; int32_t gap, sc; float lin_pen, log_pen; if (dj->n_path == 0) return INT32_MIN; gap = dj->dist - dj->target_dist; lj = &lc[a[dj->meta].i]; if (gap < 0) gap = -gap; if (gap > bw) return INT32_MIN; if (lj->qe <= li->qs) sc = li->score; else sc = (int32_t)((double)(li->qe - lj->qe) / (li->qe - li->qs) * li->score + .499); // dealing with overlap on query //sc += dj->mlen; // TODO: is this line the right thing to do? if (dj->is_0) sc += ref_bonus; lin_pen = chn_pen_gap * (float)gap; log_pen = gap >= 2? mg_log2(gap) : 0.0f; sc -= (int32_t)(lin_pen + log_pen); sc += f[dj->meta]; return sc; } void transfor_icoord(const int64_t iqs, const int64_t iqe, const int64_t irs, const int64_t ire, const uint8_t rev, const int64_t qlen, const int64_t rlen, int32_t *r_qs, int32_t *r_qe, int32_t *r_rs, int32_t *r_re) { int64_t qs, qe, rs, re, qtail, rtail; qs = iqs; qe = iqe - 1; rs = irs; re = ire - 1; if(rev) { rs = rlen - ire; re = rlen - irs - 1; } if(qs <= rs) { rs -= qs; qs = 0; } else { qs -= rs; rs = 0; } qtail = qlen - qe - 1; rtail = rlen - re - 1; if(qtail <= rtail) { qe = qlen - 1; re += qtail; } else { re = rlen - 1; qe += rtail; } if(r_qs) (*r_qs) = qs; if(r_qe) (*r_qe) = qe + 1; if(r_rs) (*r_rs) = rs; if(r_re) (*r_re) = re + 1; if(rev) { if(r_rs) (*r_rs) = rlen - re - 1; if(r_re) (*r_re) = rlen - rs; } } void transfor_coord(mg_lchain_t *ri, const int64_t qlen, const int64_t rlen, int32_t *r_qs, int32_t *r_qe, int32_t *r_rs, int32_t *r_re) { int64_t qs, qe, rs, re, qtail, rtail; qs = ri->qs; qe = ri->qe - 1; rs = ri->rs; re = ri->re - 1; if(ri->v&1) { rs = rlen - ri->re; re = rlen - ri->rs - 1; } if(qs <= rs) { rs -= qs; qs = 0; } else { qs -= rs; rs = 0; } qtail = qlen - qe - 1; rtail = rlen - re - 1; if(qtail <= rtail) { qe = qlen - 1; re += qtail; } else { re = rlen - 1; qe += rtail; } if(r_qs) (*r_qs) = qs; if(r_qe) (*r_qe) = qe + 1; if(r_rs) (*r_rs) = rs; if(r_re) (*r_re) = re + 1; if(ri->v&1) { if(r_rs) (*r_rs) = rlen - re - 1; if(r_re) (*r_re) = rlen - rs; } } int64_t get_nn_ov(const uint32_t v, const uint32_t w, const asg_t *g) { uint32_t i; uint32_t nv = asg_arc_n(g, v); asg_arc_t *av = asg_arc_a(g, v), *p = NULL; for (i = 0; i < nv; i++) { if(av[i].del) continue; if(av[i].v == w) { // o -= av[i].ol; p = &(av[i]); break; } } return p?p->ol:0; } int64_t get_lchain_ovlp(mg_lchain_t *lp, mg_lchain_t *la, const asg_t *g, const int64_t qlen, const ma_ug_t *ug) { int64_t o = lp->qe - la->qs, oj; uint32_t v = la->v^1, w = lp->v^1, i; int32_t pqe, aqs; if(o <= 0) return 0; if(v == w) return o; transfor_coord(lp, qlen, ug->u.a[lp->v>>1].len, NULL, &pqe, NULL, NULL); transfor_coord(la, qlen, ug->u.a[la->v>>1].len, &aqs, NULL, NULL, NULL); uint32_t nv = asg_arc_n(g, v); asg_arc_t *av = asg_arc_a(g, v), *p = NULL; for (i = 0; i < nv; i++) { if(av[i].del) continue; if(av[i].v == w) { // o -= av[i].ol; p = &(av[i]); break; } } oj = o; if(p) oj = pqe - aqs - p->ol; if(o > oj) o = oj; if(o < 0) o = 0; return o; } int64_t get_lchain_gap(mg_lchain_t *lp, mg_lchain_t *la, const asg_t *g, const int64_t qlen, const ma_ug_t *ug, int32_t double_ol) { int64_t gg = la->qs - lp->qe, ggj; uint32_t v = la->v^1, w = lp->v^1, i; int32_t aqs, pqe; if(double_ol == 0 && gg >= 0) return gg; if(v == w) return gg; transfor_coord(la, qlen, ug->u.a[la->v>>1].len, &aqs, NULL, NULL, NULL); transfor_coord(lp, qlen, ug->u.a[lp->v>>1].len, NULL, &pqe, NULL, NULL); uint32_t nv = asg_arc_n(g, v); asg_arc_t *av = asg_arc_a(g, v), *p = NULL;; for (i = 0; i < nv; i++) { if(av[i].del) continue; if(av[i].v == w) { // gg += av[i].ol; p = &(av[i]); break; } } ggj = gg; if(p) ggj = aqs - pqe + p->ol + (double_ol?p->ol:0); // if(gg < ggj) gg = ggj; // return gg; return ggj; } int64_t max_ovlp(const asg_t *g, uint32_t v) { uint32_t i, nv = asg_arc_n(g, v), o = 0; asg_arc_t *av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].del) continue; if(o < av[i].ol) o = av[i].ol; } return o; } int64_t max_ovlp_src(const ug_opt_t *uopt, uint32_t v) { ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp, max_hang = uopt->max_hang; uint32_t i, qn, tn, o = 0, x = v>>1; asg_arc_t e; for (i = 0; i < src[x].length; i++) { qn = Get_qn(src[x].buffer[i]); tn = Get_tn(src[x].buffer[i]); if(ma_hit2arc(&(src[x].buffer[i]), Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &e) < 0) { continue; } if((e.ul>>32) != v) continue; if(o < e.ol) o = e.ol; } return o; } int64_t specific_ovlp(const ma_ug_t *ug, const ug_opt_t *uopt, const uint32_t v, const uint32_t w) { if(ug->u.a[v>>1].circ || ug->u.a[w>>1].circ) return 0; uint32_t rv, rw, i; int32_t r; const ma_hit_t_alloc *x = NULL; asg_arc_t t; memset(&t, 0, sizeof(t)); if(v&1) rv = ug->u.a[v>>1].start^1; else rv = ug->u.a[v>>1].end^1; if(w&1) rw = ug->u.a[v>>1].end; else rw = ug->u.a[v>>1].start; x = &(uopt->sources[rv>>1]); for (i = 0; i < x->length; i++) { if(Get_tn(x->buffer[i]) == (rw>>1)) { r = ma_hit2arc(&(x->buffer[i]), uopt->coverage_cut[rv>>1].e - uopt->coverage_cut[rv>>1].s, uopt->coverage_cut[rw>>1].e - uopt->coverage_cut[rw>>1].s, uopt->max_hang, asm_opt.max_hang_rate, uopt->min_ovlp, &t); if(r < 0) return 0; if((t.ul>>32)!=rv || t.v!=rw) return 0; return t.ol; } } return 0; } void extend_lchain(mg_lchain_t *lc, int32_t n_lc, int32_t qlen, const ma_ug_t *ug) { int32_t i; for (i = 0; i < n_lc; ++i) { extend_coordinates(&lc[i], qlen, ug->u.a[lc[i].v>>1].len); } } void compress_lchain(mg_lchain_t *lc, int32_t n_lc, int32_t qlen, const ma_ug_t *ug, const mg128_t *a) { int32_t i, k, q_span; mg_lchain_t *ri = NULL; for (i = 0; i < n_lc; ++i) { ri = &lc[i]; k = ri->off; ri->rs = (int32_t)a[k].x + 1 > q_span? (int32_t)a[k].x + 1 - q_span : 0; // for HPC k-mer ri->qs = (int32_t)a[k].y + 1 - (a[k].y>>32 & 0xff); ri->re = (int32_t)a[k + ri->cnt - 1].x + 1; ri->qe = (int32_t)a[k + ri->cnt - 1].y + 1; } } void print_gchain(gc_frag_t *a, const int64_t *p, mg_lchain_t *lc, const int64_t nlc, const ma_ug_t *ug, int32_t qlen) { int64_t k, i; gc_frag_t *ai = NULL; mg_lchain_t *li = NULL; for (k = 0; k < nlc; k++) { fprintf(stderr, "\n"); for (i = k; i >= 0; i = p[i]) { ai = &a[i]; li = &lc[ai->i]; fprintf(stderr, "*\tXXXXXX\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%d\tts:%u\tte:%u\ttl:%u\n", (li->v>>1)+1, "lc"[ug->u.a[li->v>>1].circ], "+-"[li->v&1], li->qs, li->qe, qlen, li->rs, li->re, ug->u.a[li->v>>1].len); } } } // void extend_graph_coordnates(const ma_ug_t *ug, const ug_opt_t *uopt, const mg_lchain_t *lp, const mg_lchain_t *la, // mg_coor_t *gp, mg_coor_t *ga, int32_t *go, int32_t *gg) // { // int32_t so = specific_ovlp(ug, uopt, lp->v^1, la->v^1); // } int32_t mg_gchain1_dp(void *km, const ma_ug_t *ug, const asg_t *rg, int32_t *n_lc_, mg_lchain_t *lc, int32_t qlen, int32_t max_dist_g, int32_t max_dist_q, int32_t bw, int32_t max_skip, int32_t ref_bonus, float chn_pen_gap, float mask_level, int32_t max_gc_seq_ext, const ug_opt_t *uopt, const mg128_t *an, uint64_t **u_) { int32_t i, j, k, m_dst, n_dst, n_ext, n_u, n_v, n_lc = *n_lc_, rrs, rre; int32_t *f, *v, *t, li_qs, li_qe, li_rs, li_re, lj_qs, lj_qe, lj_rs, lj_re; int64_t *p; uint64_t *u; mg_path_dst_t *dst; gc_frag_t *a; mg_lchain_t *swap; // char *qs; asg_t *g = ug->g; *u_ = 0; if (n_lc == 0) return 0; // extend_lchain(lc, n_lc, qlen, ug); KMALLOC(km, a, n_lc); ///n_lc how many linear chains; just filter some linear chains for (i = n_ext = 0; i < n_lc; ++i) { // a[] is a view of frag[]; for sorting mg_lchain_t *r = &lc[i]; gc_frag_t *ai = &a[i]; int32_t is_isolated = 0, min_end_dist_g; transfor_coord(r, qlen, ug->u.a[r->v>>1].len, NULL, NULL, &rrs, &rre); r->dist_pre = -1;///indicate parent in graph chain min_end_dist_g = g->seq[r->v>>1].len - rre;///r->v: ref_id|rev if (rrs < min_end_dist_g) min_end_dist_g = rrs; if (min_end_dist_g > max_dist_g) is_isolated = 1; // if too far from segment ends else if (min_end_dist_g>>3 > r->score) is_isolated = 1; // if the lchain too small relative to distance to the segment ends ai->srt = (uint32_t)is_isolated<<31 | r->qe; ai->i = i; if (!is_isolated) ++n_ext; } ///if the alignment is too far from segment ends, which means it cannot contribute to graph alignment if (n_ext < 2) { // no graph chaining needed; early return kfree(km, a); KMALLOC(km, u, n_lc); for (i = 0; i < n_lc; ++i) u[i] = (uint64_t)lc[i].score<<32 | 1; *u_ = u; // compress_lchain(lc, n_lc, qlen, ug, an); return n_lc; } radix_sort_gc(a, a + n_lc);///sort by: is_isolated(1):qe KMALLOC(km, v, n_lc); KMALLOC(km, f, n_ext); KMALLOC(km, p, n_ext); KCALLOC(km, t, n_ext); // KMALLOC(km, qs, max_dist_q + 1);//for m_dst = n_dst = 0, dst = 0; ///n_ext is number of linear chains that might be included in graph chains ///sorted by the positions in query; sorted by qe of each chain for (i = 0; i < n_ext; ++i) { // core loop gc_frag_t *ai = &a[i]; mg_lchain_t *li = &lc[ai->i];///linear chain; sorted by qe, i.e. end position in query int32_t mm_ovlp = max_ovlp(ug->g, li->v^1); transfor_coord(li, qlen, ug->u.a[li->v>>1].len, &li_qs, &li_qe, &li_rs, &li_re); ///note segi is query id, instead of ref id; it is not such useful /** * a[].x: idx_in_minimizer_arr(32)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) **/ { // collect end points potentially reachable from _i_ int32_t x = li->qs + bw + mm_ovlp, n_skip = 0; if (x > qlen) x = qlen; ///collect alignments that can be reachable from the left side ///that is, a[x].qe <= x x = find_max(i, a, x); n_dst = 0; for (j = x; j >= 0; --j) { // collect potential destination vertices gc_frag_t *aj = &a[j]; //potential chains that might be overlapped with the left side of li mg_lchain_t *lj = &lc[aj->i]; mg_path_dst_t *q; int32_t target_dist, dq/**, so = specific_ovlp(ug, uopt, li->v^1, lj->v^1)**/; transfor_coord(lj, qlen, ug->u.a[lj->v>>1].len, &lj_qs, &lj_qe, &lj_rs, &lj_re); ///lj->qs >= li->qs && lj->qe <= li->qs, so lj is contained if (lj->qs >= li->qs) continue; // lj is contained in li on the query coordinate /** * doesn't work for overlap graph if (lj_qe > li_qs) { // test overlap on the query int o = lj_qe - li_qs - so;///get_lchain_ovlp(lj, li, ug->g, qlen, ug); ///mask_level = 0.5, if overlap is too long ///note here is the overlap in query, so too long overlaps might be wrong if (o > (lj->qe - lj->qs) * mask_level || o > (li->qe - li->qs) * mask_level) continue; } **/ dq = li_qs - lj_qe;///dq might be smaller than 0 if (dq > max_dist_q) break; // if query gap too large, stop ///The above filter chains like: ///1. lj is contained in li ///2. the overlap between li and lj is too large ///3. li and lj are too far ///above we have checked gap/overlap in query ///then we need to check gap/overlap in reference if (li->v != lj->v) { // the two linear chains are on two different refs // minimal graph gap; the real graph gap might be larger int32_t min_dist = li_rs + (g->seq[lj->v>>1].len - lj_re); if (min_dist > max_dist_g) continue; // graph gap too large //note here min_dist - (lj->qs - li->qe) > bw is important //min_dist is always larger than 0, (lj->qs - li->qe) might be negative /** * doesn't work for overlap graph min_dist -= so; if (min_dist - bw > li->qs - lj->qe) continue; ///note seg* is the query id, instead of ref id **/ target_dist = mg_target_dist(g, lj, li); if (target_dist < 0) continue; // this may happen if the query overlap is far too large } else if (lj->rs >= li->rs || lj->re >= li->re) { // not colinear continue; } else {///li->v == lj->v and colinear; at the same ref id ///w is indel, w is always positive int32_t dr = li->rs - lj->re, dq = li->qs - lj->qe, w = dr > dq? dr - dq : dq - dr; ///note that l*->v is the ref id, while seg* is the query id if (w > bw) continue; // test bandwidth if (dr > max_dist_g || dr < -max_dist_g) continue; if (lj->re > li->rs) { // test overlap on the graph segment int o = lj->re - li->rs; if (o > (lj->re - lj->rs) * mask_level || o > (li->re - li->rs) * mask_level) continue; } target_dist = mg_target_dist(g, lj, li); } if (n_dst == m_dst) KEXPAND(km, dst, m_dst); // TODO: watch out the quadratic behavior! q = &dst[n_dst++];///q saves information for i->j memset(q, 0, sizeof(mg_path_dst_t)); ///note v is (rid:rev), so two alignment chains might be at the same ref id with different directions q->inner = (li->v == lj->v); q->v = lj->v^1;///must be v^1 instead of v q->meta = j; q->qlen = li->qs - lj->qe;///might be negative /** * doesn't work for overlap graph q->so = 0; if(li->v != lj->v && lj->qe > li->qs) { lj_qe = lj->qe; li_qs = li->qs + g->seq[lj->v>>1].len - so; q->so = lj_qe - li_qs; if(q->so < 0) q->so = 0; } **/ q->target_dist = target_dist;///cannot understand the target_dist q->target_hash = 0; q->check_hash = 0; if (t[j] == i) {///this pre-cut is weird; attention if (++n_skip > max_skip) break; } if (p[j] >= 0) t[p[j]] = i; } } ///the above saves all linear chains that might be reached to the left side of chain i ///all those chains are saved to dst { // confirm reach-ability int32_t k; // test reach-ability without sequences // (g->seg[li->v>>1].len - li->rs) ----> is like the node length in string graph mg_shortest_k(km, g, li->v^1, n_dst, dst, max_dist_g + (g->seq[li->v>>1].len - li->rs), MG_MAX_SHORT_K, /**0, 0, 1,**/ 0); // remove unreachable destinations for (j = k = 0; j < n_dst; ++j) { mg_path_dst_t *dj = &dst[j]; int32_t sc; if (dj->n_path == 0) continue; // unreachable sc = cal_sc(dj, li, lc, an, a, f, bw, ref_bonus, chn_pen_gap); if (sc == INT32_MIN) continue; // out of band if (sc + li->score < 0) continue; // negative score and too low dst[k] = dst[j]; dst[k++].srt_key = INT64_MAX/2 - (int64_t)sc; // sort in the descending order } n_dst = k; if (n_dst > 0) { radix_sort_dst(dst, dst + n_dst); // discard weaker chains if the best score is much larger (assuming base-level heuristic won't lift it to the top chain) // dst[0].srt_key has the largest score for (j = 1; j < n_dst; ++j) if ((int64_t)(dst[j].srt_key - dst[0].srt_key) > li->score)//discard chains with too small weight break; n_dst = j; if (n_dst > max_gc_seq_ext) n_dst = max_gc_seq_ext; // discard weaker chains } } /** //path if (n_dst > 0) { // find paths with sequences int32_t min_qs = li->qs; for (j = 0; j < n_dst; ++j) { const mg_lchain_t *lj; assert(dst[j].n_path > 0); ///a[]->srt = (uint32_t)is_isolated<<31 | r->qe; ///a[]->i = i; lj = &lc[a[dst[j].meta].i]; if (lj->qe < min_qs) min_qs = lj->qe; } ///qs keeps the sequence at the gap between the li and lj in query memcpy(qs, &qseq[min_qs], li->qs - min_qs); mg_shortest_k(km, g, li->v^1, n_dst, dst, max_dist_g + (g->seg[li->v>>1].len - li->rs), MG_MAX_SHORT_K, li->qs - min_qs, qs, 1, 0); if (mg_dbg_flag & MG_DBG_GC1) fprintf(stderr, "[src:%d] q_intv=[%d,%d), src=%c%s[%d], n_dst=%d, max_dist=%d, min_qs=%d, lc_score=%d\n", ai->i, li->qs, li->qe, "><"[(li->v&1)^1], g->seg[li->v>>1].name, li->v^1, n_dst, max_dist_g + (g->seg[li->v>>1].len - li->rs), min_qs, li->score); }**/ { // DP int32_t max_f = li->score, max_j = -1, max_d = -1, max_inner = 0; uint32_t max_hash = 0; for (j = 0; j < n_dst; ++j) { mg_path_dst_t *dj = &dst[j]; int32_t sc; sc = cal_sc(dj, li, lc, an, a, f, bw, ref_bonus, chn_pen_gap); if (sc == INT32_MIN) continue; if (sc > max_f) max_f = sc, max_j = dj->meta, max_d = dj->dist, max_hash = dj->hash, max_inner = dj->inner; } f[i] = max_f, p[i] = max_j; li->dist_pre = max_d; li->hash_pre = max_hash; li->inner_pre = max_inner; v[i] = max_j >= 0 && v[max_j] > max_f? v[max_j] : max_f; } } kfree(km, dst); // print_gchain(a, p, lc, n_ext, ug, qlen); // kfree(km, qs); ///n_ext: number of useful chains ///n_lc - n_ext: number of isoated chains u = mg_chain_backtrack(km, n_ext, f, p, v, t, 0, 0, n_lc - n_ext, &n_u, &n_v); kfree(km, f); kfree(km, p); kfree(km, t); ///store the extra isoated chains for (i = 0; i < n_lc - n_ext; ++i) { u[n_u++] = (uint64_t)lc[a[n_ext + i].i].score << 32 | 1; v[n_v++] = n_ext + i; } ///reorganize lc; KMALLOC(km, swap, n_v); for (i = 0, k = 0; i < n_u; ++i) { int32_t k0 = k, ni = (int32_t)u[i]; for (j = 0; j < ni; ++j) swap[k++] = lc[a[v[k0 + (ni - j - 1)]].i]; } assert(k == n_v); memcpy(lc, swap, n_v * sizeof(mg_lchain_t)); *n_lc_ = n_v; *u_ = u; // compress_lchain(lc, *n_lc_, qlen, ug, an); kfree(km, a); kfree(km, swap); kfree(km, v); return n_u; } static inline void copy_lchain(mg_llchain_t *q, const mg_lchain_t *p, int32_t *n_a, mg128_t *a_new, const mg128_t *a_old) { q->cnt = p->cnt, q->v = p->v, q->score = p->score; memcpy(&a_new[*n_a], &a_old[p->off], q->cnt * sizeof(mg128_t)); q->off = *n_a; (*n_a) += q->cnt; } void mg_gchain_extra(const asg_t *g, mg_gchains_t *gs) { int32_t i, j, k; for (i = 0; i < gs->n_gc; ++i) { // iterate over gchains mg_gchain_t *p = &gs->gc[i]; const mg_llchain_t *q; const mg128_t *last_a; int32_t q_span, rest_pl, tmp, n_mini; p->qs = p->qe = p->ps = p->pe = -1, p->plen = p->blen = p->mlen = 0, p->div = -1.0f; if (p->cnt == 0) continue; ///some linear chains in middle might be [].cnt == 0 ///but for the first and the last linear chains, [].cnt > 0 assert(gs->lc[p->off].cnt > 0 && gs->lc[p->off + p->cnt - 1].cnt > 0); // first and last lchains can't be empty q = &gs->lc[p->off]; q_span = (int32_t)(gs->a[q->off].y>>32&0xff); /** * a[].x: idx_in_minimizer_arr(32)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) * **/ p->qs = (int32_t)gs->a[q->off].y + 1 - q_span;///calculated by the first lchain p->ps = (int32_t)gs->a[q->off].x + 1 - q_span;///calculated by the first lchain tmp = (int32_t)(gs->a[q->off].x>>32); assert(p->qs >= 0 && p->ps >= 0); q = &gs->lc[p->off + p->cnt - 1];///last lchain p->qe = (int32_t)gs->a[q->off + q->cnt - 1].y + 1; p->pe = g->seq[q->v>>1].len - (int32_t)gs->a[q->off + q->cnt - 1].x - 1; // this is temporary n_mini = (int32_t)(gs->a[q->off + q->cnt - 1].x>>32) - tmp + 1; assert(p->n_anchor > 0); rest_pl = 0; // this value is never used if the first lchain is not empty (which should always be true) last_a = &gs->a[gs->lc[p->off].off];///first minizers in the first linear chain for (j = 0; j < p->cnt; ++j) { // iterate over lchains const mg_llchain_t *q = &gs->lc[p->off + j]; int32_t vlen = g->seq[q->v>>1].len;///node length in graph p->plen += vlen; for (k = 0; k < q->cnt; ++k) { // iterate over anchors const mg128_t *r = &gs->a[q->off + k]; int32_t pl, ql = (int32_t)r->y - (int32_t)last_a->y; int32_t span = (int32_t)(r->y>>32&0xff); if (j == 0 && k == 0) { // the first anchor on the first lchain pl = ql = span; } else if (j > 0 && k == 0) { // the first anchor but not on the first lchain pl = (int32_t)r->x + 1 + rest_pl; } else { pl = (int32_t)r->x - (int32_t)last_a->x; } if (ql < 0) ql = -ql, n_mini += (int32_t)(last_a->x>>32) - (int32_t)(r->x>>32); // dealing with overlapping query at junctions p->blen += pl > ql? pl : ql; p->mlen += pl > span && ql > span? span : pl < ql? pl : ql; last_a = r; } if (q->cnt == 0) rest_pl += vlen; else rest_pl = vlen - (int32_t)gs->a[q->off + q->cnt - 1].x - 1; } p->pe = p->plen - p->pe; assert(p->pe >= p->ps); // here n_mini >= p->n_anchor should stand almost all the time p->div = n_mini >= p->n_anchor? log((double)n_mini / p->n_anchor) / q_span : log((double)p->n_anchor / n_mini) / q_span; } } // reorder gcs->a[] and gcs->lc[] such that they are in the same order as gcs->gc[] void mg_gchain_restore_order(void *km, mg_gchains_t *gcs) { int32_t i, n_a, n_lc; mg_llchain_t *lc; mg128_t *a; KMALLOC(km, lc, gcs->n_lc); KMALLOC(km, a, gcs->n_a); n_a = n_lc = 0; for (i = 0; i < gcs->n_gc; ++i) { mg_gchain_t *gc = &gcs->gc[i]; assert(gc->cnt > 0); memcpy(&lc[n_lc], &gcs->lc[gc->off], gc->cnt * sizeof(mg_llchain_t)); memcpy(&a[n_a], &gcs->a[gcs->lc[gc->off].off], gc->n_anchor * sizeof(mg128_t)); n_lc += gc->cnt, n_a += gc->n_anchor; } memcpy(gcs->lc, lc, gcs->n_lc * sizeof(mg_llchain_t)); memcpy(gcs->a, a, gcs->n_a * sizeof(mg128_t)); kfree(km, lc); kfree(km, a); } // sort chains by score void mg_gchain_sort_by_score(void *km, mg_gchains_t *gcs) { mg128_t *z; mg_gchain_t *gc; int32_t i; KMALLOC(km, z, gcs->n_gc); KMALLOC(km, gc, gcs->n_gc); for (i = 0; i < gcs->n_gc; ++i) z[i].x = (uint64_t)gcs->gc[i].score << 32 | gcs->gc[i].hash, z[i].y = i; radix_sort_128x(z, z + gcs->n_gc); for (i = gcs->n_gc - 1; i >= 0; --i) gc[gcs->n_gc - 1 - i] = gcs->gc[z[i].y]; memcpy(gcs->gc, gc, gcs->n_gc * sizeof(mg_gchain_t)); kfree(km, z); kfree(km, gc); mg_gchain_restore_order(km, gcs); // this put gcs in the proper order } ///u[]: sc|occ of chains ///a[]: candidate list ///gcs[0] = mg_gchain_gen(0, b->km, gi->g, n_gc, u, lc, a, hash, opt->min_gc_cnt, opt->min_gc_score); // TODO: if frequent malloc() is a concern, filter first and then generate gchains; or generate gchains in thread-local pool and then move to global malloc() mg_gchains_t *mg_gchain_gen(void *km_dst, void *km, const asg_t *g, int32_t n_u, const uint64_t *u, const mg_lchain_t *lc, const mg128_t *a, uint32_t hash, int32_t min_gc_cnt, int32_t min_gc_score) { mg_gchains_t *gc; mg_llchain_t *tmp; int32_t i, j, k, st, n_g, n_a, s_tmp, n_tmp, m_tmp; KCALLOC(km_dst, gc, 1); // count the number of gchains and remaining anchors // filter out low-quality g_chains for (i = 0, st = 0, n_g = n_a = 0; i < n_u; ++i) { ///nui: how many linear chaisn in i-th g_chain int32_t m = 0, nui = (int32_t)u[i]; for (j = 0; j < nui; ++j) m += lc[st + j].cnt; // m is the number of anchors in this gchain if (m >= min_gc_cnt && (int64_t)(u[i]>>32) >= min_gc_score) ++n_g, n_a += m; st += nui; } if (n_g == 0) return gc; // preallocate gc->km = km_dst; gc->n_gc = n_g, gc->n_a = n_a; KCALLOC(km_dst, gc->gc, n_g);///all graph chains KMALLOC(km_dst, gc->a, n_a);///all anchors, aka minimizers // core loop tmp = 0; s_tmp = n_tmp = m_tmp = 0; for (i = k = 0, st = 0, n_a = 0; i < n_u; ++i) { int32_t n_a0 = n_a, m = 0, nui = (int32_t)u[i]; ///nui: how many linear chaisn in i-th g_chain for (j = 0; j < nui; ++j) m += lc[st + j].cnt; ///how many minizers in i-th g_chain if (m >= min_gc_cnt && (int64_t)(u[i]>>32) >= min_gc_score) { mg_llchain_t *q; uint32_t h = hash; gc->gc[k].score = u[i]>>32; ///chain score gc->gc[k].off = s_tmp; ///all minimizers of k-th chain: gc->a[gc->gc[k].off, ) for (j = 0; j < nui; ++j) {///how many linear chains const mg_lchain_t *p = &lc[st + j]; h += __ac_Wang_hash(p->qs) + __ac_Wang_hash(p->re) + __ac_Wang_hash(p->v); } gc->gc[k].hash = __ac_Wang_hash(h);///hash key for the k-th graph chain if (n_tmp == m_tmp) KEXPAND(km, tmp, m_tmp); // copy the first lchain to gc->a[] and tmp[] (aka, gc->lc[]) // for the first lchain, it is easy and we just copy all its anchors copy_lchain(&tmp[n_tmp++], &lc[st], &n_a, gc->a, a); ///0-th lchain has been stored ///process the remaining chains for (j = 1; j < nui; ++j) { const mg_lchain_t *l0 = &lc[st + j - 1], *l1 = &lc[st + j]; if (!l1->inner_pre) { // bridging two segments; if l0 and l1 are at different reference int32_t s, n_pathv; mg_path_dst_t dst; mg_pathv_t *p; memset(&dst, 0, sizeof(mg_path_dst_t)); dst.v = l0->v ^ 1; assert(l1->dist_pre >= 0); dst.target_dist = l1->dist_pre; dst.target_hash = l1->hash_pre;///hash value of the whole path dst.check_hash = 1; p = mg_shortest_k(km, g, l1->v^1, 1, &dst, dst.target_dist, MG_MAX_SHORT_K, &n_pathv); if (n_pathv == 0 || dst.target_hash != dst.hash) fprintf(stderr, "%c[%d] -> %c[%d], dist=%d, target_dist=%d\n", "><"[(l1->v^1)&1], l1->v^1, "><"[(l0->v^1)&1], l0->v^1, dst.dist, dst.target_dist); assert(n_pathv > 0); assert(dst.target_hash == dst.hash); for (s = n_pathv - 2; s >= 1; --s) { // path found in a backward way, so we need to reverse it if (n_tmp == m_tmp) KEXPAND(km, tmp, m_tmp); q = &tmp[n_tmp++]; q->off = q->cnt = q->score = 0; q->v = p[s].v^1; // when reversing a path, we also need to flip the orientation } kfree(km, p); if (n_tmp == m_tmp) KEXPAND(km, tmp, m_tmp); copy_lchain(&tmp[n_tmp++], l1, &n_a, gc->a, a); } else { // if both of them are at the same linear chain, just merge them #if 1 int32_t k; mg_llchain_t *t = &tmp[n_tmp - 1];//the last lchain, have alread done assert(l0->v == l1->v); // a[].x: ref_id(31)rev(1)r_pos(32) // a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) for (k = 0; k < l1->cnt; ++k) { const mg128_t *ak = &a[l1->off + k]; if ((int32_t)ak->x > l0->re && (int32_t)ak->y > l0->qe)//find colinear anchors break; } assert(k < l1->cnt); t->cnt += l1->cnt - k, t->score += l1->score; memcpy(&gc->a[n_a], &a[l1->off + k], (l1->cnt - k) * sizeof(mg128_t)); n_a += l1->cnt - k; #else // don't use this block; for debugging only if (n_tmp == m_tmp) KEXPAND(km, tmp, m_tmp); copy_lchain(&tmp[n_tmp++], l1, &n_a, gc->a, a); #endif } } gc->gc[k].cnt = n_tmp - s_tmp; gc->gc[k].n_anchor = n_a - n_a0; ++k, s_tmp = n_tmp; } st += nui;//nui: how many linear chains in this gchain } assert(n_a <= gc->n_a); gc->n_a = n_a; gc->n_lc = n_tmp; KMALLOC(km_dst, gc->lc, n_tmp); memcpy(gc->lc, tmp, n_tmp * sizeof(mg_llchain_t)); kfree(km, tmp); mg_gchain_extra(g, gc); mg_gchain_sort_by_score(km, gc); return gc; } // set r[].{id,parent,subsc}, ASSUMING r[] is sorted by score // mg_gchain_set_parent(b->km, opt->mask_level, gcs[0]->n_gc, gcs[0]->gc, opt->sub_diff, 0); void mg_gchain_set_parent(void *km, float mask_level, int n, mg_gchain_t *r, int sub_diff, int hard_mask_level) { int i, j, k, *w; uint64_t *cov; if (n <= 0) return; for (i = 0; i < n; ++i) r[i].id = i; cov = (uint64_t*)kmalloc(km, n * sizeof(uint64_t)); w = (int*)kmalloc(km, n * sizeof(int)); w[0] = 0, r[0].parent = 0;///the first gchain is a primary hits; since all gchains have already been sorted by scores for (i = 1, k = 1; i < n; ++i) {///start from the 1-th chain, instead of the 0-th chain mg_gchain_t *ri = &r[i]; int si = ri->qs, ei = ri->qe, n_cov = 0, uncov_len = 0; if (hard_mask_level) goto skip_uncov; for (j = 0; j < k; ++j) { // traverse existing primary hits to find overlapping hits mg_gchain_t *rp = &r[w[j]]; int sj = rp->qs, ej = rp->qe; if (ej <= si || sj >= ei) continue;///no overlaps if (sj < si) sj = si;///MAX(si, sj) if (ej > ei) ej = ei;///MIN(ei, ej) cov[n_cov++] = (uint64_t)sj<<32 | ej;///overlap coordinates } if (n_cov == 0) { goto set_parent_test; // no overlapping primary hits; then i is a new primary hit } else if (n_cov > 0) { // there are overlapping primary hits; find the length not covered by existing primary hits int j, x = si; radix_sort_gfa64(cov, cov + n_cov); for (j = 0; j < n_cov; ++j) { if ((int)(cov[j]>>32) > x) uncov_len += (cov[j]>>32) - x; x = (int32_t)cov[j] > x? (int32_t)cov[j] : x; } if (ei > x) uncov_len += ei - x; } skip_uncov: for (j = 0; j < k; ++j) { // traverse existing primary hits again mg_gchain_t *rp = &r[w[j]]; int sj = rp->qs, ej = rp->qe, min, max, ol; if (ej <= si || sj >= ei) continue; // no overlap min = ej - sj < ei - si? ej - sj : ei - si;///chain length max = ej - sj > ei - si? ej - sj : ei - si;///chain length ol = si < sj? (ei < sj? 0 : ei < ej? ei - sj : ej - sj) : (ej < si? 0 : ej < ei? ej - si : ei - si); // overlap length; TODO: this can be simplified if ((float)ol / min - (float)uncov_len / max > mask_level) { int cnt_sub = 0; ri->parent = rp->parent; rp->subsc = rp->subsc > ri->score? rp->subsc : ri->score; if (ri->cnt >= rp->cnt) cnt_sub = 1; if (cnt_sub) ++rp->n_sub; break; } } set_parent_test: if (j == k) w[k++] = i, ri->parent = i, ri->n_sub = 0; } kfree(km, cov); kfree(km, w); } // set r[].flt, i.e. mark weak suboptimal chains as filtered int mg_gchain_flt_sub(float pri_ratio, int min_diff, int best_n, int n, mg_gchain_t *r) { if (pri_ratio > 0.0f && n > 0) { int i, k, n_2nd = 0; for (i = k = 0; i < n; ++i) { int p = r[i].parent; if (p == i) { // primary r[i].flt = 0, ++k; } else if ((r[i].score >= r[p].score * pri_ratio || r[i].score + min_diff >= r[p].score) && n_2nd < best_n) { if (!(r[i].qs == r[p].qs && r[i].qe == r[p].qe && r[i].ps == r[p].ps && r[i].pe == r[p].pe)) // not identical hits; TODO: check path as well r[i].flt = 0, ++n_2nd, ++k; else r[i].flt = 1; } else r[i].flt = 1; } return k; } return n; } // recompute gcs->gc[].{off,n_anchor} and gcs->lc[].off, ASSUMING they are properly ordered (see mg_gchain_restore_order) void mg_gchain_restore_offset(mg_gchains_t *gcs) { int32_t i, j, n_a, n_lc; for (i = 0, n_a = n_lc = 0; i < gcs->n_gc; ++i) { mg_gchain_t *gc = &gcs->gc[i]; gc->off = n_lc; for (j = 0, gc->n_anchor = 0; j < gc->cnt; ++j) { mg_llchain_t *lc = &gcs->lc[n_lc + j]; lc->off = n_a; n_a += lc->cnt; gc->n_anchor += lc->cnt; } n_lc += gc->cnt; } assert(n_lc == gcs->n_lc && n_a == gcs->n_a); } // hard drop filtered chains, ASSUMING gcs is properly ordered void mg_gchain_drop_flt(void *km, mg_gchains_t *gcs) { int32_t i, n_gc, n_lc, n_a, n_lc0, n_a0, *o2n; if (gcs->n_gc == 0) return; KMALLOC(km, o2n, gcs->n_gc); for (i = 0, n_gc = 0; i < gcs->n_gc; ++i) { mg_gchain_t *r = &gcs->gc[i]; o2n[i] = -1; if (r->flt || r->cnt == 0) continue; o2n[i] = n_gc++; } n_gc = n_lc = n_a = 0; n_lc0 = n_a0 = 0; for (i = 0; i < gcs->n_gc; ++i) { mg_gchain_t *r = &gcs->gc[i]; if (o2n[i] >= 0) { memmove(&gcs->a[n_a], &gcs->a[n_a0], r->n_anchor * sizeof(mg128_t)); memmove(&gcs->lc[n_lc], &gcs->lc[n_lc0], r->cnt * sizeof(mg_llchain_t)); gcs->gc[n_gc] = *r; gcs->gc[n_gc].id = n_gc; gcs->gc[n_gc].parent = o2n[gcs->gc[n_gc].parent]; ++n_gc, n_lc += r->cnt, n_a += r->n_anchor; } n_lc0 += r->cnt, n_a0 += r->n_anchor; } assert(n_lc0 == gcs->n_lc && n_a0 == gcs->n_a); kfree(km, o2n); gcs->n_gc = n_gc, gcs->n_lc = n_lc, gcs->n_a = n_a; if (n_a != n_a0) { KREALLOC(gcs->km, gcs->a, gcs->n_a); KREALLOC(gcs->km, gcs->lc, gcs->n_lc); KREALLOC(gcs->km, gcs->gc, gcs->n_gc); } mg_gchain_restore_offset(gcs); } // estimate mapping quality ///mg_gchain_set_mapq(b->km, gcs, qlen, mz->n, opt->min_gc_score); void mg_gchain_set_mapq(void *km, mg_gchains_t *gcs, int qlen, int max_mini, int min_gc_score) { static const float q_coef = 40.0f; int64_t sum_sc = 0; float uniq_ratio, r_sc, r_cnt; int i, t_sc, t_cnt; if (gcs == 0 || gcs->n_gc == 0) return; t_sc = qlen < 100? qlen : 100; t_cnt = max_mini < 10? max_mini : 10; if (t_cnt < 5) t_cnt = 5; r_sc = 1.0 / t_sc; r_cnt = 1.0 / t_cnt; for (i = 0; i < gcs->n_gc; ++i) if (gcs->gc[i].parent == gcs->gc[i].id) sum_sc += gcs->gc[i].score;///primary chain uniq_ratio = (float)sum_sc / (sum_sc + gcs->rep_len); for (i = 0; i < gcs->n_gc; ++i) { mg_gchain_t *r = &gcs->gc[i]; if (r->parent == r->id) {///primary chain int mapq, subsc; float pen_s1 = (r->score > t_sc? 1.0f : r->score * r_sc) * uniq_ratio; float x, pen_cm = r->n_anchor > t_cnt? 1.0f : r->n_anchor * r_cnt; pen_cm = pen_s1 < pen_cm? pen_s1 : pen_cm; subsc = r->subsc > min_gc_score? r->subsc : min_gc_score; x = (float)subsc / r->score; mapq = (int)(pen_cm * q_coef * (1.0f - x) * logf(r->score)); mapq -= (int)(4.343f * logf(r->n_sub + 1) + .499f); mapq = mapq > 0? mapq : 0; if (r->score > subsc && mapq == 0) mapq = 1; r->mapq = mapq < 60? mapq : 60; } else r->mapq = 0; } } void mg_map_frag(const void *ha_flt_tab, const ha_pt_t *ha_idx, const ma_ug_t *ug, const asg_t *rg, const uint32_t qid, const int qlen, const char *qseq, ha_mzl_v *mz, st_mt_t *sp, mg_tbuf_t *b, int32_t w, int32_t k, int32_t hpc, int32_t mz_sd, int32_t mz_rewin, const mg_idxopt_t *opt, const ug_opt_t *uopt, mg_gchains_t **gcs) { mg128_t *a = NULL; int64_t n_a; int32_t *mini_pos; int i, rep_len, n_mini_pos, n_lc, max_chain_gap_qry, max_chain_gap_ref, n_gc; uint32_t hash; uint64_t *u; mg_lchain_t *lc; km_stat_t kmst; (*gcs) = NULL; hash = qid; hash ^= __ac_Wang_hash(qlen) + __ac_Wang_hash(opt->seed); hash = __ac_Wang_hash(hash); mz->n = 0; mz2_ha_sketch(qseq, qlen, w, k, 0, hpc, mz, ha_flt_tab, mz_sd, NULL, NULL, NULL, -1, -1, -1, sp, mz_rewin, 1, NULL); ///a[]->y: weight(8)seg_id(8)flag(8)span(8)pos(32);--->query ///a[]->x: rid(31)rev(1)rpos(33);--->reference a = collect_seed_hits(b->km, opt, 1/**opt->hap_n**/, ha_flt_tab, ha_idx, ug, mz, &n_a, &rep_len, &n_mini_pos, &mini_pos); /** // might be recover if (opt->max_gap_pre > 0 && opt->max_gap_pre * 2 < opt->max_gap) n_a = flt_anchors(n_a, a, opt->max_gap_pre); max_chain_gap_qry = max_chain_gap_ref = opt->max_gap; **/ max_chain_gap_qry = max_chain_gap_ref = qlen*2; if (n_a == 0) {//no matched minimizer if(a) kfree(b->km, a); a = 0, n_lc = 0, u = 0; } else { a = mg_lchain_dp(max_chain_gap_ref, max_chain_gap_qry, opt->bw, opt->max_lc_skip, opt->max_lc_iter, opt->min_lc_cnt, opt->min_lc_score, opt->chn_pen_gap, n_a, a, &n_lc, &u, b->km); } if (n_lc) {///n_lc is how many linear chain we found lc = mg_lchain_gen(b->km, qlen, n_lc, u, a, ug);//lc->the status of each chain; u->idx of each chain; for (i = 0; i < n_lc; ++i)///update a[] since ref_id|rev has already been saved to lc[].v mg_update_anchors(lc[i].cnt, &a[lc[i].off], n_mini_pos, mini_pos);///update a[].x } else lc = 0; kfree(b->km, mini_pos); kfree(b->km, u); // fprintf(stderr, "++0++qid: %u, qlen: %d, n_a: %ld, n_lc: %d\n", qid, qlen, n_a, n_lc); /** * up to here, a[] has been changed * a[].x: idx_in_minimizer_arr(32)r_pos(32) * a[].y: weight(8)query_id(8)flag(8)span(8)q_pos(32) **/ // for (i = 0; i < n_lc; i++) { // mg_lchain_t *ri = &lc[i]; // fprintf(stderr, "+0)))))))))))))))))))))))))))+\tA\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%d\tts:%u\tte:%u\ttl:%u\n", // (ri->v>>1)+1, "lc"[ug->u.a[ri->v>>1].circ], "+-"[ri->v&1], ri->qs, ri->qe, qlen, ri->rs, ri->re, ug->u.a[ri->v>>1].len); // } max_chain_gap_qry = max_chain_gap_ref = opt->max_gap; n_gc = mg_gchain1_dp(b->km, ug, rg, &n_lc, lc, qlen, max_chain_gap_ref, max_chain_gap_qry, opt->bw, opt->max_gc_skip, opt->ref_bonus, opt->chn_pen_gap, opt->mask_level, opt->max_gc_seq_ext, uopt, a, &u); // for (i = 0; i < n_lc; i++) { // mg_lchain_t *ri = &lc[i]; // fprintf(stderr, "-0-\tA\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%d\tts:%u\tte:%u\ttl:%u\n", // (ri->v>>1)+1, "lc"[ug->u.a[ri->v>>1].circ], "+-"[ri->v&1], ri->qs, ri->qe, qlen, ri->rs, ri->re, ug->u.a[ri->v>>1].len); // } (*gcs) = mg_gchain_gen(0, b->km, ug->g, n_gc, u, lc, a, hash, opt->min_gc_cnt, opt->min_gc_score); (*gcs)->rep_len = rep_len; (*gcs)->qid = qid; (*gcs)->qlen = qlen; kfree(b->km, a); kfree(b->km, lc); kfree(b->km, u); mg_gchain_set_parent(b->km, opt->mask_level, (*gcs)->n_gc, (*gcs)->gc, opt->sub_diff, 0); mg_gchain_flt_sub(opt->pri_ratio, k * 2, opt->best_n, (*gcs)->n_gc, (*gcs)->gc); mg_gchain_drop_flt(b->km, (*gcs)); mg_gchain_set_mapq(b->km, (*gcs), qlen, mz->n, opt->min_gc_score); if (b->km) { km_stat(b->km, &kmst); if (kmst.n_blocks != kmst.n_cores) { fprintf(stderr, "[E::%s] memory leak at %u\n", __func__, qid); abort(); } if (kmst.largest > 1U<<28) { km_destroy(b->km); b->km = km_init(); } } // fprintf(stderr, "++6++qid: %u, (*gcs)->n_gc: %d\n", qid, (*gcs)->n_gc); } static void worker_for_ul_alignment(void *data, long i, int tid) // callback for kt_for() { utepdat_t *s = (utepdat_t*)data; mg_map_frag(s->ha_flt_tab, s->ha_idx, s->ug, s->rg, s->id+i, s->len[i], s->seq[i], &(s->mzs[tid]), &(s->sps[tid]), s->buf[tid], s->opt->w, s->opt->k, s->opt->is_HPC, asm_opt.mz_sample_dist, asm_opt.mz_rewin, s->opt, s->uopt, &(s->gcs[i])); } uint32_t overlap_statistics(overlap_region_alloc* olist, ma_ug_t *ug, int64_t *tt, uint8_t mm) { uint32_t k, sp = (uint32_t)-1, ep = (uint32_t)-1, l = 0; for (k = 0; k < olist->length; k++) { /** if(b->olist.list[k].y_id != 38) continue; **/ /** for (z = 0, te = ta = tua = 0; z < b->olist.list[k].w_list_length; z++) { if(b->olist.list[k].w_list[z].y_end != -1) { te += b->olist.list[k].w_list[z].error; ta += b->olist.list[k].w_list[z].x_end + 1 - b->olist.list[k].w_list[z].x_start; fprintf(stderr, "x->[%lu, %lu), y->[%d, %d), e->%d\n", b->olist.list[k].w_list[z].x_start, b->olist.list[k].w_list[z].x_end+1, b->olist.list[k].w_list[z].y_start, b->olist.list[k].w_list[z].y_end+1, b->olist.list[k].w_list[z].error); } else { tua += b->olist.list[k].w_list[z].x_end + 1 - b->olist.list[k].w_list[z].x_start; } } fprintf(stderr, "[M::utg%.6d%c::is_match:%u] x->[%u, %u); y->[%u, %u), ualigned->%u, e_rate->%f\n", b->olist.list[k].y_id+1, "lc"[s->ug->u.a[b->olist.list[k].y_id].circ], b->olist.list[k].is_match == 1, b->olist.list[k].x_pos_s, b->olist.list[k].x_pos_e+1, b->olist.list[k].y_pos_s, b->olist.list[k].y_pos_e+1, tua, (float)te/(float)ta); **/ if(tt){ uint32_t z; for (z = 0; z < olist->list[k].w_list_length; z++) { if(olist->list[k].w_list[z].y_end != -1) { if(tt) *tt += olist->list[k].w_list[z].x_end+1-olist->list[k].w_list[z].x_start; } } } if(olist->list[k].is_match == mm) { if(sp == (uint32_t)-1 || ep < olist->list[k].x_pos_s) { if(sp != (uint32_t)-1) l += ep + 1 - sp; sp = olist->list[k].x_pos_s; ep = olist->list[k].x_pos_e; } else { ep = MAX(ep, olist->list[k].x_pos_e); } if(ug) { fprintf(stderr, "[M::utg%.6d%c::is_match->%u] rev->%u, x->[%u, %u), y->[%u, %u)\n", (int)olist->list[k].y_id+1, "lc"[ug->u.a[olist->list[k].y_id].circ], olist->list[k].is_match, olist->list[k].y_pos_strand, olist->list[k].x_pos_s, olist->list[k].x_pos_e+1, olist->list[k].y_pos_s, olist->list[k].y_pos_e+1); } } } if(sp != (uint32_t)-1) l += ep + 1 - sp; return l; } /** void replace_ul(overlap_region_alloc* olist, Correct_dumy* dumy, haplotype_evdience_alloc* hap, const ul_idx_t *uu) { int64_t k, z, n = 0, c_qs, c_qe, c_ts, c_te, c_rev, p_qs, p_qe, p_te, p_ts, p_rev; uint64_t *sc = NULL, *track = NULL; overlap_region *c = NULL, *p = NULL; dumy->length = 0; for (k = 0; k < olist->length; k++) {///has already sorted by x_pos_e if(olist->list[k].is_match!=1) continue; dumy->overlapID[dumy->length] = (uint64_t)-1; dumy->overlapID[dumy->length] <<= 32; dumy->overlapID[dumy->length] |= k; dumy->length++; } kv_resize(uint64_t, hap->snp_srt, dumy->length); hap->snp_srt.n = dumy->length; memset(hap->snp_srt.a, 0, hap->snp_srt.n*sizeof(uint64_t)); sc = dumy->overlapID; track = hap->snp_srt.a; n = dumy->length; for (k = 0; k < n; k++) { c = &(olist->list[(uint32_t)track[k]]); for (z = k-1; z >= 0; z--) { p = &(olist->list[(uint32_t)track[z]]); } } } **/ uint64_t gl_chain_gen(overlap_region_alloc* olist, const ul_idx_t *uref, kv_ul_ov_t *res, uint32_t rec_trans, void *km) { uint64_t k, o2 = 0; ul_ov_t *p = NULL; res->n = 0; for (k = 0; k < olist->length; k++) { if(olist->list[k].is_match==2) o2++; if((!rec_trans) && olist->list[k].is_match!=1) continue; if(rec_trans && olist->list[k].is_match!=1 && olist->list[k].is_match!=2) continue; kv_pushp_km(km, ul_ov_t, *res, &p); p->qn = k/**olist->list[k].x_id**/; p->qs = olist->list[k].x_pos_s; p->qe = olist->list[k].x_pos_e+1; p->tn = olist->list[k].y_id; p->sec = 0; p->rev = olist->list[k].y_pos_strand; p->el = (olist->list[k].is_match==1?1:0); if(p->rev) { p->ts = uref->ug->u.a[p->tn].len - (olist->list[k].y_pos_e+1); p->te = uref->ug->u.a[p->tn].len - olist->list[k].y_pos_s; } else { p->ts = olist->list[k].y_pos_s; p->te = olist->list[k].y_pos_e+1; } } return o2; } int32_t find_ul_ov_max(int32_t n, const ul_ov_t *a, uint32_t x) { int32_t s = 0, e = n; if (n == 0) return -1; if (a[n-1].qe < x) return n - 1; if (a[0].qe >= x) return -1; while (e > s) { // TODO: finish this block int32_t m = s + (e - s) / 2; if (a[m].qe >= x) e = m; else s = m + 1; } assert(s == e); return s; } int64_t get_ecov(const ul_idx_t *uref, ul_ov_t *lv, ul_ov_t *lw, int64_t qlen, int64_t bw, double diff_ec_ul) { int64_t dis_q = lv->qe - lw->qe, dis_t = 0, dif, mm; uint32_t i, v = ((lv->tn<<1)|lv->rev)^1, w = ((lw->tn<<1)|lw->rev)^1; const asg_t *g = uref->ug->g; uint32_t nv = asg_arc_n(g, v); asg_arc_t *av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].del || av[i].v != w) continue; dis_t = ((uint32_t)av[i].ul); dis_t -= (lv->rev?lv->ts:g->seq[v>>1].len-lv->te); break; } dif = (dis_q>dis_t? dis_q-dis_t:dis_t-dis_q); mm = MAX(dis_q, dis_t); mm *= diff_ec_ul; if(mm < bw) mm = bw; // if((v>>1) == 1163 && (w>>1) == 1168) fprintf(stderr, ">>>>>>dis_q:%ld, dis_t:%ld, dif:%ld, mm:%ld\n", dis_q, dis_t, dif, mm); if(dif <= mm) return 1; return 0; } int64_t gl_exact_chain(kv_ul_ov_t *res, kv_ul_ov_t *ex, const ul_idx_t *uref, int64_t bw, double diff_ec_ul, int64_t qlen, uint64_t *srt, uint64_t *idx, uint64_t *track, void *km) { // fprintf(stderr, "*****************\n"); uint32_t li_v, lj_v; int64_t mm_ovlp, x, i, j, k, sc, csc, mm_sc, mm_idx; ul_ov_t *li = NULL, *lj = NULL; const asg_t *g = uref->ug->g; radix_sort_ul_ov_srt_qe(res->a, res->a + res->n); for (i = 0; i < (int64_t)res->n; ++i) { li = &(res->a[i]); li_v = (li->tn<<1)|li->rev; mm_ovlp = max_ovlp(g, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, res->a, x); csc = retrieve_u_cov_region(uref, li->tn, 0, li->ts, li->te, NULL); mm_sc = csc; mm_idx = -1; // fprintf(stderr, "---i:%ld, csc:%ld, li->tn:%u, li->ts:%u, li->te:%u\n", i, csc, li->tn, li->ts, li->te); for (j = x; j >= 0; --j) { // collect potential destination vertices lj = &(res->a[j]); lj_v = (lj->tn<<1)|lj->rev; // if(lj->qs >= li->qs) continue; // lj is contained in li on the query coordinate if(li_v != lj_v && get_ecov(uref, li, lj, qlen, bw, diff_ec_ul)) { sc = csc + (track[j]>>32); if(sc > mm_sc) mm_sc = sc, mm_idx = j; } } // 4294967295L track[i] = mm_sc; track[i] <<= 32; track[i] |= (mm_idx>=0?mm_idx:((uint64_t)0x7FFFFFFF)); srt[i] = mm_sc; srt[i] <<= 32; srt[i] |= i; // fprintf(stderr, "+++i:%ld, mm_idx:%ld, mm_sc:%ld\n", i, mm_idx, mm_sc); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n\n", li->tn+1, "lc"[uref->ug->u.a[li->tn].circ], li->qs, li->qe); } int64_t n_v, n_u, n_v0; radix_sort_gfa64(srt, srt+res->n); ex->n = res->n; for (k = (int64_t)res->n-1, n_v = n_u = 0; k >= 0; --k) { // fprintf(stderr, "\nk:%ld\n", k); n_v0 = n_v; for (i = (uint32_t)srt[k]; i >= 0 && (track[i]&((uint64_t)0x80000000)) == 0;) { ex->a[n_v++] = res->a[i]; track[i] |= ((uint64_t)0x80000000); // fprintf(stderr, "+i:%ld, ", i); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n", res->a[i].tn+1, "lc"[uref->ug->u.a[res->a[i].tn].circ], res->a[i].qs, res->a[i].qe); if((track[i]&((uint64_t)0x7FFFFFFF)) == ((uint64_t)0x7FFFFFFF)) i = -1; else i = track[i]&((uint64_t)0x7FFFFFFF); // if(i>=(int64_t)res->n) fprintf(stderr, "ERROR->i:%ld, res->n:%d, n_v:%ld, qlen:%ld\n", i, (int32_t)res->n, n_v, qlen); // fprintf(stderr, "next_i:%ld\n", i); // i = (olist->list[i].y_id == (uint32_t)-1?-1:olist->list[i].y_id); // fprintf(stderr, "-i:%ld\n", i); } if(n_v0 == n_v) continue; ///keep the whole score; do not cut score like minigraph // sc = (i<0?(srt[k]>>32):((srt[k]>>32)-olist->list[i].x_id)); sc = srt[k]>>32; idx[n_u++] = ((uint64_t)sc<<32)|(n_v-n_v0); } // if(n_v != (int64_t)res->n) { // fprintf(stderr, "\nERROR->n_v:%ld, res->n:%d, qlen:%ld\n", n_v, (int32_t)res->n, qlen); // for (k = 0; k < (int64_t)res->n; k++) { // fprintf(stderr, "(%ld)srt-sc:%lu, srt-i:%u\n", k, srt[k]>>32, (uint32_t)srt[k]); // } // for (k = 0; k < (int64_t)res->n; k++) { // fprintf(stderr, "(%ld)track-sc:%lu, track-pi:%lu\n", k, track[k]>>32, track[k]&((uint64_t)0x7FFFFFFF)); // } // } // if(n_v && ex->a[0].qn == 6) { // for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { // n_v0 = n_v; n_v += (uint32_t)idx[k]; // fprintf(stderr, "\n"); // for (i = n_v0; i < n_v; i++) { // fprintf(stderr, "[%u, %u]\n", ex->a[i].qs, ex->a->qe); // } // } // } for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { n_v0 = n_v; n_v += (uint32_t)idx[k]; res->a[k].qn = idx[k]>>32; res->a[k].ts = n_v0; res->a[k].te = n_v; res->a[k].qs = ex->a[n_v-1].qs; res->a[k].qe = ex->a[n_v0].qe; } res->n = n_u; return res->n; } uint64_t get_het_site(haplotype_evdience_alloc *hap, uint32_t oid) { uint64_t k, l, i, occ = 0; SnpStats *s = NULL; for (k = 1, l = 0; k <= hap->length; ++k) { if (k == hap->length || hap->list[k].overlapID != hap->list[l].overlapID) { if(hap->list[l].overlapID != oid) { l = k; continue; } for (i = l; i < k; i++) { if(hap->list[i].type!=1) continue; s = &(hap->snp_stat.a[hap->list[i].overlapSite]); if(s->score == 1 && (!(s->occ_0 < 2 || s->occ_1 < 2))) { occ++; } } l = k; } } return (occ&((uint64_t)0x3FFFFFFF)); } uint64_t update_ava_het_site(haplotype_evdience_alloc *h, uint64_t oid, uint64_t *beg, uint64_t *end, uint64_t is_srt) { uint64_t k, l, i, occ = 0, n = h->length, need_srt = 0; SnpStats *s = NULL; haplotype_evdience tt; l = beg? (*beg):0; if(end) (*end) = n; if(beg) (*beg) = n; if(l < n && h->list[l].overlapID > oid){ if(end) (*end) = l; return 0; } for (k = l + 1; k <= n; ++k) { if(h->list[l].overlapID > oid) { if(end) (*end) = l; break; } if (k == n || h->list[k].overlapID != h->list[l].overlapID) { if(h->list[l].overlapID == oid) { for (i = l; i < k; i++) { if(h->list[i].type!=1) continue; s = &(h->snp_stat.a[h->list[i].overlapSite]); if(s->score == 1 && (!(s->occ_0 < 2 || s->occ_1 < 2))) { if(l+occ != i) { tt = h->list[l+occ]; h->list[l+occ] = h->list[i]; h->list[i] = tt; } if((occ>0) && (h->list[l+occ].covlist[l+occ-1].cov)) need_srt = 1; occ++; } } if(beg) (*beg) = l; if(end) (*end) = k; break; } l = k; } } // if(oid == 160) { // fprintf(stderr, "###[M::%s] l:%lu, occ:%lu\n", __func__, l, occ); // for (k = l; k < l + occ; k++) { // fprintf(stderr, "h->list[%lu]:%u\n", k, h->list[k].cov); // } // } if(occ && is_srt && need_srt) { radix_sort_hap_ev_cov_srt(h->list+l, h->list+l+occ); } return occ; } int64_t get_chain_x(overlap_region* ot, int64_t q) { int64_t x, y, off, i, lx = -1, ly = -1; Fake_Cigar* o = &(ot->f_cigar); x = get_fake_gap_pos(o, o->length - 1); off = get_fake_gap_shift(o, o->length - 1); y = x - ot->x_pos_s + ot->y_pos_s + off; // if(ot->x_id == 98 && (q == 6681 || q == 6990)) fprintf(stderr, "o->length->%u, q->%ld, y->%ld, x->%ld\n", o->length, q, y, x); if(y == q) return x; for (i = 0; i < (int64_t)o->length; i++){ x = get_fake_gap_pos(o, i); off = get_fake_gap_shift(o, i); y = x - ot->x_pos_s + ot->y_pos_s + off; // if(ot->x_id == 98 && (q == 6681 || q == 6990)) fprintf(stderr, "+i->%ld, q->%ld, y->%ld, x->%ld\n", i, q, y, x); if(q < y) { lx = x; ly = y; break; } } if(i == 0 || i == (int64_t)o->length) { fprintf(stderr, "ERROR at %s:%d, x_id->%u, y_id->%u, q->%ld, i->%ld, yi_s->%u, yi_e->%u\n", __FILE__, __LINE__, ot->x_id, ot->y_id, q, i, ot->y_pos_s, ot->y_pos_e); exit(0); } x = get_fake_gap_pos(o, i-1); off = get_fake_gap_shift(o, i-1); y = x - ot->x_pos_s + ot->y_pos_s + off; y = (((double)(q - y))/((double)(ly - y)))*((double)(lx -x)) + x; // y = q - y + x; if(y < ot->x_pos_s) y = ot->x_pos_s; if(y > ot->x_pos_e) y = ot->x_pos_e; return y; } //[s, e] double es_win_err(overlap_region* o, int64_t winLen, int64_t s, int64_t e) { int64_t si, ei, os, k, tErr = 0, tLen = 0, minE, maxS, ov; os = (o->x_pos_s/winLen)*winLen; si = (s-os)/winLen; ei = (e-os)/winLen; for (k = si+1; k <= ei-1; k++) { tLen += o->w_list[k].x_end+1-o->w_list[k].x_start; if(o->w_list[k].y_end != -1) { tErr += o->w_list[k].error; } else { tErr += o->w_list[k].x_end+1-o->w_list[k].x_start; } } k = si; maxS = MAX(s, (int64_t)(o->w_list[k].x_start)); minE = MIN(e, (int64_t)(o->w_list[k].x_end)) + 1; ov = minE > maxS? minE - maxS:0; if(ov == 0) { fprintf(stderr, "WARNNING-1, o->w_list_length->%u, o->x_id->%u, s->%ld, e->%ld, w_list_s->%lu, w_list_e->%lu, winLen->%ld, o->x_pos_s->%u, o->x_pos_e->%u, si->%ld, flag->%d\n", o->w_list_length, o->x_id, s, e, o->w_list[k].x_start, o->w_list[k].x_end, winLen, o->x_pos_s, o->x_pos_e, si, o->w_list[k].y_end); } tLen += ov/**o->w_list[k].x_end+1-o->w_list[k].x_start**/; if(o->w_list[k].y_end != -1) { tErr += (ov*o->w_list[k].error)/(o->w_list[k].x_end+1-o->w_list[k].x_start); } else { tErr += ov/**o->w_list[k].x_end+1-o->w_list[k].x_start**/; } k = ei; maxS = MAX(s, (int64_t)(o->w_list[k].x_start)); minE = MIN(e, (int64_t)(o->w_list[k].x_end)) + 1; ov = minE > maxS? minE - maxS:0; if(ov == 0) { fprintf(stderr, "WARNNING-2, o->w_list_length->%u, o->x_id->%u, s->%ld, e->%ld, w_list_s->%lu, w_list_e->%lu, winLen->%ld, o->x_pos_s->%u, o->x_pos_e->%u, ei->%ld, flag->%d\n", o->w_list_length, o->x_id, s, e, o->w_list[k].x_start, o->w_list[k].x_end, winLen, o->x_pos_s, o->x_pos_e, ei, o->w_list[k].y_end); } tLen += ov/**o->w_list[k].x_end+1-o->w_list[k].x_start**/; if(o->w_list[k].y_end != -1) { tErr += (ov*o->w_list[k].error)/(o->w_list[k].x_end+1-o->w_list[k].x_start); } else { tErr += ov/**o->w_list[k].x_end+1-o->w_list[k].x_start**/; } return ((double)tErr)/((double)tLen); } int64_t gen_contain_chain(const ul_idx_t *uref, utg_ct_t *p, overlap_region* o, kv_ul_ov_t *chains, double diff_ec_ul, int64_t winLen, void *km) { int64_t y_s, y_e, y_bs, y_be, x_s, x_e, q_s, q_e; if(o->y_pos_strand) { y_s = uref->ug->u.a[o->y_id].len - p->e; y_e = uref->ug->u.a[o->y_id].len - p->s - 1; } else { y_s = p->s; y_e = p->e - 1; } y_s = MAX(y_s, (int64_t)o->y_pos_s); y_e = MIN(y_e, (int64_t)o->y_pos_e); if(y_s > y_e) return 0; x_s = get_chain_x(o, y_s); x_e = get_chain_x(o, y_e) + 1; if(x_s >= x_e) fprintf(stderr, "+++y_s->%ld, y_e->%ld, x_s->%ld, x_e->%ld\n", y_s, y_e, x_s, x_e); if(o->y_pos_strand) { y_bs = uref->ug->u.a[o->y_id].len - (y_e+1); y_be = uref->ug->u.a[o->y_id].len - y_s; } else { y_bs = y_s; y_be = y_e + 1; } q_s = 0; q_e = p->e - p->s; if(p->x&1) { q_s += (p->e - y_be); q_e -= (y_bs - p->s); } else { q_s += (y_bs - p->s); q_e -= (p->e - y_be); } // if(q_s < 0 || q_e < 0 || q_s >= (int64_t)(p->e - p->s) || q_e > (int64_t)(p->e - p->s)) fprintf(stderr, "ERROR\n"); if(winLen > 0 && diff_ec_ul > 0 && es_win_err(o, winLen, x_s, x_e-1) > diff_ec_ul) return 0; ul_ov_t *x = NULL; kv_pushp_km(km, ul_ov_t, *chains, &x); x->qn = o->x_id; x->qs = x_s; x->qe = x_e; /**x->tn = p->x>>1;**/x->tn = (uint32_t)(0x80000000); x->tn |= (p->x>>1); x->ts = q_s; x->te = q_e; x->el = 1;x->sec = 0; x->rev = ((o->y_pos_strand == (p->x&1))?0:1); // if(x->qn == 0 /**&& ((x->tn<<1)>>1) == 302**/) { // /**if(o->x_id == 0 && (o->y_id == 46 || o->y_id == 48))**/ { // // fprintf(stderr, "\nUL[%u]\t%u\t%u\t%c\tUTG[%u]\t%u\t%u\n", o->x_id, o->x_pos_s, o->x_pos_e, // // "+-"[o->y_pos_strand], o->y_id, o->y_pos_s, o->y_pos_e); // // fprintf(stderr, "Contain[%u]\t%c\ts[%u]\te[%u]\n", p->x>>1, "+-"[p->x&1], p->s, p->e); // fprintf(stderr, "U[%u]\t%u\t%u\t%c\tR[%u]\t%u\t%u\tUid[%u]\n", x->qn, x->qs, x->qe, // "+-"[x->rev], ((x->tn<<1)>>1), x->ts, x->te, o->y_id); // } // } return 1; } int64_t debug_utg_ct_t(const ul_idx_t *uref, overlap_region* o, utg_ct_t *ct_a, int64_t ct_n, ma_utg_t *u, utg_ct_t *z, haplotype_evdience *he_a, int64_t he_n) { int64_t k, i, l, rs, re, ss, m = 0; utg_ct_t *p = NULL; if(ct_a && ct_n) { for (i = 0; i < ct_n; i++) { p = &(ct_a[i]); for (k = 0; k < he_n; k++) { ss = o->y_pos_strand?uref->ug->u.a[o->y_id].len - he_a[k].cov - 1:he_a[k].cov; if(ss >= p->s && ss < p->e) break; } if(k < he_n) m++; } } if(u) { for (i = l = 0; i < u->n; i++) { rs = l; re = l + Get_READ_LENGTH(R_INF, (u->a[i]>>33)); l += (uint32_t)u->a[i]; for (k = 0; k < he_n; k++) { ss = o->y_pos_strand?uref->ug->u.a[o->y_id].len - he_a[k].cov - 1:he_a[k].cov; if(ss >= rs && ss < re) break; } if(k < he_n) m++; } } if(z) { for (k = 0; k < he_n; k++) { ss = o->y_pos_strand?uref->ug->u.a[o->y_id].len - he_a[k].cov - 1:he_a[k].cov; if(ss >= z->s && ss < z->e) break; } if(k < he_n) m++; } return m; } int64_t rescue_contain_ul_chains(const ul_idx_t *uref, overlap_region* o, haplotype_evdience *he_a, int64_t he_n, utg_ct_t *ct_a, int64_t ct_n, kv_ul_ov_t *chains, double diff_ec_ul, int64_t winLen, int64_t rescue_trans, void *km) { int64_t i, k, ss, ff, t0 = 0; uint64_t ys, ye; utg_ct_t *p = NULL; // if(o->x_id == 0) { // fprintf(stderr, "\no->y_id->%u\n", o->y_id); // for (i = 0; i < ct_n; i++) { // p = &(ct_a[i]); // fprintf(stderr, "***rid->%u, rev->%u, s->%u, e->%u\n", p->x>>1, p->x&1, p->s, p->e); // } // } if(o->y_pos_strand == 0){ ys = o->y_pos_s; ye = o->y_pos_e + 1; for (i = k = 0; i < ct_n; i++) { p = &(ct_a[i]); if(p->e <= ys) continue; if(p->s >= ye) break; ff = 1; if(he_a && he_n > 0) { for (; k < he_n; k++) { if(he_a[k].cov >= p->s && he_a[k].cov < p->e) { ff = 0; break; } if(he_a[k].cov >= p->e) break; } } // if(ff == debug_utg_ct_t(uref, o, p, he_a, he_n)) fprintf(stderr, "ERROR\n"); if(ff) { ///push ovlp t0 += gen_contain_chain(uref, p, o, chains, diff_ec_ul, winLen, km); } else if(rescue_trans) { if(gen_contain_chain(uref, p, o, chains, diff_ec_ul, winLen, km)){ t0++; chains->a[chains->n-1].el = 0; } } // if(!ff) t0++; } } else { ys = uref->ug->u.a[o->y_id].len - (o->y_pos_e+1); ye = uref->ug->u.a[o->y_id].len - o->y_pos_s; for (i = 0, k = he_n - 1; i < ct_n; i++) { p = &(ct_a[i]); if(p->e <= ys) continue; if(p->s >= ye) break; ff = 1; if(he_a && he_n > 0) { for (; k >= 0; k--) { ss = uref->ug->u.a[o->y_id].len - he_a[k].cov - 1; if(ss >= p->s && ss < p->e) { ff = 0; break; } if(ss >= p->e) break; } } // if(ff == debug_utg_ct_t(uref, o, p, he_a, he_n)) fprintf(stderr, "ERROR\n"); if(ff) { ///push ovlp t0 += gen_contain_chain(uref, p, o, chains, diff_ec_ul, winLen, km); } else if(rescue_trans) { if(gen_contain_chain(uref, p, o, chains, diff_ec_ul, winLen, km)){ t0++; chains->a[chains->n-1].el = 0; } } // if(!ff) t0++; } } // if(debug_utg_ct_t(uref, o, ct_a, ct_n, he_a, he_n)!=t0) fprintf(stderr, "ERROR\n"); // fprintf(stderr, "***[M::%s] o->y_id:%u, chains->n:%u\n", __func__, o->y_id, (uint32_t)chains->n); return t0; } int64_t rescue_trans_ul_chains(const ul_idx_t *uref, overlap_region* o, haplotype_evdience *he_a, int64_t he_n, ma_utg_t *u, kv_ul_ov_t *chains, double diff_ec_ul, int64_t winLen, int64_t rescue_trans, uint64_t *cis_occ, void *km) { uint64_t ys, ye, i, l; int64_t k, ff, ss, t0 = 0; utg_ct_t p; if(cis_occ) (*cis_occ) = 0; if(o->y_pos_strand == 0) { ys = o->y_pos_s; ye = o->y_pos_e + 1; for (i = k = l = 0; i < u->n; i++) { p.x = u->a[i]>>32; p.s = l; p.e = l + Get_READ_LENGTH(R_INF, (u->a[i]>>33)); l += (uint32_t)u->a[i]; if(p.e <= ys) continue; if(p.s >= ye) break; ff = 1; if(he_a && he_n) { for (; k < he_n; k++) { if(he_a[k].cov >= p.s && he_a[k].cov < p.e) { ff = 0; break; } if(he_a[k].cov >= p.e) break; } } // if(ff == debug_utg_ct_t(uref, o, 0, 0, 0, &p, he_a, he_n)) fprintf(stderr, "ERROR\n"); if(ff) { ///push ovlp t0 += gen_contain_chain(uref, &p, o, chains, diff_ec_ul, winLen, km); } else if(rescue_trans) { if(gen_contain_chain(uref, &p, o, chains, diff_ec_ul, winLen, km)){ t0++; chains->a[chains->n-1].el = 0; if(cis_occ) (*cis_occ)++; } } // if(!ff) t0++; } } else { ys = uref->ug->u.a[o->y_id].len - (o->y_pos_e+1); ye = uref->ug->u.a[o->y_id].len - o->y_pos_s; for (i = l = 0, k = he_n - 1; i < u->n; i++) { p.x = u->a[i]>>32; p.s = l; p.e = l + Get_READ_LENGTH(R_INF, (u->a[i]>>33)); l += (uint32_t)u->a[i]; if(p.e <= ys) continue; if(p.s >= ye) break; ff = 1; if(he_a && he_n) { for (; k >= 0; k--) { ss = uref->ug->u.a[o->y_id].len - he_a[k].cov - 1; if(ss >= p.s && ss < p.e) { ff = 0; break; } if(ss >= p.e) break; } } // if(ff == debug_utg_ct_t(uref, o, 0, 0, 0, &p, he_a, he_n)) fprintf(stderr, "ERROR\n"); if(ff) { ///push ovlp t0 += gen_contain_chain(uref, &p, o, chains, diff_ec_ul, winLen, km); } else if(rescue_trans) { if(gen_contain_chain(uref, &p, o, chains, diff_ec_ul, winLen, km)){ t0++; chains->a[chains->n-1].el = 0; if(cis_occ) (*cis_occ)++; } } // if(!ff) t0++; } } // if(debug_utg_ct_t(uref, o, NULL, 0, u, he_a, he_n)!=t0) fprintf(stderr, "ERROR\n"); // fprintf(stderr, "t0->%ld\n", t0); return t0; } int64_t dedup_sort_ul_ov_t(ul_ov_t *a, int64_t a_n) { int64_t k, l, z, r, i, qo, to; float rr = 0.9; for (k = 1, l = i = 0; k <= a_n; k++) { if(k == a_n || a[k].tn != a[l].tn) {///remove the duplicated contained alignments for (z = l; z < k; z++) { for (r = i-1; r >= 0 && a[r].tn == a[z].tn; r--){ /** if(a[z].qn == a[r].qn && a[z].qs == a[r].qs && a[z].qe == a[r].qe && a[z].tn == a[r].tn && a[z].ts == a[r].ts && a[z].te == a[r].te && a[z].sec == a[r].sec && a[z].el == a[r].el && a[z].rev == a[r].rev) { break; } **/ if(a[z].qn == a[r].qn && a[z].tn == a[r].tn && a[z].rev == a[r].rev) { qo = ((MIN(a[z].qe, a[r].qe) > MAX(a[z].qs, a[r].qs))? MIN(a[z].qe, a[r].qe) - MAX(a[z].qs, a[r].qs):0); to = ((MIN(a[z].te, a[r].te) > MAX(a[z].ts, a[r].ts))? MIN(a[z].te, a[r].te) - MAX(a[z].ts, a[r].ts):0); if(qo >= ((a[r].qe - a[r].qs)*rr) && qo >= ((a[z].qe - a[z].qs)*rr) && to >= ((a[r].te - a[r].ts)*rr) && to >= ((a[z].te - a[z].ts)*rr)) { break; } } } if(r >= 0 && a[r].tn == a[z].tn) { if(a[z].el) a[r].el = 1; continue; } a[i++] = a[z]; } l = k; } } return i; } uint32_t check_contain_pair(const ug_opt_t *uopt, uint32_t x, uint32_t y, uint32_t check_el) { ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang; uint64_t z, qn, tn; int32_t r = 1; asg_arc_t e; for (z = 0; z < src[x].length; z++) { if(check_el && (!src[x].buffer[z].el)) continue; qn = Get_qn(src[x].buffer[z]); tn = Get_tn(src[x].buffer[z]); if(tn != y) continue; r = ma_hit2arc(&(src[x].buffer[z]), Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &e); if(r == MA_HT_QCONT || r == MA_HT_TCONT) break; } if(z < src[x].length) return 1; return 0; } void debug_contain_ovlps(ul_ov_t *a, uint64_t a_n, const ug_opt_t *uopt) { uint64_t k, i, f; ul_ov_t *z = NULL, *w = NULL; for (k = 0; k < a_n; k++) { z = &(a[k]); if(!(z->tn&((uint32_t)(0x80000000)))) continue; for (i = 0, f = z->qn; i < a_n; i++) { w = &(a[i]); if(i == k) continue; if(w->tn&((uint32_t)(0x80000000))) continue; if(z->qs >= w->qs && z->qe <= w->qe && check_contain_pair(uopt, (z->tn<<1)>>1, w->tn, 1)) { f = (uint32_t)-1; break; } } if(z->qn != f) fprintf(stderr, "ERROR\n"); } } ma_hit_t* query_ovlp_src(const ug_opt_t *uopt, uint32_t v, uint32_t w, int64_t o, double diff_ec_ul, uint32_t *ol) { ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang, d, l, max_l; uint64_t z, qn, tn, x = v>>1; int32_t r = 1; asg_arc_t e; l = (o>=0?o:-o); //l *= diff_ec_ul; if(l <= 0) return NULL; for (z = 0; z < src[x].length; z++) { qn = Get_qn(src[x].buffer[z]); tn = Get_tn(src[x].buffer[z]); if(tn != (w>>1)) continue; r = ma_hit2arc(&(src[x].buffer[z]), Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &e); if(r < 0) continue; if((e.ul>>32) != v || e.v != w) continue; // if(v == 56 && w == 25) fprintf(stderr, "+xxxx, o:%ld, e.ol:%u\n", o, e.ol); // if(v == 25 && w == 56) fprintf(stderr, "-xxxx, o:%ld, e.ol:%u\n", o, e.ol); d = (o>=e.ol?o-e.ol:e.ol-o); max_l = MAX(l, e.ol); if(d <= (max_l*diff_ec_ul)) { if(ol) (*ol) = e.ol; return &(src[x].buffer[z]); } } return NULL; } int64_t infer_rovlp(ul_ov_t *li, ul_ov_t *lj, uc_block_t *bi, uc_block_t *bj, All_reads *ridx, ma_ug_t *ug) { int64_t in, is, ie, irev, iqs, iqe, jn, js, je, jrev, jqs, jqe, ir, jr, ts, te, max_s, min_e, s_shift, e_shift; if(li) { in = ug?ug->u.a[li->tn].len:Get_READ_LENGTH(R_INF, li->tn); is = li->ts; ie = li->te; irev = li->rev; iqs = li->qs; iqe = li->qe; } else if(bi) { in = ug?ug->u.a[bi->hid].len:Get_READ_LENGTH(R_INF, bi->hid); is = bi->ts; ie = bi->te; irev = bi->rev; iqs = bi->qs; iqe = bi->qe; } else { return 0; } if(lj) { jn = ug?ug->u.a[lj->tn].len:Get_READ_LENGTH(R_INF, lj->tn); js = lj->ts; je = lj->te; jrev = lj->rev; jqs = lj->qs; jqe = lj->qe; } else if(bj) { jn = ug?ug->u.a[bj->hid].len:Get_READ_LENGTH(R_INF, bj->hid); js = bj->ts; je = bj->te; jrev = bj->rev; jqs = bj->qs; jqe = bj->qe; } else { return 0; } max_s = MAX(iqs, jqs); min_e = MIN(iqe, jqe); if(min_e <= max_s) return 0; s_shift = get_offset_adjust(max_s - iqs, iqe-iqs, ie-is); e_shift = get_offset_adjust(iqe - min_e, iqe-iqs, ie-is); if(irev) { ts = s_shift; s_shift = e_shift; e_shift = ts; } is += s_shift; ie-= e_shift; // if(li && lj && li->tn == 324 && lj->tn == 319 && li->qs == 63841) { // fprintf(stderr, "+++in:%ld, is:%ld, ie:%ld, irev:%ld, jn:%ld, js:%ld, je:%ld, jrev:%ld\n", in, is, ie, irev, jn, js, je, jrev); // } s_shift = get_offset_adjust(max_s - jqs, jqe-jqs, je-js); e_shift = get_offset_adjust(jqe - min_e, jqe-jqs, je-js); if(jrev) { ts = s_shift; s_shift = e_shift; e_shift = ts; } js += s_shift; je-= e_shift; if(irev) { ts = in - ie; te = in - is; is = ts; ie = te; } if(jrev) { ts = jn - je; te = jn - js; js = ts; je = te; } // if(li && lj && li->tn == 324 && lj->tn == 319 && li->qs == 63841) { // fprintf(stderr, "---in:%ld, is:%ld, ie:%ld, irev:%ld, jn:%ld, js:%ld, je:%ld, jrev:%ld\n", in, is, ie, irev, jn, js, je, jrev); // } if(is <= js) { js -= is; is = 0; } else { is -= js; js = 0; } ir = in - ie; jr = jn - je; if(ir <= jr){ ie = in; je += ir; } else { je = jn; ie += jr; } ir = ie - is; jr = je - js; return MAX(ir, jr); } void debug_infer_read_ovlp(const ug_opt_t *uopt, double diff_ec_ul, ul_ov_t *li, ul_ov_t *lj, ma_utg_t *u, uint32_t i_idx, uint32_t j_idx, All_reads *ridx, ma_ug_t *ug) { uint32_t li_v, lj_v; ma_hit_t *t = NULL; li_v = (((uint32_t)(li->tn))<<1)|((uint32_t)(li->rev)); lj_v = (((uint32_t)(lj->tn))<<1)|((uint32_t)(lj->rev)); if(lj->qe <= li->qs || li_v == lj_v) fprintf(stderr, "ERROR-1\n"); t = query_ovlp_src(uopt, li_v^1, lj_v^1, infer_rovlp(li, lj, NULL, NULL, ridx, ug), diff_ec_ul, NULL); // ((int64_t)(lj->qe))-((int64_t)(li->qs)) if(!t /**&& (li_v^1) == 648 && (lj_v^1) == 638 && li->qs == 63841**/) { fprintf(stderr, "ERROR-2, li_v^1->%u, li->qs->%u, li->qe->%u, lj_v^1->%u, lj->qs->%u, lj->qe->%u, infer_rovlp->%ld\n", li_v^1, li->qs, li->qe, lj_v^1, lj->qs, lj->qe, infer_rovlp(li, lj, NULL, NULL, ridx, ug)); } } uint64_t infer_read_ovlp(const ul_idx_t *uref, overlap_region_alloc* olist, kv_ul_ov_t *in, kv_ul_ov_t *res, double diff_ec_ul, int64_t winLen, const ug_opt_t *uopt, ul_contain *ct, void *km) { uint64_t t, k, l, t_0, pb, cut = res->n, c_occ = 0;; ma_ug_t *ug = uref->ug; ma_utg_t *u = NULL; overlap_region* o = NULL; ul_ov_t *z = NULL; utg_ct_t p; // res->n = 0; for (t = 0; t < in->n; t++) { if(!(in->a[t].tn&(uint32_t)(0x80000000))) {///uid u = &(ug->u.a[in->a[t].tn]); o = &(olist->list[in->a[t].qn]); assert(o->y_id == in->a[t].tn); for (k = l = 0, pb = res->n+2; k < u->n; k++) { p.x = u->a[k]>>32; p.s = l; p.e = l + Get_READ_LENGTH(R_INF, (u->a[k]>>33)); l += (uint32_t)u->a[k]; if(p.e <= in->a[t].ts) continue; if(p.s >= in->a[t].te) break; t_0 = gen_contain_chain(uref, &p, o, res, -1, -1, km); assert(t_0 > 0); // if(t_0 == 0) { // fprintf(stderr, "ERROR-2, o->x_id:%u, o->y_id:%u, k:%lu, u->n:%lu, p.s:%u, p.e:%u, ts:%u, te:%u, rev:%u\n", // o->x_id, o->y_id, k, (uint64_t)u->n, p.s, p.e, in->a[t].ts, in->a[t].te, in->a[t].rev); // } res->a[res->n-1].el = in->a[t].el; res->a[res->n-1].sec = in->a[t].sec; res->a[res->n-1].tn <<= 1; res->a[res->n-1].tn >>= 1; res->a[res->n-1].qn = o->x_id; if(res->n >= pb) { if(in->a[t].rev == 0) { // if(res->a[res->n-1].qs > res->a[res->n-2].qe) fprintf(stderr, "ERROR-3\n"); if(!(res->a[res->n-2].qs<=res->a[res->n-1].qs && res->a[res->n-1].qs <= res->a[res->n-2].qe && res->a[res->n-2].qe <= res->a[res->n-1].qe)) { fprintf(stderr, "ERROR-3\n"); } // if(res->a[res->n-1].qs == res->a[res->n-2].qe) { // if(res->a[res->n-2].qe < in->a[t].qe) res->a[res->n-2].qe++; // else if(res->a[res->n-1].qs > 0) res->a[res->n-1].qs--; // } } else { if(!(res->a[res->n-1].qs<=res->a[res->n-2].qs && res->a[res->n-2].qs <= res->a[res->n-1].qe && res->a[res->n-1].qe <= res->a[res->n-2].qe)) { fprintf(stderr, "ERROR-4\n"); } } // debug_infer_read_ovlp(uopt, diff_ec_ul, // in->a[t].rev?&(res->a[res->n-2]):&(res->a[res->n-1]), // in->a[t].rev?&(res->a[res->n-1]):&(res->a[res->n-2]), u, k, k-1); } } } else {///rid kv_push(ul_ov_t, *res, in->a[t]); // res->a[res->n-1].tn <<= 1; // res->a[res->n-1].tn >>= 1; res->a[res->n-1].qn = o->x_id; c_occ++; } } if(res->n != cut) { radix_sort_ul_ov_srt_qe(res->a + cut, res->a + res->n); if(c_occ) { int64_t ci, cn = cut; for (k = cut; k < res->n; k++) { z = &(res->a[k]); if(z->tn&((uint32_t)(0x80000000))) continue; if(ct->is_c.a[z->tn] == 0) continue; for (ci = k+1; ci < (int64_t)(res->n); ci++) { if(res->a[ci].qe > z->qe) break; if(res->a[ci].qn == (uint32_t)-1) continue; if(!(res->a[ci].tn&((uint32_t)(0x80000000)))) continue; if(z->qs <= res->a[ci].qs && z->qe >= res->a[ci].qe) { if(check_contain_pair(uopt, (res->a[ci].tn<<1)>>1, z->tn, 1)) { res->a[ci].qn = (uint32_t)-1; } } } for (ci = k-1; ci >= cn; ci--) { if(res->a[ci].qe <= z->qs) break; if(res->a[ci].qn == (uint32_t)-1) continue; if(!(res->a[ci].tn&((uint32_t)(0x80000000)))) continue; if(z->qs <= res->a[ci].qs && z->qe >= res->a[ci].qe) { if(check_contain_pair(uopt, (res->a[ci].tn<<1)>>1, z->tn, 1)) { res->a[ci].qn = (uint32_t)-1; } } } } // debug_contain_ovlps(res->a+cut, res->n-cut, uopt); for (k = l = cut; k < res->n; k++) { if(res->a[k].qn == (uint32_t)-1) continue; if(k != l) { res->a[l] = res->a[k]; } res->a[l].tn <<= 1; res->a[l].tn >>= 1; ++l; } res->n = l; } } return res->n - cut; } int64_t gl_chain_refine(overlap_region_alloc* olist, Correct_dumy* dumy, haplotype_evdience_alloc *hap, glchain_t *ll, const ul_idx_t *uref, double diff_ec_ul, int64_t winLen, int64_t qlen, const ug_opt_t *uopt, void *km) { ll->tk.n = ll->lo.n = 0; kv_ul_ov_t *idx = &(ll->lo); ul_contain *ct = uref->ct; gl_chain_gen(olist, uref, idx, 0, km); if(idx->n == 0) return 0; kv_resize_km(km, ul_ov_t, ll->tk, idx->n); kv_resize_km(km, uint64_t, ll->srt.a, idx->n); kv_resize_km(km, uint64_t, hap->snp_srt, idx->n); if(gl_exact_chain(idx, &(ll->tk), uref, G_CHAIN_BW, diff_ec_ul, qlen, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, km)) { kv_ul_ov_t *chains = &(ll->tk); ul_ov_t *p = NULL; uint64_t k, z, ff, s, e, an, cn, sft = 50, si = 0, ei = 0, resc = 0, chains_pl = chains->n; radix_sort_ul_ov_srt_qs(idx->a, idx->a + idx->n); for (k = 0; k < olist->length; k++) { if(olist->list[k].is_match!=2) continue; s = olist->list[k].x_pos_s; e = olist->list[k].x_pos_e+1; for (z = ff = 0; z < idx->n; z++) { if((s+sft) >= idx->a[z].qs && e <= (idx->a[z].qe+sft)) { ff = 1; break; } if(idx->a[z].qs >= (e+sft)) break; } cn = ((uint32_t)(ct->idx.a[olist->list[k].y_id])); if(ff && cn==0) continue; an = update_ava_het_site(hap, k, &si, &ei, cn); // if(an != get_het_site(hap, k)) fprintf(stderr, "an->%lu, get_het_site->%lu\n", an, get_het_site(hap, k)); if(cn > 0 && an > 0) { resc += rescue_contain_ul_chains(uref, &(olist->list[k]), hap->list+si, an, ct->rids.a + ((ct->idx.a[olist->list[k].y_id])>>32), cn, chains, diff_ec_ul, winLen, 0, km); } if(ff == 0) { kv_pushp_km(km, ul_ov_t, *chains, &p); p->qn = k/**olist->list[k].x_id**/; p->qs = olist->list[k].x_pos_s; p->qe = olist->list[k].x_pos_e+1; p->tn = olist->list[k].y_id; p->el = 0; p->sec = (an&((uint64_t)0x3FFFFFFF))/**get_het_site(hap, k)**/; p->rev = olist->list[k].y_pos_strand; if(p->rev) { p->ts = uref->ug->u.a[p->tn].len - (olist->list[k].y_pos_e+1); p->te = uref->ug->u.a[p->tn].len - olist->list[k].y_pos_s; } else { p->ts = olist->list[k].y_pos_s; p->te = olist->list[k].y_pos_e+1; } } si = ei; } if(resc > 0) { radix_sort_ul_ov_srt_tn(chains->a + chains_pl, chains->a + chains->n); ff = dedup_sort_ul_ov_t(chains->a + chains->n - resc, resc); chains->n = chains->n - resc + ff; } } if(ll->tk.n > 0) infer_read_ovlp(uref, olist, &(ll->tk), &(ll->lo), diff_ec_ul, winLen, uopt, ct, km); else ll->lo.n = 0; return 1; } /** void fill_edge_weight(ul_ov_t *a, int64_t a_n, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen) { uint32_t li_v, lj_v; ul_ov_t *li = NULL, *lj = NULL; int64_t mm_ovlp, x, i, j, o; ma_hit_t *t = NULL; for (i = 0; i < a_n; i++) { li = &(a[i]); li_v = (li->tn<<1)|li->rev; mm_ovlp = max_ovlp_src(uopt, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, a, x); for (j = x; j >= 0; --j) { // collect potential destination vertices lj = &(a[j]); lj_v = (lj->tn<<1)|lj->rev; if(lj->qe <= li->qs) break; if(li_v == lj_v) continue; t = query_ovlp_src(uopt, li_v, lj_v, ((int64_t)(lj->qe))-((int64_t)(li->qs)), diff_ec_ul); if(t) { t->bl; } } } } **/ int64_t get_ecov_adv_back(const ul_idx_t *uref, const ug_opt_t *uopt, uint32_t v, uint32_t w, int64_t bw, double diff_ec_ul, int64_t dq, uint32_t *is_contain) { int64_t dt = -1, dif, mm; if(is_contain) (*is_contain) = 0; const asg_t *g = uref?uref->ug->g:NULL; uint32_t nv, i; asg_arc_t *av = NULL; if(g) { nv = asg_arc_n(g, v); av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].del || av[i].v != w) continue; dt = av[i].ol; break; } } if(dt < 0 && uopt) { ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang; uint64_t z, qn, tn, x = v>>1; int32_t r = 1; asg_arc_t e; for (z = 0; z < src[x].length; z++) { qn = Get_qn(src[x].buffer[z]); tn = Get_tn(src[x].buffer[z]); if(tn != (w>>1)) continue; r = ma_hit2arc(&(src[x].buffer[z]), Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &e); if(r < 0) { if(r == MA_HT_QCONT || r == MA_HT_TCONT) { if(src[x].buffer[z].rev == ((uint32_t)(v^w))) { dt = Get_qe(src[x].buffer[z]) - Get_qs(src[x].buffer[z]); if(dt < Get_te(src[x].buffer[z]) - Get_ts(src[x].buffer[z])) { dt = Get_te(src[x].buffer[z]) - Get_ts(src[x].buffer[z]); } if(is_contain) (*is_contain) = 1; break; } } continue; } if((e.ul>>32) != v || e.v != w) continue; dt = e.ol; break; } } if(dt < 0) return 0; dif = (dq>dt? dq-dt:dt-dq); mm = MAX(dq, dt); mm *= diff_ec_ul; if(mm < bw) mm = bw; // if((v>>1) == 1163 && (w>>1) == 1168) fprintf(stderr, ">>>>>>dis_q:%ld, dis_t:%ld, dif:%ld, mm:%ld\n", dis_q, dis_t, dif, mm); if(dif <= mm) return 1; return 0; } ma_hit_t *get_ug_edge_src(ma_ug_t *ug, ma_hit_t_alloc *src, int64_t max_hang, int64_t min_ovlp, uint32_t uv, uint32_t uw) { if(ug->u.a[uv>>1].circ || ug->u.a[uw>>1].circ) return NULL; uint32_t v, w, k, qn, tn; int32_t r; asg_arc_t t; v = ((uv&1)?(ug->u.a[uv>>1].start^1):(ug->u.a[uv>>1].end^1)); w = ((uw&1)?(ug->u.a[uw>>1].end):(ug->u.a[uw>>1].start)); ma_hit_t_alloc *x = &(src[v>>1]); for (k = 0; k < x->length; k++) { qn = Get_qn(x->buffer[k]); tn = Get_tn(x->buffer[k]); if(qn == (v>>1) && tn == (w>>1)) { r = ma_hit2arc(&(x->buffer[k]), Get_READ_LENGTH(R_INF, v>>1), Get_READ_LENGTH(R_INF, w>>1), max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0) continue; if((t.ul>>32)!=v || t.v!=w) continue; return &(x->buffer[k]); } } return NULL; } ///mode: 0->ug; 1->read int64_t get_ecov_adv(const ul_idx_t *uref, const ug_opt_t *uopt, uint32_t v, uint32_t w, int64_t bw, double diff_ec_ul, int64_t dq, uint64_t mode, int64_t *contain_off) { int64_t dt = -1, dif, mm; (*contain_off) = 0; uint32_t nv, i; asg_arc_t *av = NULL; ma_hit_t *x = NULL; if(!mode) { const asg_t *g = uref?uref->ug->g:NULL; nv = asg_arc_n(g, v); av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].del || av[i].v != w) continue; dt = av[i].ol; (*contain_off) = av[i].ou; // if(v==1772 && w==1769) fprintf(stderr, "+++v:%u, w:%u, ou:%u\n", v, w, av[i].ou); // if((v>>1) == 3012 && (w>>1) == 3011) fprintf(stderr, "******************\n"); if(av[i].ou >= OU_MASK) { x = get_ug_edge_src(uref->ug, uopt->sources, uopt->max_hang, uopt->min_ovlp, av[i].ul>>32, av[i].v); (*contain_off) = x->cc; // if(v==1772 && w==1769) fprintf(stderr, "---v:%u, w:%u, cc:%u\n", v, w, x->cc); } break; } }else { ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang; uint64_t z, qn, tn, x = v>>1; int32_t r = 1; asg_arc_t e; for (z = 0; z < src[x].length; z++) { qn = Get_qn(src[x].buffer[z]); tn = Get_tn(src[x].buffer[z]); if(tn != (w>>1)) continue; r = ma_hit2arc(&(src[x].buffer[z]), Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &e); if(r < 0) continue; if((e.ul>>32) != v || e.v != w) continue; dt = e.ol; (*contain_off) = src[x].buffer[z].cc; break; } } if(dt < 0) return 0; dif = (dq>dt? dq-dt:dt-dq); mm = MAX(dq, dt); mm *= diff_ec_ul; if(mm < bw) mm = bw; // if((v>>1) == 1163 && (w>>1) == 1168) fprintf(stderr, ">>>>>>dis_q:%ld, dis_t:%ld, dif:%ld, mm:%ld\n", dis_q, dis_t, dif, mm); if(dif <= mm) return 1; return 0; } void get_rr_tse(const ul_idx_t *uref, ul_ov_t *li, uint32_t *ts, uint32_t *te, uint32_t *tl) { (*tl) = uref?uref->ug->g->seq[li->tn].len:Get_READ_LENGTH(R_INF, li->tn); if(!(li->rev)) { (*ts) = li->ts; (*te) = li->te; } else { (*ts) = (*tl) - li->te; (*te) = (*tl) - li->ts; } } /** uint32_t checkM(uint32_t v, uint32_t l, const ul_idx_t *uref, const asg_t *g, uint32_t in, uint32_t its, uint32_t iqs, uint32_t iqe, int64_t bw, double diff_ec_ul, int64_t qlen, ul_ov_t *a, uint32_t a_n) { int64_t vl = uref?uref->ug->g->seq[v>>1].len:Get_READ_LENGTH(R_INF, (v>>1)), t_dis, q_dis, kcs, mm_ovlp, x, k; t_dis = ((int64_t)(l + vl)) - ((int64_t)(in - its)); kcs = iqs; kcs -= t_dis; if(kcs < 0) kcs = 0; uint32_t nv = asg_arc_n(g, v), i, lk_v, kts, kte, kn; asg_arc_t *av = asg_arc_a(g, v), *p = NULL; mm_ovlp = -1; ul_ov_t *lk; for (i = 0, p = NULL; i < nv; i++) { if(av[i].del) continue; if((int32_t)(av[i].ol) > mm_ovlp) { p = &(av[i]); mm_ovlp = av[i].ol; } } if(!p) return 0; x = (kcs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += kcs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(a_n, a, x); for (k = x; k >= 0; --k) { lk = &(a[k]); lk_v = ((lk->tn<<1)|lk->rev)^1; if(lk->qe <= kcs) break;//evan this pair has a overlap, its length will be very small; just ignore if(lk->qs >= kcs) continue; // lk is contained in li on the query coordinate get_rr_tse(uref, lk, &kts, &kte, &kn); ///t_dis and q_dis might be < 0 t_dis = ((int64_t)(l + kn - kte)) - ((int64_t)(in - its)); q_dis = ((int64_t)(iqs)) - ((int64_t)(lk->qe)); } } void best_path_ext(const ul_idx_t *uref, const ug_opt_t *uopt, int64_t g_gap, ul_ov_t *a, uint32_t a_n, int64_t bw, double diff_ec_ul, uint64_t *track, ul_ov_t *li) { if(a_n <= 0) return; const asg_t *g = uref?uref->ug->g:NULL; asg_arc_t *av = NULL, *p = NULL; ul_ov_t *lk; uint32_t nv, i, v, io, in, its, ite, kn, kts, kte; int64_t mm, l, max_dist, k, t_dis, q_dis; get_rr_tse(uref, li, &its, &ite, &in); io = in - ite; max_dist = g_gap + in - its; if(g) { v = (((li->tn<<1)|li->rev)^1); mm = 1; l = 0; while (mm >= 0) { nv = asg_arc_n(g, v); av = asg_arc_a(g, v); mm = -1; for (i = 0, p = NULL; i < nv; i++) { if(av[i].del) continue; if((int32_t)(av[i].ol) > mm) { p = &(av[i]); mm = av[i].ol; } } if(p) { l += (uint32_t)(p->ul); if(l > max_dist) break; for (k = a_n-1; k >= 0; k--) { lk = &(a[k]); if((lk->qe+g_gap) <= li->qs) break; if(p->v == (((lk->tn<<1)|lk->rev)^1)) { ///check if lk can be directly reachedc from li if(lk->qe > li->qs && (track[k]&((uint64_t)0x80000000))) { ; } get_rr_tse(uref, lk, &kts, &kte, &kn); ///t_dis and q_dis might be < 0 t_dis = ((int64_t)(l + kn - kte)) - ((int64_t)(in - its)); q_dis = ((int64_t)(li->qs)) - ((int64_t)(lk->qe)); } } v = p->v; } } } } int64_t gl_chain_advance(kv_ul_ov_t *res, ul_ov_t *ex, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, uint64_t *srt, uint64_t *idx, uint64_t *track, float trav_rate, void *km) { uint32_t li_v, lj_v, rev_n, gapLen = (trav_rate>0?trav_rate*qlen:0); int64_t mm_ovlp, x, i, j, k, sc, csc, mm_sc, mm_idx, qo, n_skip, n_all; ul_ov_t *li = NULL, *lj = NULL, rev_t; radix_sort_ul_ov_srt_qe(res->a, res->a + res->n); for (i = 0; i < (int64_t)res->n; ++i) { li = &(res->a[i]); li_v = (li->tn<<1)|li->rev; // if(!(li->el)) continue; mm_ovlp = uref?max_ovlp(uref->ug->g, li_v^1):max_ovlp_src(uopt, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, res->a, x); if(li->el) { csc = uref?retrieve_u_cov_region(uref, li->tn, 0, li->ts, li->te, NULL):li->te-li->ts; } else { csc = -1;///for cis overlap, the csc should be >1000; so -1 for trans overlaps should be fine } mm_sc = csc; mm_idx = -1; n_skip = n_all = 0; for (j = x; j >= 0; --j) { // collect potential destination vertices lj = &(res->a[j]); lj_v = (lj->tn<<1)|lj->rev; // if((lj->qe+gapLen) <= li->qs) break; if(lj->qe <= li->qs) break;//evan this pair has a overlap, its length will be very small; just ignore if(lj->qs >= li->qs) continue; // lj is contained in li on the query coordinate qo = infer_rovlp(li, lj, NULL, NULL); ///overlap length in query (UL read) if(li_v != lj_v && get_ecov_adv(uref, uopt, li_v^1, lj_v^1, bw, diff_ec_ul, qo)) { sc = csc + (track[j]>>32); if(sc > mm_sc) mm_sc = sc, mm_idx = j; if(res->a[j].sec == i && res->a[j].el) n_skip++; if((track[j]&((uint64_t)0x7FFFFFFF)) != ((uint64_t)0x7FFFFFFF)) { res->a[(track[j]&((uint64_t)0x7FFFFFFF))].sec = i; } track[j] |= ((uint64_t)0x80000000); } else { if(track[j]&((uint64_t)0x80000000)) track[j] -= ((uint64_t)0x80000000); } n_all++; } if(n_all > max_skip) n_all = max_skip; else n_all -= 2; //allow one mismatch; note here must be -2 if(li->el && (mm_idx<0 || n_skip0?trav_rate*qlen*li->el:0); if(gapLen > 0) { // if((lj->qe+gapLen) <= li->qs) break; ///since graph traversal just has one path, so this step might be quite easy best_path_ext(uref, uopt, gapLen, res->a, x+1, bw, diff_ec_ul, li); } } // 4294967295L track[i] = mm_sc; track[i] <<= 32; track[i] |= (mm_idx>=0?mm_idx:((uint64_t)0x7FFFFFFF)); srt[i] = mm_sc; srt[i] <<= 32; srt[i] |= i; // fprintf(stderr, "+++i:%ld, mm_idx:%ld, mm_sc:%ld\n", i, mm_idx, mm_sc); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n\n", li->tn+1, "lc"[uref->ug->u.a[li->tn].circ], li->qs, li->qe); } for (i = 0; i < (int64_t)res->n; ++i) { if(track[i]&((uint64_t)0x80000000)) track[i] -= ((uint64_t)0x80000000); } int64_t n_v, n_u, n_v0; radix_sort_gfa64(srt, srt+res->n); //ex->n = res->n; for (k = (int64_t)res->n-1, n_v = n_u = 0; k >= 0; --k) { // fprintf(stderr, "\nk:%ld\n", k); n_v0 = n_v; for (i = (uint32_t)srt[k]; i >= 0 && (track[i]&((uint64_t)0x80000000)) == 0;) { ex[n_v++] = res->a[i]; track[i] |= ((uint64_t)0x80000000); // fprintf(stderr, "+i:%ld, ", i); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n", res->a[i].tn+1, "lc"[uref->ug->u.a[res->a[i].tn].circ], res->a[i].qs, res->a[i].qe); if((track[i]&((uint64_t)0x7FFFFFFF)) == ((uint64_t)0x7FFFFFFF)) i = -1; else i = track[i]&((uint64_t)0x7FFFFFFF); } if(n_v0 == n_v) continue; ///keep the whole score; do not cut score like minigraph sc = (i<0?(srt[k]>>32):((srt[k]>>32)-(track[i]>>32))); // sc = srt[k]>>32; idx[n_u++] = ((uint64_t)sc<<32)|(n_v-n_v0); } for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { n_v0 = n_v; n_v += (uint32_t)idx[k]; res->a[k].qn = idx[k]>>32; res->a[k].ts = n_v0; res->a[k].te = n_v; rev_n = ((uint32_t)idx[k])>>1; ///we need to consider contained reads; so determining qs is not such easy res->a[k].qs = (uint32_t)-1; res->a[k].qe = ex[n_v0].qe; for (i = 0; i < rev_n; i++) { rev_t = ex[n_v0+i]; ex[n_v0+i] = ex[n_v0+rev_n-i-1]; ex[n_v0+rev_n-i-1] = rev_t; if(res->a[k].qs > ex[n_v0+i].qs) res->a[k].qs = ex[n_v0+i].qs; if(res->a[k].qs > ex[n_v0+rev_n-i-1].qs) res->a[k].qs = ex[n_v0+rev_n-i-1].qs; } if(i < ((uint32_t)idx[k]) && res->a[k].qs < ex[n_v0+i].qs) { res->a[k].qs = ex[n_v0+i].qs; } } res->n = n_u; return res->n; } **/ int64_t determine_containment_chain(const ug_opt_t *uopt, uint64_t *track, uint64_t *flag, kv_ul_ov_t *res, int32_t nc, int64_t *nsc, int64_t mm_idx, int64_t bw, double diff_ec_ul, uint32_t el, All_reads *ridx, ma_ug_t *ug) { int64_t i, k, pk, ak, e, off = 128, qo, tt = 0, ii; ul_ov_t *li = NULL, *lk = NULL; uint32_t li_v, lk_v, is_c; if(nc<=0) return 0; for (k = tt = ak = 0; k < nc; k++) { if(!(flag[res->a[k].sec]&((uint64_t)0x80000000))) { flag[res->a[k].sec] |= ((uint64_t)0x80000000); tt++; } else { res->a[ak++].sec = res->a[k].sec; } } if(tt==nc) return nsc[0] - nsc[1]; assert(ak>0); e = res->a[res->a[ak-1].sec].qe;//e is the smallest qe for (i = mm_idx, pk = 0; i >= 0;) {///i++, li->qe-- li = &(res->a[i]); li_v = (li->tn<<1)|li->rev; if((track[i]&((uint64_t)0x7FFFFFFF)) == ((uint64_t)0x7FFFFFFF)) i = -1; else i = track[i]&((uint64_t)0x7FFFFFFF); if(li->qe + off < e || tt == nc) break;//128 is used to tolerate indels; for (k = pk, ii = 0; k < ak; k++) {///k++, lk->qe-- if(res->a[k].sec == ((uint32_t)0x3FFFFFFF)) continue; lk = &(res->a[res->a[k].sec]); lk_v = (lk->tn<<1)|lk->rev; if(li->qe + off >= lk->qe) { if(ii == 0) pk = k; if(li->qs <= lk->qs + off) { qo = infer_rovlp(li, lk, NULL, NULL, ridx, ug); ///overlap length in query (UL read) if(li_v != lk_v && get_ecov_adv_back(NULL, uopt, li_v^1, lk_v^1, bw, diff_ec_ul, qo, &is_c)) { if(is_c) { tt++; res->a[k].sec = ((uint32_t)0x3FFFFFFF); if(el) nsc[0] -= ((int64_t)(lk->te-lk->ts)); else nsc[!(lk->el)] -= ((int64_t)(lk->te-lk->ts)); } } } ii = 1; } } } assert(nsc[0]>=0 && nsc[1]>=0); return nsc[0] - nsc[1]; } uint64_t push_sc_pre(int64_t mm_sc, int64_t mm_idx) { uint32_t sc = (mm_sc>=0?(((uint32_t)(mm_sc))|((uint32_t)(0x80000000))):((uint32_t)(-mm_sc))); uint64_t x = sc; x <<= 32; x |= (mm_idx>=0?mm_idx:((uint64_t)0x7FFFFFFF)); return x; } int64_t pop_sc(uint64_t x) { int64_t sc; x >>= 32; if(x&((uint64_t)(0x80000000))) { sc = x - ((uint64_t)(0x80000000)); } else { sc = x; sc *= -1; } return sc; } int64_t pop_pre(uint64_t x) { if((x&((uint64_t)0x7FFFFFFF)) == ((uint64_t)0x7FFFFFFF)) return -1; else return (x&((uint64_t)0x7FFFFFFF)); } ///mode: 0->ug; 1->read int64_t gl_chain_advance(kv_ul_ov_t *res, ul_ov_t *ex, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, uint64_t *srt, uint64_t *idx, uint64_t *track, int64_t trans_sc, uint64_t mode, All_reads *ridx, ma_ug_t *ug, int64_t debug_i, void *km) { // fprintf(stderr, "\n+++[M::%s] res->n:%u\n", __func__, (uint32_t)res->n); if(res->n == 0) return 0; uint32_t li_v, lj_v, rev_n; int64_t mm_ovlp, x, i, j, k, sc, csc, mm_sc, mm_idx, qo, share, n_el = 0; ul_ov_t *li = NULL, *lj = NULL, rev_t; radix_sort_ul_ov_srt_qe(res->a, res->a + res->n); for (i = 1, j = 0; i <= (int64_t)res->n; i++) { if (i == (int64_t)res->n || res->a[i].qe != res->a[j].qe) { if(i - j > 1) { radix_sort_ul_ov_srt_qs(res->a+j, res->a+i); } j = i; } } for (i = 0; i < (int64_t)res->n; ++i) { li = &(res->a[i]); li_v = (li->tn<<1)|li->rev; mm_ovlp = mode?max_ovlp_src(uopt, li_v^1):max_ovlp(uref->ug->g, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, res->a, x+G_CHAIN_INDEL); if(li->el) csc = mode?retrieve_r_cov_region(uref, li->tn, 0, li->ts, li->te, NULL):retrieve_u_cov_region(uref, li->tn, 0, li->ts, li->te, NULL); else csc = trans_sc; //trans overlaps mm_sc = csc; mm_idx = -1; // if(i == 37 || i == 36 || i == 35 || i == 32) fprintf(stderr, "*i:%ld, x:%ld, mm_sc:%ld\n", i, x, mm_sc); for (j = x; j >= 0; --j) { // collect potential destination vertices lj = &(res->a[j]); lj_v = (lj->tn<<1)|lj->rev; // if((lj->qe+gapLen) <= li->qs) break; if(lj->qe+G_CHAIN_INDEL <= li->qs) break;//even this pair has a overlap, its length will be very small; just ignore if(lj->qs >= li->qs+G_CHAIN_INDEL) continue; // lj is contained in li on the query coordinate; 128 for indel offset qo = infer_rovlp(li, lj, NULL, NULL, ridx, ug); ///overlap length in query (UL read) // if(i == 37 || i == 36 || i == 35 || i == 32) fprintf(stderr, ">i:%ld, j:%ld, qo:%ld\n", i, j, qo); if(li_v != lj_v && get_ecov_adv(uref, uopt, li_v^1, lj_v^1, bw, diff_ec_ul, qo, mode, &share)) { // if(i == 37 || i == 36 || i == 35 || i == 32) fprintf(stderr, "#i:%ld, j:%ld, share:%ld\n", i, j, share); sc = csc + pop_sc(track[j]); // if((!mode)&&i==11&&j==10) { // fprintf(stderr,"+share:%ld, i:%ld, j:%ld, li_v^1:%u, lj_v^1:%u\n", // share, i, j, li_v^1, lj_v^1); // } // if((mode&&i==21&&j==20) || (mode&&i==22&&j==21) || (mode&&i==23&&j==22)) { // fprintf(stderr,"-share:%ld, i:%ld, j:%ld, li_v^1:%u, lj_v^1:%u\n", // share, i, j, li_v^1, lj_v^1); // } if(li->el && lj->el) sc -= (share>=csc?csc:share);///csc must be larger than 0 // if((!li->el) && (!lj->el)) sc -= ((share>=o_csc?o_csc:share)*(-trans_scl)); if(sc > mm_sc) mm_sc = sc, mm_idx = j; } } track[i] = push_sc_pre(mm_sc, mm_idx); srt[i] = track[i]>>32; srt[i] <<= 32; srt[i] |= i; n_el += li->el; // if(mode) { // fprintf(stderr, "[M::%.*s] i:%ld, li->el:%u, li->score:%ld (raw_sc:%u), mm_idx:%ld, mm_sc:%ld, q[%u, %u), t[%u, %u), rev:%c\n", // (int32_t)Get_NAME_LENGTH(R_INF, li->tn), Get_NAME(R_INF, li->tn), i, li->el, csc, li->te - li->ts, mm_idx, mm_sc, li->qs, li->qe, li->ts, li->te, "+-"[li->rev]); // } else { // fprintf(stderr, "[M::utg%.6u%c] i:%ld, li->el:%u, li->score:%ld (raw_sc:%u), mm_idx:%ld, mm_sc:%ld, q[%u, %u), t[%u, %u), rev:%c\n", // li->tn+1, "lc"[uref->ug->u.a[li->tn].circ], i, li->el, csc, li->te - li->ts, mm_idx, mm_sc, li->qs, li->qe, li->ts, li->te, "+-"[li->rev]); // } // if(!mode) { // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n", li->tn+1, "lc"[uref->ug->u.a[li->tn].circ], li->qs, li->qe); // } } int64_t n_v, n_u, n_v0, le, lnv; radix_sort_gfa64(srt, srt+res->n); for (k = (int64_t)res->n-1, n_v = n_u = 0; k >= 0; --k) { n_v0 = n_v; i = (uint32_t)srt[k]; if(res->a[i].el) { ///chain must start from cis alignments for (le = -1; i >= 0 && (track[i]&((uint64_t)0x80000000)) == 0;) { if(res->a[i].el) { le = -1; }else if(n_v>n_v0 && ex[n_v-1].el) { le = i; lnv = n_v;///cut the cis alignments in the end } ex[n_v++] = res->a[i]; track[i] |= ((uint64_t)0x80000000); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n", res->a[i].tn+1, "lc"[uref->ug->u.a[res->a[i].tn].circ], res->a[i].qs, res->a[i].qe); i = pop_pre(track[i]); } } if(n_v0 == n_v) continue; if(le >= 0) { i = le; n_v = lnv; } if(n_v0 == n_v) continue; // fprintf(stderr, "[++chain::] beg_idx->%u, end_idx->%ld, le->%ld, chain_n->%ld\n", (uint32_t)srt[k], i, le, n_v - n_v0); ///keep the whole score; do not cut score like minigraph // sc = pop_sc(srt[k]); sc = (i<0?(pop_sc(srt[k])):(pop_sc(srt[k])-pop_sc(track[i]))); // fprintf(stderr, "++[M::%s] k:%ld, n_v0:%ld, n_v:%ld, le:%ld, sc:%ld, beg:%u, end:%ld, p_score:%ld, cut_score:%ld\n", // __func__, k, n_v0, n_v, le, sc, (uint32_t)srt[k], i, pop_sc(srt[k]), i<0?0:pop_sc(track[i])); if(sc /**<=**/< 0) {///sc might be 0, if the UL alignment cannot cover the whole overlap between two HiFi reads n_v = n_v0; continue; } // idx[n_u++] = push_sc_pre(sc, n_v-n_v0); idx[n_u++] = ((uint64_t)sc<<32)|(n_v-n_v0); } // fprintf(stderr, "[M::%s] n_u:%ld, n_v:%ld\n", __func__, n_u, n_v); for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { n_v0 = n_v; n_v += (uint32_t)idx[k]; // fprintf(stderr, "[M::%s] k:%ld, n_v0:%ld, n_v:%ld\n", __func__, k, n_v0, n_v); res->a[k].qn = idx[k]>>32;//score res->a[k].ts = n_v0; res->a[k].te = n_v;///idx rev_n = ((uint32_t)idx[k])>>1; ///we need to consider contained reads; so determining qs is not such easy res->a[k].qs = (uint32_t)-1; res->a[k].qe = ex[n_v0].qe; for (i = 0; i < rev_n; i++) { rev_t = ex[n_v0+i]; ex[n_v0+i] = ex[n_v-i-1]; ex[n_v-i-1] = rev_t; if(res->a[k].qs > ex[n_v0+i].qs) res->a[k].qs = ex[n_v0+i].qs; if(res->a[k].qs > ex[n_v-i-1].qs) res->a[k].qs = ex[n_v-i-1].qs; n_el -= ex[n_v0+i].el; n_el -= ex[n_v-i-1].el; } if(((uint32_t)idx[k])&1) { if(res->a[k].qs > ex[n_v0+i].qs) res->a[k].qs = ex[n_v0+i].qs; n_el -= ex[n_v0+i].el; } assert(ex[n_v0].el && ex[n_v-1].el); // fprintf(stderr, "[M::%s] k:%ld, qs:%u, qe:%u, chain_occ:%u, chain_score:%u\n", __func__, k, // res->a[k].qs, res->a[k].qe, res->a[k].te - res->a[k].ts, res->a[k].qn); } // if(n_el) { // fprintf(stderr, "[M::%s] debug_i->%ld, n_el->%ld, n_u->%ld, n_v->%ld\n", __func__, debug_i, n_el, n_u, n_v); // } assert(n_el == 0); res->n = n_u; radix_sort_ul_ov_srt_qn(res->a, res->a + res->n);//sort by score // fprintf(stderr, "---[M::%s] n_u:%ld, n_v:%ld\n", __func__, n_u, n_v); return n_v; } int64_t gl_chain_advance_back(kv_ul_ov_t *res, ul_ov_t *ex, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, uint64_t *srt, uint64_t *idx, uint64_t *track, float trans_allow, All_reads *ridx, ma_ug_t *ug, void *km) { uint32_t li_v, lj_v, rev_n, is_c, nc, s_nc; int64_t mm_ovlp, x, i, j, k, sc, csc, mm_sc, mm_idx, qo, trans_scl = (int64_t)(((float)(1))/trans_allow), nsc[2]; ul_ov_t *li = NULL, *lj = NULL, rev_t; radix_sort_ul_ov_srt_qe(res->a, res->a + res->n); for (i = s_nc = 0; i < (int64_t)res->n; ++i) { li = &(res->a[i]); li_v = (li->tn<<1)|li->rev; mm_ovlp = uref?max_ovlp(uref->ug->g, li_v^1):max_ovlp_src(uopt, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, res->a, x); csc = uref?retrieve_u_cov_region(uref, li->tn, 0, li->ts, li->te, NULL):li->te-li->ts; if(!(li->el)) { csc *= -trans_scl; //trans overlaps if(csc >= 0) csc = -1; } mm_sc = csc; mm_idx = -1; nc = nsc[0] = nsc[1] = 0; for (j = x; j >= 0; --j) { // collect potential destination vertices lj = &(res->a[j]); lj_v = (lj->tn<<1)|lj->rev; // if((lj->qe+gapLen) <= li->qs) break; if(lj->qe <= li->qs) break;//even this pair has a overlap, its length will be very small; just ignore // if(lj->qs >= li->qs) continue; // lj is contained in li on the query coordinate qo = infer_rovlp(li, lj, NULL, NULL, ridx, ug); ///overlap length in query (UL read) if(li_v != lj_v && get_ecov_adv_back(uref, uopt, li_v^1, lj_v^1, bw, diff_ec_ul, qo, &is_c)) { if(!is_c) { sc = csc + pop_sc(track[j]); if(sc > mm_sc) mm_sc = sc, mm_idx = j; } else if(!uref) {///with uref, retrieve_u_cov_region has already consider contained reads res->a[nc++].sec = j; if(li->el) nsc[0] += lj->te-lj->ts; else nsc[!(lj->el)] += lj->te-lj->ts; } } } if(nc && (!uref) && mm_idx>=0) {///deal with containments mm_sc += determine_containment_chain(uopt, track, srt, res, nc, nsc, mm_idx, bw, diff_ec_ul, li->el, ridx, ug); s_nc++; } track[i] = push_sc_pre(mm_sc, mm_idx); srt[i] = track[i]>>32; srt[i] <<= 32; srt[i] |= i; // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n\n", li->tn+1, "lc"[uref->ug->u.a[li->tn].circ], li->qs, li->qe); } if(s_nc) { for (i = 0; i < (int64_t)res->n; ++i) { if(srt[i]&((uint64_t)0x80000000)) srt[i]-=((uint64_t)0x80000000); } } int64_t n_v, n_u, n_v0, le, lnv; radix_sort_gfa64(srt, srt+res->n); //ex->n = res->n; for (k = (int64_t)res->n-1, n_v = n_u = 0; k >= 0; --k) { n_v0 = n_v; i = (uint32_t)srt[k]; if(i>=0 && (res->a[i].el)) { ///chain must start from cis alignments for (le = -1; i >= 0 && (track[i]&((uint64_t)0x80000000)) == 0;) { if(res->a[i].el) { le = -1; }else if(n_v>n_v0 && ex[n_v-1].el) { le = i; lnv = n_v;///cut the cis alignments in the end } ex[n_v++] = res->a[i]; track[i] |= ((uint64_t)0x80000000); // fprintf(stderr, "[M::utg%.6d%c] qs->%u; qe->%u\n", res->a[i].tn+1, "lc"[uref->ug->u.a[res->a[i].tn].circ], res->a[i].qs, res->a[i].qe); i = pop_pre(track[i]); } } if(n_v0 == n_v) continue; if(le >= 0) { i = le; n_v = lnv; } if(n_v0 == n_v) continue; ///keep the whole score; do not cut score like minigraph // sc = pop_sc(srt[k]); sc = (i<0?(pop_sc(srt[k])):(pop_sc(srt[k])-pop_sc(track[i]))); if(sc < 0) { n_v = n_v0; continue; } // idx[n_u++] = push_sc_pre(sc, n_v-n_v0); idx[n_u++] = ((uint64_t)sc<<32)|(n_v-n_v0); } for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { n_v0 = n_v; n_v += (uint32_t)idx[k]; res->a[k].qn = idx[k]>>32; res->a[k].ts = n_v0; res->a[k].te = n_v; rev_n = ((uint32_t)idx[k])>>1; ///we need to consider contained reads; so determining qs is not such easy res->a[k].qs = (uint32_t)-1; res->a[k].qe = ex[n_v0].qe; for (i = 0; i < rev_n; i++) { rev_t = ex[n_v0+i]; ex[n_v0+i] = ex[n_v0+rev_n-i-1]; ex[n_v0+rev_n-i-1] = rev_t; if(res->a[k].qs > ex[n_v0+i].qs) res->a[k].qs = ex[n_v0+i].qs; if(res->a[k].qs > ex[n_v0+rev_n-i-1].qs) res->a[k].qs = ex[n_v0+rev_n-i-1].qs; } if(i < ((uint32_t)idx[k]) && res->a[k].qs < ex[n_v0+i].qs) { res->a[k].qs = ex[n_v0+i].qs; } } res->n = n_u; radix_sort_ul_ov_srt_qn(res->a, res->a + res->n); return n_v; } uint32_t check_trans_rate(ul_ov_t *a, int64_t a_n, float trans_thres) { uint32_t sp = (uint32_t)-1, ep = (uint32_t)-1, tts = (uint32_t)-1, tte = 0, el = 0, iel = 0; int64_t k; for (k = a_n-1; k >= 0; k--) { if(a[k].qs < tts) tts = a[k].qs; if(a[k].qe > tte) tte = a[k].qe; if(!(a[k].el)) continue; if(sp == (uint32_t)-1 || a[k].qe <= sp) { if(sp != (uint32_t)-1) el += ep - sp; sp = a[k].qs; ep = a[k].qe; } else { sp = MIN(sp, a[k].qs); } } if(sp != (uint32_t)-1) el += ep - sp; iel = (tte - tts) - el; // fprintf(stderr, "[M::%s] el:%u, iel:%u\n", __func__, el, iel); if((iel == 0) || (iel <= ((tte - tts)*trans_thres))) return 1; return 0; } uint32_t ff_chain(kv_ul_ov_t *idx, int64_t qlen, float cov_rate, float trans_thres, ul_ov_t *a, overlap_region_alloc* olist, haplotype_evdience_alloc *hap, const ul_idx_t *uref, double diff_ec_ul, int64_t winLen, void *km) { if(idx->n <= 0) return 0; ul_ov_t *m = &(idx->a[idx->n-1]); //largest chain // fprintf(stderr, "[M::%s] m->score:%u, m->qs:%u, m->qe:%u, chain_n:%u\n", __func__, m->qn, m->qs, m->qe, m->te-m->ts); if((m->qe-m->qs) <= (qlen*cov_rate)) return 0; if(check_trans_rate(a+m->ts, m->te-m->ts, trans_thres)) return 1; if(olist && hap && uref) { int64_t idx_n = idx->n, z, i, het_n, resc_tk = 0, f = 0; uint64_t si; ma_utg_t *u = NULL; for (z = m->ts; z < m->te; z++) { if(a[z].el) { kv_push_km(km, ul_ov_t, *idx, a[z]); } else { i = a[z].qn; si = 0; het_n = update_ava_het_site(hap, i, &si, NULL, 1); assert(het_n > 0 && olist->list[i].is_match == 2); u = &(uref->ug->u.a[olist->list[i].y_id]); if(u->n > 1) { resc_tk += rescue_trans_ul_chains(uref, &(olist->list[i]), hap->list+si, het_n, u, idx, diff_ec_ul, winLen, 0, NULL, km); } } } if(resc_tk) { radix_sort_ul_ov_srt_qe(idx->a+idx_n, idx->a+idx->n); f = check_trans_rate(idx->a+idx_n, idx->n-idx_n, trans_thres); } idx->n = idx_n; return f; } return 0; } void dump_chain(kv_ul_ov_t *des, ul_ov_t *src, ul_ov_t *chain, void *km) { ///note: dump results to may change , so we should save in advance uint64_t beg = chain->ts, occ = chain->te - chain->ts; kv_resize_km(km, ul_ov_t, *des, occ); des->n = occ; memcpy(des->a, src + beg, occ*sizeof((*src))); } int64_t dedup_sort_contains(ul_ov_t *a, int64_t a_n, ul_contain *ct, const ug_opt_t *uopt) { int64_t k, l, ci; ul_ov_t *z = NULL; for (k = 0; k < a_n; k++) { z = &(a[k]); if(z->tn&((uint32_t)(0x80000000))) continue;///contained alignment if(ct->is_c.a[z->tn] == 0) continue; for (ci = k+1; ci < a_n; ci++) { if(a[ci].qe > z->qe + G_CHAIN_INDEL) break;///128 is for indel if(a[ci].qn == (uint32_t)-1) continue; if(!(a[ci].tn&((uint32_t)(0x80000000)))) continue; if(z->qs <= a[ci].qs + G_CHAIN_INDEL && z->qe + G_CHAIN_INDEL >= a[ci].qe) { if(check_contain_pair(uopt, (a[ci].tn<<1)>>1, z->tn, 1)) { a[ci].qn = (uint32_t)-1; } } } for (ci = k-1; ci >= 0; ci--) { if(a[ci].qe + G_CHAIN_INDEL <= z->qs) break; if(a[ci].qn == (uint32_t)-1) continue; if(!(a[ci].tn&((uint32_t)(0x80000000)))) continue; if(z->qs <= a[ci].qs + G_CHAIN_INDEL && z->qe + G_CHAIN_INDEL >= a[ci].qe) { if(check_contain_pair(uopt, (a[ci].tn<<1)>>1, z->tn, 1)) { a[ci].qn = (uint32_t)-1; } } } } for (k = l = 0; k < a_n; k++) { if(a[k].qn == (uint32_t)-1) continue; if(k != l) a[l] = a[k]; a[l].tn <<= 1; a[l].tn >>= 1; ++l; } return l; } void ins_merge_ul_ov(kv_ul_ov_t *idx, int64_t idx_s, int64_t idx_e, ul_ov_t q) { int64_t k, ii, s = -1, e = -1, ovlp = 0, qs = q.qs, qe = q.qe; for (k = idx_s, ii = -1; k < idx_e; k++) { if(ii == -1 && q.qs > idx->a[k].qs) ii = k; if(((int64_t)(idx->a[k].qs)) >= e) { if(s >= 0 && e >= 0) { ovlp += ((MIN(e, qe) > MAX(s, qs))?(MIN(e, qe) - MAX(s, qs)):0); } s = idx->a[k].qs; e = idx->a[k].qe; } else { if(e < ((int64_t)(idx->a[k].qe))) e = idx->a[k].qe; } } if(s >= 0 && e >= 0) { ovlp += ((MIN(e, qe) > MAX(s, qs))?(MIN(e, qe) - MAX(s, qs)):0); } } void dump_all_chain(kv_ul_ov_t *idx, kv_ul_ov_t *ax, int64_t ax_new_occ, int64_t qlen, float primary_cov_rate, float primary_score_rate) { if(idx->n <= 0) return; ul_ov_t *m = &(idx->a[idx->n-1]); //largest chain ul_ov_t *a = ax->a + ax->n; int64_t k, i, z, l, idx_n = idx->n; uint64_t ovlp; if((m->qe-m->qs) > (qlen*primary_cov_rate)) { ///found a primary chain for (k = m->ts, l = 0; k < m->te; k++) { a[l] = a[k]; a[l].tn |= ((uint32_t)(0x80000000)); l++; } ax->n += l; } else { for (k = idx_n-1; k >= 0; k--) { for (i = idx_n-1; i > k; i--) { if(idx->a[i].qn == (uint32_t)-1) continue;///just remove totally contained alignments if(idx->a[k].qn > idx->a[i].qn*primary_score_rate) continue;///consider score ovlp = ((MIN(idx->a[k].qe, idx->a[i].qe) > MAX(idx->a[k].qs, idx->a[i].qs))? (MIN(idx->a[k].qe, idx->a[i].qe) - MAX(idx->a[k].qs, idx->a[i].qs)):0); if(ovlp > ((idx->a[k].qe-idx->a[k].qs)*primary_cov_rate)) { for (z = idx->a[k].ts; z < idx->a[k].te; z++) a[z].el = 1; idx->a[k].qn = (uint32_t)-1; break; } } // ins_merge_ul_ov(idx, idx_n, idx->n, idx->a[k]); } for (k = 0, l = 0; k < ax_new_occ; k++) { if(a[k].el) continue; a[l] = a[k]; l++; } radix_sort_ul_ov_srt_qe(a, a + l); ax->n += l; } } void dump_all_chain_simple(kv_ul_ov_t *idx, kv_ul_ov_t *ax, int64_t ax_new_occ, int64_t qlen, float primary_cov_rate, float fragement_cov_rate, float primary_fragment_cov_rate, float primary_fragment_second_score_rate, float trans_thres) { if(idx->n <= 0) return; ul_ov_t *m = &(idx->a[idx->n-1]); //largest chain ul_ov_t *a = ax->a + ax->n; int64_t k, z, l, idx_n = idx->n, ovlp, om, ok; // fprintf(stderr, "[M::%s] m->score:%u, m->qs:%u, m->qe:%u, chain_n:%u\n", __func__, m->qn, m->qs, m->qe, m->te-m->ts); if(((m->qe-m->qs) > (qlen*primary_cov_rate)) && (check_trans_rate(a+m->ts, m->te-m->ts, trans_thres))) { ///found a primary chain for (k = m->ts, l = 0; k < m->te; k++) { a[l] = a[k]; a[l].tn |= ((uint32_t)(0x80000000)); a[l].el = 1; l++; } ax->n += l; } else { if(((m->qe-m->qs) > (qlen*primary_fragment_cov_rate)) && (check_trans_rate(a+m->ts, m->te-m->ts, trans_thres))) { om = m->qe - m->qs; for (k = 0; k < idx_n-1; k++) { ovlp = ((MIN((m->qe), (idx->a[k].qe)) > MAX((m->qs), (idx->a[k].qs)))? (MIN((m->qe), (idx->a[k].qe)) - MAX((m->qs), (idx->a[k].qs))):0); if(ovlp == 0) continue; ok = idx->a[k].qe - idx->a[k].qs; if(ok > om ) ok = om; if((ovlp > ok*0.25) && idx->a[k].qn > (m->qn*primary_fragment_second_score_rate)) break; } if(k >= idx_n-1) { for (k = m->ts; k < m->te; k++) { if(a[k].el) a[k].tn |= ((uint32_t)(0x80000000)); } } } radix_sort_ul_ov_srt_qe(idx->a, idx->a + idx->n); for (k = 0; k < idx_n; k++) { if(k < idx_n-1 && idx->a[k].qe > idx->a[k+1].qs) break;//not one chain if((idx->a[k].qe - idx->a[k].qs) > (qlen*fragement_cov_rate)) {///large enough fragements if(!check_trans_rate(a+idx->a[k].ts, idx->a[k].te-idx->a[k].ts, trans_thres)) break; } } if(k == idx_n) {///only if there is a clear chain (with holes) for (k = 0; k < idx_n; k++) { if((idx->a[k].qe - idx->a[k].qs) <= (qlen*fragement_cov_rate)) continue; for (z = idx->a[k].ts; z < idx->a[k].te; z++) { if(a[z].el) a[z].tn |= ((uint32_t)(0x80000000)); } } } /** for (k = 0, l = 0; k < ax_new_occ; k++) { if(!(a[k].el)) continue; a[l] = a[k]; l++; } radix_sort_ul_ov_srt_qe(a, a + l); ax->n += l; **/ radix_sort_ul_ov_srt_qe(a, a + ax_new_occ); for (k = 1, l = 0; k <= ax_new_occ; k++) { if (k == ax_new_occ || a[k].qe != a[l].qe) { if(k - l > 1) radix_sort_ul_ov_srt_qs(a+l, a+k); l = k; } } ax->n += ax_new_occ; } } void save_tmp_chains(ul_ov_t *idx_a, uint64_t idx_n, uint64_t *idx_buf_0, uint64_t *idx_buf_1, ul_ov_t *cc_a, uint64_t cc_n, uint64_t *cc_buf) { uint64_t k; for (k = 0; k < idx_n; k++) ; } void debug_reverse_chain(ul_ov_t *a, int64_t a_n) { int64_t rev_n = a_n>>1, i; ul_ov_t rev_t; for (i = 0; i < rev_n; i++) { rev_t = a[i]; a[i] = a[a_n-i-1]; a[a_n-i-1] = rev_t; } } int64_t gl_chain_refine_advance(overlap_region_alloc* olist, Correct_dumy* dumy, haplotype_evdience_alloc *hap, glchain_t *ll, const ul_idx_t *uref, double diff_ec_ul, int64_t winLen, int64_t qlen, const ug_opt_t *uopt, int64_t debug_i, void *km) { // ll->tk.n = ll->lo.n = 0; kv_ul_ov_t *idx = &(ll->lo); ul_contain *ct = uref->ct; uint64_t o2 = gl_chain_gen(olist, uref, idx, 0, km); if(idx->n == 0) return 0; // fprintf(stderr, "[M::%s] qlen:%ld, idx->n:%u\n", __func__, qlen, (uint32_t)idx->n); uint64_t k, an, cn, si = 0, ei = 0, resc = 0, resc_tk = 0, tk_pl = 0, f = 0, occ = 0, cis_occ = 0, t_cis = 0; ma_utg_t *u = NULL; overlap_region *o = NULL; kv_resize_km(km, uint64_t, ll->srt.a, idx->n); kv_resize_km(km, uint64_t, hap->snp_srt, idx->n); kv_resize_km(km, ul_ov_t, ll->tk, ll->tk.n+idx->n); ///chain exact U-matches occ = gl_chain_advance(idx, ll->tk.a+ll->tk.n, uref, uopt, G_CHAIN_BW, diff_ec_ul, qlen, UG_SKIP, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, G_CHAIN_TRANS_WEIGHT, 0, NULL, uref->ug, debug_i, km); if(occ) { if(ff_chain(idx, qlen, P_CHAIN_COV, G_CHAIN_TRANS_RATE, ll->tk.a+ll->tk.n, NULL, NULL, NULL, diff_ec_ul, winLen, km)) { f = 1; //dump_chain(idx, ll->tk.a+ll->tk.n, &(idx->a[idx->n-1]), km); for (k = idx->a[idx->n-1].ts; k < idx->a[idx->n-1].te; k++) { olist->list[ll->tk.a[ll->tk.n+k].qn].x_pos_strand = 1; } } else if(o2) {///means there are trans overlaps gl_chain_gen(olist, uref, idx, 1, km); kv_resize_km(km, uint64_t, ll->srt.a, idx->n); kv_resize_km(km, uint64_t, hap->snp_srt, idx->n); kv_resize_km(km, ul_ov_t, ll->tk, ll->tk.n+idx->n); ///chain all U-matches occ = gl_chain_advance(idx, ll->tk.a+ll->tk.n, uref, uopt, G_CHAIN_BW, diff_ec_ul, qlen, UG_SKIP, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, G_CHAIN_TRANS_WEIGHT, 0, NULL, uref->ug, debug_i, km); if(ff_chain(idx, qlen, P_CHAIN_COV, G_CHAIN_TRANS_RATE, ll->tk.a+ll->tk.n, olist, hap, uref, diff_ec_ul, winLen, km)) { f = 1; //dump_chain(idx, ll->tk.a+ll->tk.n, &(idx->a[idx->n-1]), km); for (k = idx->a[idx->n-1].ts; k < idx->a[idx->n-1].te; k++) { olist->list[ll->tk.a[ll->tk.n+k].qn].x_pos_strand = 1; } } } } if(!f) {///if f == 1, only dump primary chain; otherwise dump all chains ///we can save all data to buffer like ll->srt.a in advance; in case we don't need third round of chaining ///means no trans overlaps, no need to do third round of chaining // if(!o2) { // ; // } /** uint64_t z; for (k = 0; k < idx->n; k++) { if(check_trans_rate(ll->tk.a+ll->tk.n+idx->a[k].ts, idx->a[k].te-idx->a[k].ts, G_CHAIN_TRANS_RATE)) { for (z = idx->a[k].ts; z < idx->a[k].te; z++) { olist->list[ll->tk.a[ll->tk.n+z].qn].x_pos_strand = 1; } } else { for (z = idx->a[k].ts; z < idx->a[k].te; z++) { if(!(ll->tk.a[ll->tk.n+z].el)) continue; olist->list[ll->tk.a[ll->tk.n+z].qn].x_pos_strand = 1; } } } **/ for (k = 0; k < occ; k++) { olist->list[ll->tk.a[ll->tk.n+k].qn].x_pos_strand = 1; } // kv_resize_km(km, ul_ov_t, *idx, occ); idx->n = occ; // memcpy(idx->a, ll->tk.a+ll->tk.n, occ*sizeof((*(idx->a)))); } // for (k = 0; k < idx->n; k++) olist->list[idx->a[k].qn].x_pos_strand = 1; for (k = 0, idx->n = 0, tk_pl = ll->tk.n, t_cis = 0; k < olist->length; k++) { o = &(olist->list[k]); ///if f == 1, no matter if(o->x_pos_strand && (f || o->is_match == 1)){ u = &(uref->ug->u.a[o->y_id]);///overlaped reads resc_tk += rescue_trans_ul_chains(uref, o, NULL, 0, u, &(ll->tk), -1, -1, 0, NULL, km); } else if((!f) && o->is_match == 2) { // fprintf(stderr, "###[M::%s] # k:%lu, # o->y_id:%u\n", __func__, k, o->y_id); an = update_ava_het_site(hap, k, &si, &ei, 1); // if(an != get_het_site(hap, k)) fprintf(stderr, "an->%lu, get_het_site->%lu\n", an, get_het_site(hap, k)); assert(an > 0); cn = ((uint32_t)(ct->idx.a[o->y_id])); if(cn > 0) { resc += rescue_contain_ul_chains(uref, o, hap->list+si, an, ct->rids.a + ((ct->idx.a[o->y_id])>>32), cn, idx, diff_ec_ul, winLen, 0, km); } u = &(uref->ug->u.a[o->y_id]); if(u->n > 1 || o->x_pos_strand) {///no redundant items here resc_tk += rescue_trans_ul_chains(uref, o, hap->list+si, an, u, &(ll->tk), diff_ec_ul, winLen, o->x_pos_strand, &cis_occ, km); t_cis += cis_occ; } si = ei; } } assert(ll->tk.n == resc_tk+tk_pl); assert(idx->n == resc); if(f) assert(resc==0); if(!f) {///dedup contained alignments if(idx->n) {///if some contained alignments have been rescued radix_sort_ul_ov_srt_tn(idx->a, idx->a + idx->n); idx->n = dedup_sort_ul_ov_t(idx->a, idx->n);///different trans alignments may have the same contained alignment } resc = idx->n; // fprintf(stderr, "***[M::%s] # contain:%lu, # non-contain:%lu\n", __func__, resc, (uint64_t)(ll->tk.n-tk_pl)); for (k = tk_pl; k < ll->tk.n; k++) {///dump all non-contained reads kv_push_km(km, ul_ov_t, *idx, ll->tk.a[k]); if(idx->a[idx->n-1].tn&((uint32_t)(0x80000000))) { idx->a[idx->n-1].tn -= ((uint32_t)(0x80000000)); } } ll->tk.n = tk_pl; radix_sort_ul_ov_srt_qe(idx->a, idx->a + idx->n); if(resc) {///need to dedup contained alignment again // fprintf(stderr, "[M::%s] idx->n:%lu, resc:%lu\n", __func__, (uint64_t)idx->n, resc); idx->n = dedup_sort_contains(idx->a, idx->n, ct, uopt); } kv_resize_km(km, uint64_t, ll->srt.a, idx->n); kv_resize_km(km, uint64_t, hap->snp_srt, idx->n); kv_resize_km(km, ul_ov_t, ll->tk, ll->tk.n+idx->n); occ = gl_chain_advance(idx, ll->tk.a+ll->tk.n, uref, uopt, G_CHAIN_BW, diff_ec_ul, qlen, UG_SKIP, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, G_CHAIN_TRANS_WEIGHT, 1, &R_INF, NULL, debug_i, km); // fprintf(stderr, "***[M::%s] ll->tk.n:%u, occ:%lu\n", __func__, (uint32_t)ll->tk.n, occ); dump_all_chain_simple(idx, &(ll->tk), occ, qlen, P_CHAIN_COV, P_FRAGEMENT_CHAIN_COV, P_FRAGEMENT_PRIMARY_CHAIN_COV, P_FRAGEMENT_PRIMARY_SECOND_COV, G_CHAIN_TRANS_RATE); // fprintf(stderr, ">>>[M::%s] ll->tk.n:%u\n", __func__, (uint32_t)ll->tk.n); // dump_all_chain(idx, &(ll->tk), occ, qlen, P_CHAIN_COV, P_CHAIN_SCORE); } else { ///for primary chain, each element x: (x->tn & (uint32_t)(0x80000000)) radix_sort_ul_ov_srt_qe(ll->tk.a+tk_pl, ll->tk.a+ll->tk.n); } // debug_reverse_chain(ll->tk.a+tk_pl, ll->tk.n-tk_pl); /** if(idx->n > 0) { an = infer_read_ovlp(uref, olist, idx , &(ll->tk), diff_ec_ul, winLen, uopt, ct, km); // if(an) fill_edge_weight(ll->tk.a+ll->tk.n-an, an, uopt, G_CHAIN_BW, diff_ec_ul, qlen); } **/ return 1; } uint64_t kv_ul_ov_t_statistics(kv_ul_ov_t *olist, uint64_t qn, int64_t *occ) { int64_t k, l = 0; uint32_t sp = (uint32_t)-1, ep = (uint32_t)-1; for (k = olist->n-1; k >= 0 && olist->a[k].qn == qn; k--) { if(!(olist->a[k].el)) continue; if(sp == (uint32_t)-1 || olist->a[k].qe <= sp) { if(sp != (uint32_t)-1) l += ep - sp; sp = olist->a[k].qs; ep = olist->a[k].qe; } else { sp = MIN(sp, olist->a[k].qs); } (*occ)++; } if(sp != (uint32_t)-1) l += ep - sp; return l; } static void worker_for_ul_scall_alignment(void *data, long i, int tid) // callback for kt_for() { utepdat_t *s = (utepdat_t*)data; ha_ovec_buf_t *b = s->hab[tid]; glchain_t *bl = &(s->ll[tid]); int64_t /**rid = s->id+i,**/ winLen = MIN((((double)THRESHOLD_MAX_SIZE)/s->opt->diff_ec_ul), WINDOW); uint64_t align = 0; int fully_cov, abnormal; void *km = s->buf?(s->buf[tid]?s->buf[tid]->km:NULL):NULL; // if(s->id+i!=23) return; // fprintf(stderr, "[M::%s] rid:%ld\n", __func__, s->id+i); // if (memcmp(UL_INF.nid.a[s->id+i].a, "d0aab024-b3a7-40fb-83cc-22c3d6d951f8", UL_INF.nid.a[s->id+i].n-1)) return; // fprintf(stderr, "[M::%s::] ==> len: %lu\n", __func__, s->len[i]); ha_get_ul_candidates_interface(b->abl, i, s->seq[i], s->len[i], s->opt->w, s->opt->k, s->uu, &b->olist, &b->olist_hp, &b->clist, s->opt->bw_thres, s->opt->max_n_chain, 1, &(b->k_flag), &b->r_buf, &(b->tmp_region), NULL, &(b->sp), km); clear_Cigar_record(&b->cigar1); clear_Round2_alignment(&b->round2); // return; // b->num_correct_base += overlap_statistics(&b->olist, NULL, 0); b->self_read.seq = s->seq[i]; b->self_read.length = s->len[i]; b->self_read.size = 0; correct_ul_overlap(&b->olist, s->uu, &b->self_read, &b->correct, &b->ovlp_read, &b->POA_Graph, &b->DAGCon, &b->cigar1, &b->hap, &b->round2, 0, 1, &fully_cov, &abnormal, s->opt->diff_ec_ul, winLen, km); // uint64_t k; // for (k = 0; k < b->olist.length; k++) { // if(b->olist.list[k].is_match == 1) b->num_correct_base += b->olist.list[k].x_pos_e+1-b->olist.list[k].x_pos_s; // if(b->olist.list[k].is_match == 2) b->num_recorrect_base += b->olist.list[k].x_pos_e+1-b->olist.list[k].x_pos_s; // } // gl_chain_refine(&b->olist, &b->correct, &b->hap, bl, s->uu, s->opt->diff_ec_ul, winLen, s->len[i], km); gl_chain_refine_advance(&b->olist, &b->correct, &b->hap, bl, s->uu, s->opt->diff_ec_ul, winLen, s->len[i], s->uopt, s->id+i, km); // return; // b->num_read_base += b->self_read.length; // b->num_correct_base += b->correct.corrected_base; // b->num_recorrect_base += b->round2.dumy.corrected_base; memset(&b->self_read, 0, sizeof(b->self_read)); align = kv_ul_ov_t_statistics(&(bl->tk), i, &(b->num_recorrect_base)); if(align == s->len[i]) { free(s->seq[i]); s->seq[i] = NULL; } b->num_correct_base += align; // uint64_t k; // b->num_read_base += overlap_statistics(&b->olist, NULL, NULL, 1); // for (k = 0; k < bl->tk.n; k++) { // if(bl->tk.a[k].sec == 0) b->num_correct_base += bl->tk.a[k].qe - bl->tk.a[k].qs; // if(bl->tk.a[k].sec > 0) b->num_recorrect_base += bl->tk.a[k].qe - bl->tk.a[k].qs; // } // for (k = 0; k < bl->lo.n; k++) { // b->num_read_base += bl->lo.a[k].qe - bl->lo.a[k].qs; // } // uint32_t l1 = overlap_statistics(&b->olist, s->uu->ug, 1), l2 = overlap_statistics(&b->olist, s->uu->ug, 2); // // if(l1 == 0 && l2 > 0) fprintf(stderr, "[M::%s::%lu::no_match]\n", UL_INF.nid.a[s->id+i].a, s->len[i]); // fprintf(stderr, "[M::%s::%lu::] l1->%u; l2->%u\n", UL_INF.nid.a[s->id+i].a, s->len[i], l1, l2); if(km) { destory_overlap_region_alloc_buf(km, &b->olist, 1); destory_Correct_dumy_buf(km, &b->correct, 1); destoryHaplotypeEvdience_buf(km, &b->hap, 1); } } void dump_gaf(mg_gres_a *hits, const mg_gchains_t *gs, uint32_t only_p) { if (gs == NULL || gs->n_gc == 0 || gs->n_lc == 0) return; uint64_t i, j; int64_t q_span; mg_gres_t *p = NULL; kv_pushp(mg_gres_t, *hits, &p); memset(p, 0, sizeof(*p)); p->n_gc = 0; p->n_lc = 0; p->qid = gs->qid; p->qlen = gs->qlen; // p->n_gc = gs->n_gc; p->n_lc = gs->n_lc; p->qid = gs->qid; p->qlen = gs->qlen; // MALLOC(p->gc, p->n_gc); memcpy(p->gc, gs->gc, p->n_gc); for (i = 0; i < (uint64_t)gs->n_gc; ++i) { const mg_gchain_t *t = &gs->gc[i];///one of the gchain if(only_p && t->id != t->parent) continue; if (t->cnt == 0) continue; p->n_gc++; p->n_lc += t->cnt; } if (p->n_gc == 0) { hits->n--; return; } MALLOC(p->gc, p->n_gc); MALLOC(p->lc, p->n_lc); p->n_gc = p->n_lc = 0; for (i = 0; i < (uint64_t)gs->n_gc; ++i) { const mg_gchain_t *t = &gs->gc[i];///one of the gchain if(only_p && t->id != t->parent) continue; if (t->cnt == 0) continue; p->gc[p->n_gc] = *t; p->gc[p->n_gc].off = p->n_lc; for (j = 0; j < (uint64_t)t->cnt; ++j) { const mg_llchain_t *q = &gs->lc[t->off + j]; p->lc[p->n_lc+j].cnt = q->cnt; p->lc[p->n_lc+j].score = q->score; p->lc[p->n_lc+j].v = q->v; if(q->cnt) { q_span = (int32_t)(gs->a[q->off].y>>32&0xff); p->lc[p->n_lc+j].qs = (int32_t)gs->a[q->off].y + 1 - q_span;///calculated by the first lchain p->lc[p->n_lc+j].ts = (int32_t)gs->a[q->off].x + 1 - q_span;///calculated by the first lchain p->lc[p->n_lc+j].qe = (int32_t)gs->a[q->off + q->cnt - 1].y + 1; p->lc[p->n_lc+j].te = (int32_t)gs->a[q->off + q->cnt - 1].x + 1; } else { p->lc[p->n_lc+j].qs = p->lc[p->n_lc+j].qe = p->lc[p->n_lc+j].ts = p->lc[p->n_lc+j].te = (uint32_t)-1; } // mg_sprintf_lite(s, "%c%s", "><"[q->v&1], g->seg[q->v>>1].name); } p->n_gc++; p->n_lc += t->cnt; } } static void *worker_ul_pipeline(void *data, int step, void *in) // callback for kt_pipeline() { uldat_t *p = (uldat_t*)data; ///uint64_t total_base = 0, total_pair = 0; if (step == 0) { // step 1: read a block of sequences int ret; uint64_t l; utepdat_t *s; CALLOC(s, 1); s->ha_flt_tab = p->ha_flt_tab; s->ha_idx = p->ha_idx; s->id = p->total_pair; s->opt = p->opt; s->ug = p->ug; s->uopt = p->uopt; s->rg = p->rg; while ((ret = kseq_read(p->ks)) >= 0) { if (p->ks->seq.l < (uint64_t)p->opt->k) continue; if (s->n == s->m) { s->m = s->m < 16? 16 : s->m + (s->n>>1); REALLOC(s->len, s->m); REALLOC(s->seq, s->m); } /**if(asm_opt.flag & HA_F_VERBOSE_GFA)**/ { kv_push(uint64_t, p->nn, p->ks->name.l+p->nn.tl); kv_resize(char, p->nn.cc, p->ks->name.l+p->nn.tl); memcpy(p->nn.cc.a+p->nn.tl, p->ks->name.s, p->ks->name.l); p->nn.tl += p->ks->name.l; } l = p->ks->seq.l; MALLOC(s->seq[s->n], l); s->sum_len += l; memcpy(s->seq[s->n], p->ks->seq.s, l); s->len[s->n++] = l; if (s->sum_len >= p->chunk_size) break; } p->total_pair += s->n; if (s->sum_len == 0) free(s); else return s; } else if (step == 1) { // step 2: alignment uint64_t i; utepdat_t *s = (utepdat_t*)in; CALLOC(s->mzs, p->n_thread); CALLOC(s->sps, p->n_thread); CALLOC(s->gcs, s->n); s->buf = (mg_tbuf_t**)calloc(p->n_thread, sizeof(mg_tbuf_t*)); for (i = 0; i < p->n_thread; ++i) s->buf[i] = mg_tbuf_init(); kt_for(p->n_thread, worker_for_ul_alignment, s, s->n); for (i = 0; i < (uint64_t)s->n; ++i) { free(s->seq[i]); p->total_base += s->len[i]; } free(s->seq); free(s->len); for (i = 0; i < p->n_thread; ++i) { mg_tbuf_destroy(s->buf[i]); free(s->mzs[i].a); free(s->sps[i].a); } free(s->buf); free(s->mzs); free(s->sps); return s; } else if (step == 2) { // step 3: dump utepdat_t *s = (utepdat_t*)in; uint64_t i; for (i = 0; i < (uint64_t)s->n; ++i) { // if(s->pos[i].s == (uint64_t)-1) continue; // kv_push(pe_hit, p->hits.a, s->pos[i]); if(!s->gcs[i]) continue; dump_gaf(&(p->hits), s->gcs[i], 1); free(s->gcs[i]->gc); free(s->gcs[i]->a); free(s->gcs[i]->lc); free(s->gcs[i]); } free(s->gcs); free(s); } return 0; } int alignment_ul_pipeline(uldat_t* sl, const enzyme *fn) { double index_time = yak_realtime(); int i; for (i = 0; i < fn->n; i++){ gzFile fp; if ((fp = gzopen(fn->a[i], "r")) == 0) return 0; sl->ks = kseq_init(fp); kt_pipeline(3, worker_ul_pipeline, sl, 3); kseq_destroy(sl->ks); gzclose(fp); } sl->hits.total_base = sl->total_base; sl->hits.total_pair = sl->total_pair; fprintf(stderr, "[M::%s::%.3f] ==> Qualification\n", __func__, yak_realtime()-index_time); return 1; } void push_uc_block_t(kv_ul_ov_t *z, char **seq, uint64_t *len, uint64_t b_id) { uint64_t k, l, rid; for (k = 1, l = 0; k <= z->n; k++) { if(k == z->n || z->a[k].qn != z->a[l].qn) { rid = b_id + z->a[l].qn; append_ul_t(&UL_INF, &rid, NULL, 0, seq[z->a[l].qn], len[z->a[l].qn], z->a + l, k - l, P_CHAIN_COV); l = k; } } } static void *worker_ul_scall_pipeline(void *data, int step, void *in) // callback for kt_pipeline() { uldat_t *p = (uldat_t*)data; ///uint64_t total_base = 0, total_pair = 0; if (step == 0) { // step 1: read a block of sequences int ret; uint64_t l; utepdat_t *s; CALLOC(s, 1); s->ha_flt_tab = p->ha_flt_tab; s->ha_idx = p->ha_idx; s->id = p->total_pair; s->opt = p->opt; s->uu = p->uu; s->uopt = p->uopt; s->rg = p->rg; while ((ret = kseq_read(p->ks)) >= 0) { if (p->ks->seq.l < (uint64_t)p->opt->k) continue; if (s->n == s->m) { s->m = s->m < 16? 16 : s->m + (s->n>>1); REALLOC(s->len, s->m); REALLOC(s->seq, s->m); } append_ul_t(&UL_INF, NULL, p->ks->name.s, p->ks->name.l, NULL, 0, NULL, 0, P_CHAIN_COV); l = p->ks->seq.l; MALLOC(s->seq[s->n], l); s->sum_len += l; memcpy(s->seq[s->n], p->ks->seq.s, l); s->len[s->n++] = l; if (s->sum_len >= p->chunk_size) break; } p->total_pair += s->n; if (s->sum_len == 0) free(s); else return s; } else if (step == 1) { // step 2: alignment utepdat_t *s = (utepdat_t*)in; uint64_t i; CALLOC(s->hab, p->n_thread); CALLOC(s->ll, p->n_thread); // CALLOC(s->buf, p->n_thread); for (i = 0; i < p->n_thread; ++i) { // s->buf[i] = mg_tbuf_init(); // s->hab[i] = ha_ovec_buf_init(s->buf[i]->km, 0, 0, 1); // s->buf[i] = NULL; // s->hab[i] = ha_ovec_buf_init(NULL, 0, 0, 1); s->hab[i] = ha_ovec_init(0, 0, 1); } kt_for(p->n_thread, worker_for_ul_scall_alignment, s, s->n); ///debug /** uint64_t i; CALLOC(s->mzs, p->n_thread); CALLOC(s->sps, p->n_thread); CALLOC(s->gcs, s->n); s->buf = (mg_tbuf_t**)calloc(p->n_thread, sizeof(mg_tbuf_t*)); for (i = 0; i < p->n_thread; ++i) s->buf[i] = mg_tbuf_init(); kt_for(p->n_thread, worker_for_ul_alignment, s, s->n); for (i = 0; i < (uint64_t)s->n; ++i) { free(s->seq[i]); p->total_base += s->len[i]; } free(s->seq); free(s->len); for (i = 0; i < p->n_thread; ++i) { mg_tbuf_destroy(s->buf[i]); free(s->mzs[i].a); free(s->sps[i].a); } **/ for (i = 0; i < p->n_thread; ++i) { s->num_bases += s->hab[i]->num_read_base; s->num_corrected_bases += s->hab[i]->num_correct_base; s->num_recorrected_bases += s->hab[i]->num_recorrect_base; // mg_tbuf_destroy(s->buf[i]); ha_ovec_destroy(s->hab[i]); free(s->ll[i].lo.a); /**free(s->ll[i].tk.a);**/ free(s->ll[i].srt.a.a); } free(s->hab); /**free(s->ll);**/ // free(s->buf); //free(s->mzs); free(s->sps); return s; } else if (step == 2) { // step 3: dump utepdat_t *s = (utepdat_t*)in; uint64_t i, rid; p->num_bases += s->num_bases; p->num_corrected_bases += s->num_corrected_bases; p->num_recorrected_bases += s->num_recorrected_bases; for (i = 0; i < p->n_thread; ++i) { push_uc_block_t(&(s->ll[i].tk), s->seq, s->len, s->id); free(s->ll[i].tk.a); } for (i = 0; i < (uint64_t)s->n; ++i) { rid = s->id + i; if(UL_INF.n > rid && UL_INF.a[rid].rlen != s->len[i]) { append_ul_t(&UL_INF, &rid, NULL, 0, s->seq[i], s->len[i], NULL, 0, P_CHAIN_COV); } free(s->seq[i]); } /** for (i = 0; i < (uint64_t)s->n; ++i) { ///debug // if(s->pos[i].s == (uint64_t)-1) continue; // kv_push(pe_hit, p->hits.a, s->pos[i]); // if(!s->gcs[i]) continue; // dump_gaf(&(p->hits), s->gcs[i], 1); // free(s->gcs[i]->gc); free(s->gcs[i]->a); free(s->gcs[i]->lc); free(s->gcs[i]); rid = s->id + i; append_ul_t(&UL_INF, &rid, NULL, 0, s->seq[i], s->len[i], NULL, 0, P_CHAIN_COV); // fprintf(stderr, "%.*s\n", (int)s->len[i], s->seq[i]); free(s->seq[i]); p->total_base += s->len[i]; } **/ ///debug /** free(s->gcs); **/ free(s->ll); free(s->len); free(s->seq); free(s); } return 0; } utg_rid_dt *get_r_ug_region(utg_rid_t *idx, uint64_t *n, uint64_t rid) { (*n) = idx->idx[rid+1] - idx->idx[rid]; return (*n)?idx->p.a + idx->idx[rid]:NULL; } void rov2uov(uint64_t rid, const ul_idx_t *uref, utg_rid_dt *ru_map, uc_block_t *rovlp, ul_ov_t *res, uint32_t adjust_rev) { uint64_t ori = ru_map->u&1, ts, te; if(!ori) { ts = rovlp->ts; te = rovlp->te; } else { ts = uref->r_ug->rg->seq[rid].len - rovlp->te; te = uref->r_ug->rg->seq[rid].len - rovlp->ts; } ts += ru_map->off; te += ru_map->off; memset(res, 0, sizeof(*res)); res->qn = 0; res->qs = rovlp->qs; res->qe = rovlp->qe; res->tn = ru_map->u>>1; res->ts = ts; res->te = te; res->el = rovlp->el; res->rev = (rovlp->rev == ori?0:1); if(adjust_rev && res->rev) {///for linear chaining res->ts = uref->ug->g->seq[res->tn].len - te; res->te = uref->ug->g->seq[res->tn].len - ts; } } void print_ul_ov_t(ul_ov_t *xs, const char* cmd) { fprintf(stderr, "%s\t%s\t%u\t%u\t%c\t%.*s\t%u\t%u\n", cmd, UL_INF.nid.a[xs->qn].a, xs->qs, xs->qe, "+-"[xs->rev], (int)Get_NAME_LENGTH(R_INF, ((xs->tn<<1)>>1)), Get_NAME(R_INF, ((xs->tn<<1)>>1)), xs->ts, xs->te); } void gl_rg2ug_gen(ul_vec_t *r_cl, kv_ul_ov_t *u_cl, const ul_idx_t *uref, uint64_t is_el) { uint64_t k, a_k, a_n; uc_block_t *z; utg_rid_dt *a; ul_ov_t *p; u_cl->n = 0; for (k = 0; k < r_cl->bb.n; k++) { z = &(r_cl->bb.a[k]); if(z->base) continue; if(is_el && (!(z->el))) continue; a = get_r_ug_region(uref->r_ug, &a_n, z->hid); if(!a) continue; for (a_k = 0; a_k < a_n; a_k++) { kv_pushp(ul_ov_t, *u_cl, &p); // fprintf(stderr, "\n+[M::%s::] %u\t%u\t%c\t%.*s(%u)\t%u\t%u\n", __func__, z->qs, z->qe, "+-"[z->rev], // (int)Get_NAME_LENGTH(R_INF, z->hid), Get_NAME(R_INF, z->hid), (uint32_t)Get_READ_LENGTH(R_INF, z->hid), z->ts, z->te); // fprintf(stderr, "*[M::%s::] utg%.6d%c(%u)\t%c\t%u\n", __func__, // (int32_t)(a[a_k].u>>1)+1, "lc"[uref->ug->u.a[a[a_k].u>>1].circ], uref->ug->u.a[a[a_k].u>>1].len, // "+-"[a[a_k].u&1], a[a_k].off); rov2uov(z->hid, uref, &(a[a_k]), z, p, 1); p->tn <<= 1; p->tn |= p->rev; p->qn = uref->r_ug->idx[z->hid] + a_k;//for linear chain // fprintf(stderr, "[M::%s::id->%ld] idx->n:%lu\n", __func__, ulid, (uint64_t)idx->n); // fprintf(stderr, "-[M::%s::] %u\t%u\t%c\tutg%.6d%c(%u)\t%u\t%u\n", __func__, p->qs, p->qe, "+-"[p->rev], // (int32_t)(p->tn>>1)+1, "lc"[uref->ug->u.a[p->tn>>1].circ], uref->ug->u.a[p->tn>>1].len, p->ts, p->te); } } } void adjust_rev_tse(ul_ov_t *x, int64_t tlen, int64_t *ts, int64_t *te) { *ts = x->ts; *te = x->te; if(x->rev) { *ts = tlen - x->te; *te = tlen - x->ts; } } uint64_t get_add_cov_score(const ul_idx_t *uref, int64_t ps, int64_t pe, int64_t cs, int64_t ce, int64_t uid, int64_t *cov_i) { int64_t os = MAX(ps, cs), oe = MIN(pe, ce); int64_t ovlp = ((oe > os)? (oe - os):0); // fprintf(stderr, "ovlp:%ld, os:%ld, oe:%ld, ps:%ld, pe:%ld, cs:%ld, ce::%ld\n", ovlp, os, oe, ps, pe, cs, ce); if(ovlp > 0) { return (os>cs?retrieve_u_cov_region(uref, uid, 0, cs, os, cov_i):0) + (ce>oe?retrieve_u_cov_region(uref, uid, 0, oe, ce, cov_i):0); } return retrieve_u_cov_region(uref, uid, 0, cs, ce, cov_i); } uint64_t linear_chain_dp(ul_ov_t *ch, int64_t ch_n, ul_ov_t *sv, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, uint64_t *idx, uint64_t *track, ma_ug_t *ug, int64_t chain_offset) { ///all in[].el must be 1 if(ch_n == 0) return 0; int64_t /**mm_ovlp, x,**/ i, j, k, sc, csc, mm_sc, mm_idx, its, ite, jts, jte, cov_i, dq, dt, dd, mm; ul_ov_t *li = NULL, *lj = NULL; radix_sort_ul_ov_srt_qe(ch, ch + ch_n); for (i = 1, j = 0; i <= ch_n; i++) { // if(i < ch_n) { // li = &(ch[i]); // fprintf(stderr, "##(%ld) %u\t%u\t%c\tutg%.6d%c(%u)\t%u\t%u\tmm_idx:%ld\tmm_sc:%ld\n", i, li->qs, li->qe, "+-"[li->rev], // (int32_t)(li->tn)+1, "lc"[uref->ug->u.a[li->tn].circ], uref->ug->u.a[li->tn].len, li->ts, li->te, mm_idx, mm_sc); // } if (i == ch_n || ch[i].qe != ch[j].qe) { if(i - j > 1) { radix_sort_ul_ov_srt_qs(ch+j, ch+i); } j = i; } } // fprintf(stderr, "[M::%s::] ch_n:%ld\n", __func__, ch_n); for (i = 0; i < ch_n; ++i) { li = &(ch[i]); // mm_ovlp = max_ovlp_src(uopt, ((li->tn<<1)|li->rev)^1); // x = (li->qs + mm_ovlp)*diff_ec_ul; // if(x < bw) x = bw; // x += li->qs + mm_ovlp; // if (x > qlen+1) x = qlen+1; // x = find_ul_ov_max(i, ch, x+G_CHAIN_INDEL); adjust_rev_tse(li, ug->g->seq[li->tn].len, &its, &ite); cov_i = 0; csc = retrieve_u_cov_region(uref, li->tn, 0, its, ite, &cov_i); mm_sc = csc; mm_idx = -1; for (j = i-1/**x**/; j >= 0; --j) { lj = &(ch[j]); // fprintf(stderr, "<0>\n"); if(lj->qs <= li->qs && lj->qe <= li->qe && lj->ts <= li->ts && lj->te <= li->te) {///co-linear assert(li->tn == lj->tn && li->rev == lj->rev); // fprintf(stderr, "<1>\n"); if(lj->qs == li->qs && lj->qe == li->qe && lj->ts == li->ts && lj->te == li->te) continue; dq = li->qe - lj->qs; dt = li->te - lj->ts; dd = (dq>dt? dq-dt:dt-dq); mm = MAX(dq, dt); mm *= diff_ec_ul; if(mm < bw) mm = bw; // fprintf(stderr, "+++i->%ld, j->%ld, dd->%ld, mm->%ld\n", i, j, dd, mm); if(dd <= mm) {///pass distance checking adjust_rev_tse(lj, ug->g->seq[lj->tn].len, &jts, &jte); sc = get_add_cov_score(uref, jts, jte, its, ite, li->tn, &cov_i) + pop_sc(track[j]); if((sc > mm_sc) || (sc == mm_sc && mm_idx == -1)) { ///must be >= instead of > mm_sc = sc, mm_idx = j; } // fprintf(stderr, "%ld, its:%ld, ite:%ld>, %ld, jts:%ld, jte:%ld> sc:%ld, pop_sc(track[j]):%ld, csc:%ld\n", // i, its, ite, j, jts, jte, sc, pop_sc(track[j]), csc); } } } // fprintf(stderr, "##(%ld) %u\t%u\t%c\tutg%.6d%c(%u)\t%u\t%u\tmm_idx:%ld\tmm_sc:%ld\n", i, li->qs, li->qe, "+-"[li->rev], // (int32_t)(li->tn)+1, "lc"[uref->ug->u.a[li->tn].circ], uref->ug->u.a[li->tn].len, li->ts, li->te, mm_idx, mm_sc); track[i] = push_sc_pre(mm_sc, mm_idx); li->sec = (mm_idx<0?0x3FFFFFFF:i-mm_idx); sv[i] = *li; } int64_t n_u; for (k = ch_n-1, n_u = 0; k >= 0; --k) { if(track[k]&((uint64_t)0x80000000)) continue; i = k; ch[n_u]=sv[i]; sc = pop_sc(track[i]); for (;i>=0;) { track[i] |= ((uint64_t)0x80000000); if(sv[i].qs < ch[n_u].qs) ch[n_u].qs = sv[i].qs; if(sv[i].ts < ch[n_u].ts) ch[n_u].ts = sv[i].ts; if(sv[i].qe > ch[n_u].qe) ch[n_u].qe = sv[i].qe; if(sv[i].te > ch[n_u].te) ch[n_u].te = sv[i].te; ch[n_u].qn = i;//start idx of read alignment in chain i = pop_pre(track[i]); } adjust_rev_tse(&(ch[n_u]), ug->g->seq[ch[n_u].tn].len, &its, &ite); ch[n_u].ts = its; ch[n_u].te = ite; ch[n_u].sec = (sc>0x3FFFFFFF?0x3FFFFFFF:sc); ch[n_u].qn += chain_offset; //start idx of read alignment in chain ch[n_u].tn = k + chain_offset; //end idx of read alignment in chain n_u++; } for (i = 0; i < ch_n; ++i) { adjust_rev_tse(&(sv[i]), ug->g->seq[sv[i].tn].len, &its, &ite); sv[i].ts = its; sv[i].te = ite; k = pop_pre(track[i]); sv[i].qn = k>=0?k+chain_offset:(uint32_t)-1; } return n_u; } void gen_linear_chains(kv_ul_ov_t *res, kv_ul_ov_t *buf, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, glchain_t *bufg, st_mt_t *bufs) { uint64_t k, l, z, an, m; radix_sort_ul_ov_srt_tn(res->a, res->a + res->n); kv_resize(ul_ov_t, *buf, res->n); buf->n = res->n; for (k = 1, l = m = 0; k <= res->n; k++) { if(k == res->n || res->a[k].tn != res->a[l].tn) {///qn <- (tn|rev) kv_resize(uint64_t, bufg->srt.a, k-l); kv_resize(uint64_t, *bufs, k-l); for (z = l; z < k; z++) res->a[z].tn>>=1; // fprintf(stderr, "\n*[M::%s::] %c\tutg%.6d%c(%u)\tocc:[%lu, %lu)\n", __func__, "+-"[res->a[l].rev], (int32_t)(res->a[l].tn)+1, // "lc"[uref->ug->u.a[res->a[l].tn].circ], uref->ug->u.a[res->a[l].tn].len, l, k); an = l + linear_chain_dp(res->a+l, k-l, buf->a+l, uref, uopt, bw, diff_ec_ul, qlen, max_skip, bufg->srt.a.a, bufs->a, uref->ug, l); for (z = l; z < an; z++) res->a[m++] = res->a[z]; // fprintf(stderr, "#occ:[%lu, %lu)\n", l, an); l = k; } } res->n = m; } void gen_end_coord(ul_ov_t *z, int64_t qlen, int64_t tlen, int64_t *r_qs, int64_t *r_qe, int64_t *r_ts, int64_t *r_te) { int64_t qs, qe, ts, te, qtail, ttail; qs = z->qs; qe = z->qe; ts = z->ts; te = z->te; if(z->rev) { ts = tlen - z->te; te = tlen - z->ts; } if(qs <= ts) { ts -= qs; qs = 0; } else { qs -= ts; ts = 0; } qtail = qlen - qe; ttail = tlen - te; if(qtail <= ttail) { qe = qlen; te += qtail; } else { te = tlen; qe += ttail; } if(r_qs) (*r_qs) = qs; if(r_qe) (*r_qe) = qe; if(r_ts) (*r_ts) = ts; if(r_te) (*r_te) = te; if(z->rev) { if(r_ts) (*r_ts) = tlen - te; if(r_te) (*r_te) = tlen - ts; } } uint32_t is_end_check(uint32_t v, ul_ov_t *z, asg_t *g) { if(v&1) { if(z->ts==0) return 1; } else { if(z->te==g->seq[v>>1].len) return 1; } return 0; } int64_t simple_g_chain_dp(kv_ul_ov_t *in, ul_ov_t *buf, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t qlen, int64_t max_skip, uint64_t *srt, uint64_t *idx, uint64_t *track) { if(in->n == 0) return 0; uint32_t ai_v, aj_v, rev_n; ma_ug_t *ug = uref->ug; int64_t mm_ovlp, x, i, j, k, sc, csc, mm_sc, mm_idx, qo, share, in_n = in->n; int64_t iqs, iqe, its, ite, i_end, j_end; ul_ov_t *ai, *aj, *e_ai, *e_aj, rev_t; for (i = 0; i < in_n; i++) { gen_end_coord(&(in->a[i]), qlen, ug->u.a[in->a[i].tn].len, NULL, &iqe, NULL, NULL); srt[i] = iqe; srt[i] <<= 32; srt[i] |= (uint64_t)i; } radix_sort_gfa64(srt, srt+in_n); for (i = 0; i < in_n; i++) buf[i] = in->a[(uint32_t)srt[i]]; memcpy(in->a, buf, in_n *sizeof((*buf)));///all alignments have been sorted by the real end-qe for (i = 0; i < in_n; ++i) { ai = &(in->a[i]); ai_v = (ai->tn<<1)|ai->rev; e_ai = &(buf[i]); i_end = 0; gen_end_coord(ai, qlen, ug->u.a[ai->tn].len, &iqs, &iqe, &its, &ite); e_ai->qs = iqs; e_ai->qe = iqe; e_ai->ts = its; e_ai->te = ite; mm_ovlp = max_ovlp(uref->ug->g, ai_v^1); x = (e_ai->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += e_ai->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_ul_ov_max(i, buf, x+G_CHAIN_INDEL); i_end = is_end_check(ai_v^1, ai, uref->ug->g); csc = mm_sc = e_ai->sec; mm_idx = -1; for (j = x; j >= 0; --j) { // collect potential destination vertices aj = &(in->a[j]); aj_v = (aj->tn<<1)|aj->rev; e_aj = &(buf[i]); j_end = 0; if(e_aj->qe+G_CHAIN_INDEL <= e_ai->qs) break;//even this pair has a overlap, its length will be very small; just ignore if(e_aj->qs >= e_ai->qs+G_CHAIN_INDEL) continue; // lj is contained in li on the query coordinate; 128 for indel offset qo = infer_rovlp(e_ai, e_aj, NULL, NULL, NULL, ug); ///overlap length in query (UL read) if(ai_v != aj_v && get_ecov_adv(uref, uopt, ai_v^1, aj_v^1, bw, diff_ec_ul, qo, 0, &share)) { sc = csc + pop_sc(track[j]); j_end = is_end_check(aj_v, aj, uref->ug->g); if(i_end && j_end) sc -= (share>=csc?csc:share); if(sc > mm_sc) mm_sc = sc, mm_idx = j; } } track[i] = push_sc_pre(mm_sc, mm_idx); srt[i] = track[i]>>32; srt[i] <<= 32; srt[i] |= i; } int64_t n_v, n_u, n_v0; radix_sort_gfa64(srt, srt+in_n); for (k = in_n-1, n_v = n_u = 0; k >= 0; --k) { n_v0 = n_v; for (i = (uint32_t)srt[k]; i >= 0 && (track[i]&((uint64_t)0x80000000)) == 0;){ buf[n_v] = in->a[i]; gen_end_coord(&(buf[n_v]), qlen, ug->u.a[buf[n_v].tn].len, &iqs, &iqe, &its, &ite); buf[n_v].qs = iqs; buf[n_v].qe = iqe; buf[n_v].ts = its; buf[n_v].te = ite; track[i] |= ((uint64_t)0x80000000); i = pop_pre(track[i]); n_v++; } if(n_v0 == n_v) continue; sc = (i<0?(pop_sc(srt[k])):(pop_sc(srt[k])-pop_sc(track[i]))); idx[n_u++] = ((uint64_t)sc<<32)|(n_v-n_v0); } for (k = 0, n_v = n_v0 = 0; k < n_u; k++) { n_v0 = n_v; n_v += (uint32_t)idx[k]; in->a[k].qn = idx[k]>>32;//score in->a[k].ts = n_v0; in->a[k].te = n_v;///idx rev_n = ((uint32_t)idx[k])>>1; ///we need to consider contained reads; so determining qs is not such easy in->a[k].qs = (uint32_t)-1; in->a[k].qe = buf[n_v0].qe; for (i = 0; i < rev_n; i++) { rev_t = buf[n_v0+i]; buf[n_v0+i] = buf[n_v-i-1]; buf[n_v-i-1] = rev_t; if(in->a[k].qs > buf[n_v0+i].qs) in->a[k].qs = buf[n_v0+i].qs; if(in->a[k].qs > buf[n_v-i-1].qs) in->a[k].qs = buf[n_v-i-1].qs; } if(((uint32_t)idx[k])&1) { if(in->a[k].qs > buf[n_v0+i].qs) in->a[k].qs = buf[n_v0+i].qs; } // fprintf(stderr, "[M::%s] k:%ld, qs:%u, qe:%u, chain_occ:%u, chain_score:%u\n", __func__, k, // res->a[k].qs, res->a[k].qe, res->a[k].te - res->a[k].ts, res->a[k].qn); } in->n = n_u; radix_sort_ul_ov_srt_qn(in->a, in->a + in->n);//sort by score return n_v; } /** uint32_t uov2rov(const ul_idx_t *uref, ul_ov_t *r_al, ul_ov_t *ul_al, ul_ov_t *res) { int64_t y_s, y_e, y_bs, y_be, x_s, x_e, q_s, q_e, s_shift, e_shift; y_s = MAX(r_al->ts, ul_al->ts); y_e = MIN(r_al->te, ul_al->te); if(y_s > y_e) return 0; res->tn = r_al->qn; res->ts = y_s; res->te = y_e; res->el = 1; res->rev = r_al->rev; res->sec = 0; s_shift = get_offset_adjust(y_s-r_al->ts, r_al->te-r_al->ts, r_al->qe-r_al->qs); e_shift = get_offset_adjust(r_al->te-y_e, r_al->te-r_al->ts, r_al->qe-r_al->qs); if(r_al->rev) { y_s = s_shift; s_shift = e_shift; e_shift = y_s; } res->qn = 0; res->qs = r_al->qs+s_shift; res->qe = r_al->qe-e_shift; return 1; } void update_ul_vec_t() { } void ug2rg_gen(ul_ov_t *a, int64_t an, ul_vec_t *qn, const ul_idx_t *uref, ul_vec_t *rch) { ul_ov_t *ot, p, res; uint64_t i, l, m; ma_utg_t *u; uc_block_t *b; int64_t z, ff, iqs, iqe, its, ite; for (z = 0; z < an; z++) { gen_end_coord(&(a[z]), rch->rlen, uref->ug->u.a[a[z].tn].len, &iqs, &iqe, NULL, NULL); o = &(a[z]); u = &(uref->ug->u.a[o->tn]); for (i = l = 0; i < u->n; i++) { p.tn = o->tn; p.rev = (u->a[i]>>32)&1; p.qn = u->a[i]>>33;///tn is unitig, qn is HiFi read p.qs = 0; p.qe = Get_READ_LENGTH(R_INF, (u->a[i]>>33)); p.ts = l; p.te = l + Get_READ_LENGTH(R_INF, (u->a[i]>>33)); l += (uint32_t)u->a[i]; if(p.te <= o->ts) continue; if(p.ts >= o->te) break; ff = uov2rov(uref, &p, o, &res); assert(ff); if(ff) { kv_pushp(uc_block_t, rch->bb, &b); b->hid = res.tn; b->rev = res.rev; b->base = 0; b->el = res.el; b->pchain = 1; b->qs = res.qs; b->qe = res.qe; b->ts = res.ts; b->te = res.te; } } } } **/ void extend_end_coord(mg_lchain_t *li, ul_ov_t *ui, const int64_t qlen, const int64_t rlen, int64_t *r_qs, int64_t *r_qe, int64_t *r_rs, int64_t *r_re) { int64_t qs = 0, qe = 0, rs = 0, re = 0, rev = 0, qtail = 0, rtail = 0; if(li) { qs = li->qs; qe = li->qe; rs = li->rs; re = li->re; rev = li->v&1; if(rev) { rs = rlen - li->re; re = rlen - li->rs; } } if(ui) { qs = ui->qs; qe = ui->qe; rs = ui->ts; re = ui->te; rev = ui->rev; if(rev) { rs = rlen - ui->te; re = rlen - ui->ts; } } if(qs <= rs) { rs -= qs; qs = 0; } else { qs -= rs; rs = 0; } qtail = qlen - qe; rtail = rlen - re; if(qtail <= rtail) { qe = qlen; re += qtail; } else { re = rlen; qe += rtail; } if(r_qs) (*r_qs) = qs; if(r_qe) (*r_qe) = qe; if(r_rs) (*r_rs) = rs; if(r_re) (*r_re) = re; if(rev) { if(r_rs) (*r_rs) = rlen - re; if(r_re) (*r_re) = rlen - rs; } } void dump_linear_chain(asg_t *g, kv_ul_ov_t *lidx, kv_ul_ov_t *autom, vec_mg_lchain_t *res, int64_t qlen) { uint64_t k; int64_t iqs, iqe, its, ite; res->n = 0; kv_resize(mg_lchain_t, *res, lidx->n); res->n = lidx->n; for (k = 0; k < lidx->n; k++) { memset(&(res->a[k]), 0, sizeof(res->a[k])); res->a[k].v = (autom->a[lidx->a[k].tn].tn<<1)|lidx->a[k].rev; ///.off -> idx of original chain; cnt -> score of the chain res->a[k].off = k; res->a[k].score = lidx->a[k].sec; res->a[k].qs = lidx->a[k].qs; res->a[k].qe = lidx->a[k].qe; res->a[k].rs = lidx->a[k].ts; res->a[k].re = lidx->a[k].te; extend_end_coord(&(res->a[k]), NULL, qlen, g->seq[res->a[k].v>>1].len, &iqs, &iqe, &its, &ite); res->a[k].qs = iqs; res->a[k].qe = iqe; res->a[k].rs = its; res->a[k].re = ite; // fprintf(stderr, "chain_id:%d\t%u\t%u\t%c\tutg%.6dl(%u)\t%u\t%u\n", // res->a[k].off, res->a[k].qs, res->a[k].qe, "+-"[res->a[k].v&1], (int32_t)(res->a[k].v>>1)+1, // g->seq[res->a[k].v>>1].len, res->a[k].rs, res->a[k].re); } } int64_t find_mg_lchain_max(int64_t n, const mg_lchain_t *a, int32_t x) { int64_t s = 0, e = n; if (n == 0) return -1; if (a[n-1].qe < x) return n - 1; if (a[0].qe >= x) return -1; while (e > s) { // TODO: finish this block int64_t m = s + (e - s) / 2; if (a[m].qe >= x) e = m; else s = m + 1; } assert(s == e); return s; } int64_t hc_target_len(asg_t *g, mg_lchain_t *s, mg_lchain_t *e) { // int64_t ql = s->qe - e->qe, tp, tm; // if((s->v^1)&1) tp = g->seq[s->v>>1].len - s->re; // else tp = s->rs; // if((e->v^1)&1) tm = g->seq[e->v>>1].len - e->re; // else tm = e->rs; int64_t ql = (int64_t)s->qs - (int64_t)e->qe, tp, tm; int64_t sts, ete; sts = (s->v&1)?g->seq[s->v>>1].len-s->re:s->rs; tp = g->seq[s->v>>1].len - sts; ete = (e->v&1)?g->seq[e->v>>1].len-e->rs:e->re; tm = g->seq[e->v>>1].len - ete; // fprintf(stderr, "[M::%s::] ql:%ld, tp:%ld, tm:%ld, sts:%ld, ete:%ld\n", __func__, ql, tp, tm, sts, ete); return ql + tp - tm; } inline int32_t cal_gchain_sc(const mg_path_dst_t *dj, const mg_lchain_t *li, const mg_lchain_t *lc, int64_t *f, int64_t b_w, float diff_thre, float chn_pen_gap) { // const mg_lchain_t *lj; int32_t gap, sc; float lin_pen, log_pen; if (dj->n_path == 0) return INT32_MIN; gap = dj->dist - dj->target_dist; // lj = &lc[dj->meta]; if (gap < 0) gap = -gap; if ((gap > ((dj->target_dist)*diff_thre)) && (gap > b_w)) return INT32_MIN; // if (lj->qe <= li->qs) sc = li->score; // else sc = (int32_t)((double)(li->qe - lj->qe) / (li->qe - li->qs) * li->score + .499); // dealing with overlap on query sc = li->score; //sc += dj->mlen; // TODO: is this line the right thing to do? // if (dj->is_0) sc += ref_bonus; lin_pen = chn_pen_gap * (float)gap; log_pen = gap >= 2? mg_log2(gap) : 0.0f; sc -= (int32_t)(lin_pen + log_pen); sc += f[dj->meta]; return sc; } ///max_dist is like the overlap length in string graph ///first_src_ban do not allow co-linear chain at the same node void hc_shortest_k(void *km0, const asg_t *g, uint32_t src, int32_t n_dst, mg_path_dst_t *dst, int32_t max_dist, int32_t max_k, st_mt_t *dst_done, uint64_t *dst_group, vec_sp_node_t *out, vec_mg_pathv_t *res, uint64_t first_src_ban) { sp_node_t *p, *root = 0; sp_topk_t *q; khash_t(sp) *h;/// khash_t(sp2) *h2;///alignment->vertice index void *km; khint_t k; int absent; int32_t i, j, n_done, n_found; uint32_t id; // if (res) res->n = 0;///for us, n_pathv = NULL if (n_dst <= 0) return;///n_dst: how many candidate vertices for (i = 0; i < n_dst; ++i) { // initialize mg_path_dst_t *t = &dst[i]; ///if src and dest are at the same ref id, there are already one path if (t->inner)///if two chains are at the same ref id t->dist = 0, t->n_path = 1, t->path_end = -1; else t->dist = -1, t->n_path = 0, t->path_end = -1; } if (max_k > MG_MAX_SHORT_K) max_k = MG_MAX_SHORT_K; km = km_init2(km0, 0x4000); // multiple dst[] may have the same dst[].v. We need to group them first. // in other words, one ref id may have multiple dst alignment chains dst_done->n = 0; kv_resize(uint64_t, *dst_done, (uint64_t)n_dst); for (i = 0; i < n_dst; ++i) { dst_group[i] = ((((uint64_t)dst[i].v)<<32)|((uint64_t)i)); dst_done->a[i] = 0; } radix_sort_gfa64(dst_group, dst_group + n_dst); h2 = kh_init2(sp2, km); // (h2+dst_group) keeps all destinations from the same ref id kh_resize(sp2, h2, n_dst * 2); ///please note that one contig in ref may have multiple alignment chains ///so h2 is a index that helps us to query it ///key(h2) = ref id; value(h2) = start_idx | occ for (i = 1, j = 0; i <= n_dst; ++i) { if (i == n_dst || dst_group[i]>>32 != dst_group[j]>>32) { k = kh_put(sp2, h2, dst_group[j]>>32, &absent); kh_val(h2, k) = (((uint64_t)j)<<32)|((uint64_t)(i-j)); assert(absent); j = i; } } h = kh_init2(sp, km); // h keeps visited vertices; path to each visited vertice kh_resize(sp, h, 16); out->n = 0; kv_resize(sp_node_t*, *out, 16); ///16 is just the initial size id = 0; p = gen_sp_node(km, src, 0, id++);///just malloc a node for src; the distance is 0 p->hash = __ac_Wang_hash(src);///hash is path hash, instead of node hash kavl_insert(sp, &root, p, 0);///should be avl tree; p is a node at avl-tree ///each cell in the hash table corresponds to one node in the graph ///each cell in the AVL tree is a path, corresponds to node in the graph k = kh_put(sp, h, src, &absent); q = &kh_val(h, k); ///for normal graph traversal, one node just has one parental node; here each node has at most 16 parental nodes q->k = 1, q->p[0] = p, q->mlen = 0, q->qs = q->qe = -1; n_done = 0; first_src_ban = first_src_ban?0:1; ///the key of avl tree: #define sp_node_cmp(a, b) (((a)->di > (b)->di) - ((a)->di < (b)->di)) ///the higher bits of (*)->di is distance to src node ///so the key of avl tree is distance ///in avl tree , one node might be saved multipe times while (kavl_size(head, root) > 0) {///thr first root is src int32_t i, nv; asg_arc_t *av; sp_node_t *r; ///note that one node in the graph (sp_node_t->v) might be visited multiple times if there are circles ///so there might be multipe cells in the avl-tree with the same (sp_node_t->v) ///delete the first cell r = kavl_erase_first(sp, &root); // take out the closest vertex in the heap (as a binary tree) //fprintf(stderr, "XX\t%d\t%d\t%d\t%c%s[%d]\t%d\n", n_out, kavl_size(head, root), n_finished, "><"[(r->v&1)^1], g->seg[r->v>>1].name, r->v, (int32_t)(r->di>>32)); ///higher 32 bits might be the distance to root node // lower 32 bits now for position in the out[] array ///r->pre keep the pre-node in the path; follow the pre it is able to recover the whole path r->di = ((r->di>>32)<<32)|((uint64_t)out->n); ///n_out is just the id in out ///so one node id in graph might be saved multiple times in avl tree and out[] kv_push(sp_node_t*, *out, r); ///r->v is the dst vertex id ///sometimes k==kh_end(h2). Some nodes are found by graph travesal but not in linear chain alignment k = kh_get(sp2, h2, r->v); // we have reached one dst vertex // note that one dst vertex may have multipe alignment chains // we can visit some nodes in graph which are not reachable during chaining // h2 is used to determine if one node is reachable or not // if(src == 2844) { // fprintf(stderr, "******src->%u, dst->%u, max_dist->%d\n", src, r->v, max_dist); // } if (k != kh_end(h2) && first_src_ban) { ///node r->v might be visited multiple times int32_t j, dist = r->di>>32, off = kh_val(h2, k) >> 32, cnt = (int32_t)kh_val(h2, k); // if(src == 2844) { // fprintf(stderr, "----src->%u, dst->%u, max_dist->%d, cnt->%d\n", src, r->v, max_dist, cnt); // } //src can reach ref id r->v; there might be not only one alignment chain in r->v //so we need to scan all of them for (j = 0; j < cnt; ++j) { mg_path_dst_t *t = &dst[(int32_t)dst_group[off + j]];///t is a linear alignment at r->v int32_t done = 0; // if((src>>1) == 51) { // fprintf(stderr, "###src->%u, dst->%u, max_dist->%d, dist:%d\n", src, r->v, max_dist, dist); // } ///the src and dest are at the same ref id, say we directly find the shortest path if (t->inner) { done = 1; } else { int32_t mlen = 0, copy = 0; ///in the first round, we just check reachability without sequence ///so h_seeds = NULL; we can assume mlen = 0 /** //path mlen = h_seeds? path_mlen(out, n_out - 1, h, t->qlen) : 0; **/ // means this alignment has never been visited before; keep it anyway // note here is the alignment, instead of node // if(src == 2844) { // fprintf(stderr, ">>src->%u, dst->%u, target_dist->%d, dist->%d, max_dist->%d\n", // src, r->v, t->target_dist, dist, max_dist); // } if (t->n_path == 0) { copy = 1; // we have a target distance; choose the closest; // there is already several paths reaching the linear alignment } else if (t->target_dist >= 0) { // we found the target path; hash is the path hash including multiple nodes, instead of node hash if (dist == t->target_dist && t->check_hash && r->hash == t->target_hash) { copy = 1, done = 1; } else { int32_t d0 = t->dist, d1 = dist; d0 = d0 > t->target_dist? d0 - t->target_dist : t->target_dist - d0; d1 = d1 > t->target_dist? d1 - t->target_dist : t->target_dist - d1; ///if the new distance (d1) is smaller than the old distance (d0), update the results ///the length of new path should be closer to t->target_dist if (d1 - mlen/2 < d0 - t->mlen/2) copy = 1; } } if (copy) { t->path_end = out->n-1, t->dist = dist, t->hash = r->hash, t->mlen = mlen, t->is_0 = r->is_0; if (t->target_dist >= 0) { ///src is from li from li to lj, so the dis is generally increased; dijkstra algorithm ///target_dist should be the distance on query if (dist == t->target_dist && t->check_hash && r->hash == t->target_hash) done = 1; else if ((dist > t->target_dist + MG_SHORT_K_EXT) && (dist > (t->target_dist>>4))) done = 1; } } ++t->n_path;///we found a path to the alignment t if (t->n_path >= max_k) done = 1; } if (dst_done->a[off + j] == 0 && done) dst_done->a[off + j] = 1, ++n_done; } ///if all alignments have been settle down ///pre-end; accelerate the loop if (n_done == n_dst) break; } first_src_ban = 1; ///below is used to push new nodes to avl tree for iteration nv = asg_arc_n(g, r->v); av = asg_arc_a(g, r->v); for (i = 0; i < nv; ++i) { // visit all neighbors asg_arc_t *ai = &av[i]; ///v_lv is the (dest_length - overlap_length); it is a normal path length in string graph ///ai->v_lv is the path length from r->v to ai->w ///(r->di>>32) int32_t d = (r->di>>32) + (uint32_t)ai->ul; if (d > max_dist) continue; // don't probe vertices too far away // h keeps visited vertices; path to each visited vertice ///ai->w is the dest ref id; we insert a new ref id, instead of an alignment chain k = kh_put(sp, h, ai->v, &absent);///one node might be visited multiple times q = &kh_val(h, k); if (absent) { // a new vertex visited ///q->k: number of walks from src to ai->w q->k = 0, q->qs = q->qe = -1; q->mlen = 0; ///h_seeds = NULL; so q->mlen = 0 /** //path q->mlen = h_seeds && d + gfa_arc_lw(g, *ai) <= max_dist? node_mlen(km, g, ai->w, &mini, h_seeds, n_seeds, seeds, &q->qs, &q->qe) : 0; **/ //if (ql && qs) fprintf(stderr, "ql=%d,src=%d\tv=%c%s[%d],n_seeds=%d,mlen=%d\n", ql, src, "><"[ai->w&1], g->seg[ai->w>>1].name, ai->w, n_seeds, q->mlen); } ///if there are less than walks from src to ai->w, directly add ///if there are more, keep the smallest walks if (q->k < max_k) { // enough room: add to the heap p = gen_sp_node(km, ai->v, d, id++); p->pre = out->n - 1;///the parent node of this one p->hash = r->hash + __ac_Wang_hash(ai->v); p->is_0 = r->is_0; /** //path if (ai->rank > 0) p->is_0 = 0; **/ kavl_insert(sp, &root, p, 0); q->p[q->k++] = p; ks_heapup_sp(q->k, q->p);///adjust heap by distance } else if ((int32_t)(q->p[0]->di>>32) > d) { // shorter than the longest path so far: replace the longest p = kavl_erase(sp, &root, q->p[0], 0); if (p) { p->di = (uint64_t)d<<32 | (id++); p->pre = out->n - 1; p->hash = r->hash + __ac_Wang_hash(ai->v); p->is_0 = r->is_0; /** //path if (ai->rank > 0) p->is_0 = 0; **/ kavl_insert(sp, &root, p, 0); ks_heapdown_sp(0, q->k, q->p); } else { fprintf(stderr, "Warning: logical bug in gfa_shortest_k(): q->k=%d,q->p[0]->{d,i}={%d,%d},d=%d,src=%u,max_dist=%d,n_dst=%d\n", q->k, (int32_t)(q->p[0]->di>>32), (int32_t)q->p[0]->di, d, src, max_dist, n_dst); km_destroy(km); return; } } // else: the path is longer than all the existing paths ended at ai->w } } kh_destroy(sp, h); // NB: AVL nodes are not deallocated. When km==0, they are memory leaks. for (i = 0, n_found = 0; i < n_dst; ++i) if (dst[i].n_path > 0) ++n_found;///n_path might be larger than 16 ///we can assume n_pathv = NULL for now if (n_found > 0 && res) { // then generate the backtrack array int32_t n; dst_done->n = 0; kv_resize(uint64_t, *dst_done, out->n); uint64_t *trans = dst_done->a; memset(dst_done->a, 0, out->n*sizeof(*(dst_done->a))); // KCALLOC(km, trans, n_out); // used to squeeze unused elements in out[] ///n_out: how many times that nodes in graph have been visited ///note one node might be visited multiples times ///n_dst: number of alignment chains for (i = 0; i < n_dst; ++i) { // mark dst vertices with a target distance mg_path_dst_t *t = &dst[i]; if (t->n_path > 0 && t->target_dist >= 0 && t->path_end >= 0) trans[(uint32_t)out->a[t->path_end]->di] = 1;///(int32_t)out[]->di: traverse track corresponds to the alignment chain dst[] } for (i = 0; (uint32_t)i < out->n; ++i) { // mark dst vertices without a target distance k = kh_get(sp2, h2, out->a[i]->v); if (k != kh_end(h2)) { // TODO: check if this is correct! int32_t off = kh_val(h2, k)>>32, cnt = (int32_t)kh_val(h2, k); for (j = off; j < off + cnt; ++j) if (dst[j].target_dist < 0) trans[i] = 1; } } for (i = (int32_t)(out->n) - 1; i >= 0; --i) // mark all predecessors if (trans[i] && out->a[i]->pre >= 0) trans[out->a[i]->pre] = 1; for (i = n = 0; (uint32_t)i < out->n; ++i) // generate coordinate translations if (trans[i]) trans[i] = n++; else trans[i] = (uint32_t)-1; kv_resize(mg_pathv_t, *res, res->n + n); //res->n += n; for (i = 0; (uint32_t)i < out->n; ++i) { // generate the backtrack array mg_pathv_t *p; if (trans[i] == (uint32_t)-1) continue; p = &res->a[trans[i]+res->n]; p->v = out->a[i]->v, p->d = out->a[i]->di >> 32; p->pre = out->a[i]->pre < 0? out->a[i]->pre:trans[out->a[i]->pre]; } res->n += n; for (i = 0; i < n_dst; ++i) // translate "path_end" if (dst[i].path_end >= 0) dst[i].path_end = trans[dst[i].path_end]; } km_destroy(km); } ///p[]: id of last ///f[]: the score ending at i, not always the peak ///v[]: keeps the peak score up to i; ///t[]: used for buffer ///min_cnt = 2; min_sc = 30; extra_u = 0 ///u = mg_chain_backtrack(n, f, p, v, t, min_cnt, min_sc, 0, &n_u, &n_v); int64_t hc_chain_backtrack(int64_t n, const int64_t *f, const uint64_t *p, uint64_t *srt, uint64_t *u, uint64_t *v, int64_t *n_u_, int64_t *n_v_) { if(n_u_) *n_u_ = 0; if(n_v_) *n_v_ = 0; int64_t i, k, n_v, n_srt, n_v0, n_u, sc; if (n == 0) return 0; // v[] keeps the peak score up to i; f[] is the score ending at i, not always the peak *n_u_ = *n_v_ = 0; for (i = 0, k = 0; i < n; ++i) { if(f[i] >= 0) { srt[k] = (uint64_t)f[i]; srt[k] <<= 32; srt[k] |= ((uint64_t)i)<<1; k++; } } n_srt = k; radix_sort_gfa64(srt, srt + n_srt); ///sort by score ///from the largest to the smallest for (k = n_srt-1, n_v = n_u = 0; k >= 0; --k) { // precompute n_u n_v0 = n_v; for (i = ((uint32_t)srt[k])>>1; i >= 0 && (srt[i]&1) == 0; i = (p[i]==(uint64_t)-1?-1:p[i])) { v[n_v++] = i; srt[i] |= 1; } if(n_v <= n_v0) continue; sc = i < 0? srt[k]>>32: (int64_t)(srt[k]>>32)-f[i]; u[n_u++] = (((uint64_t)sc)<<32) | ((uint64_t)(n_v-n_v0)); } if(n_u_) *n_u_ = n_u; if(n_v_) *n_v_ = n_v; return n_u; } int64_t hc_gchain1_dp(void *km, const ma_ug_t *ug, vec_mg_lchain_t *lc, vec_mg_lchain_t *sw, vec_mg_path_dst_t *dst, vec_sp_node_t *out, vec_mg_pathv_t *path, int64_t qlen, const ug_opt_t *uopt, int64_t bw, double diff_thre, uint64_t *srt, st_mt_t *bf, int64_t *f, uint64_t *p, uint64_t *v) { bf->n = 0; if(lc->n == 0) return 0; int64_t i, j, lc_n = lc->n, n_ext, mm_ovlp, target_dist, max_target_dist, x, m_idx, m_sc; mg_lchain_t *r, *li, *lj; mg_path_dst_t *q; asg_t *g = ug->g; uint64_t isolated, *u; for (i = n_ext = 0; i < lc_n; i++) { r = &lc->a[i]; r->dist_pre = -1; isolated = 0;///dist_pre -> parent in graph chain if((r->re < g->seq[r->v>>1].len) && (r->rs > 0)) isolated = 1;///UL contained in one vertice if (!isolated) ++n_ext; srt[i] = r->qe; srt[i] <<= 32; srt[i] |= (uint64_t)i; srt[i] |= (isolated<<63); } radix_sort_gfa64(srt, srt+lc_n); for (i = 1, j = 0; i <= lc_n; i++) { if (i == lc_n || (srt[i]>>32) != (srt[j]>>32)) { if(i - j > 1) { for (x = j; x < i; x++) { srt[x] <<= 32; srt[x] >>= 32; srt[x] |= ((uint64_t)lc->a[(uint32_t)srt[x]].qs)<<32; } radix_sort_gfa64(srt+j, srt+i); } j = i; } } kv_resize(mg_lchain_t, *sw, (uint64_t)lc_n); sw->n = lc_n; for (i = 0; i < lc_n; i++) sw->a[i] = lc->a[(uint32_t)srt[i]]; memcpy(lc->a, sw->a, lc_n *sizeof((*(lc->a)))); // fprintf(stderr, "[M::%s::] n_ext:%ld, lc_n:%ld\n", __func__, n_ext, lc_n); for (i = 0; i < n_ext; ++i) { // core loop li = &lc->a[i]; mm_ovlp = max_ovlp(g, li->v^1); x = (li->qs + mm_ovlp)*diff_thre; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > qlen+1) x = qlen+1; x = find_mg_lchain_max(i, lc->a, x+G_CHAIN_INDEL); // fprintf(stderr, "\nli->(%ld)\tutg%.6d%c(%u)\tqs:%u\tqe:%u\t%c\trs:%u\tre:%u\tsrc:%u\tscore:%d, x:%ld\n", // i, (int32_t)(li->v>>1)+1, "lc"[ug->u.a[li->v>>1].circ], ug->u.a[li->v>>1].len, // li->qs, li->qe, "+-"[li->v&1], li->rs, li->re, li->v^1, li->score, x); // collect potential destination vertices for (dst->n = 0, max_target_dist= -1, j = x; j >= 0; --j) { lj = &lc->a[j]; ///extend_end_coord(lj, qlen, g->seq[lj->v>>1].len, &jqs, &jqe, &jrs, &jre); //lj contained in li; actually in circle, this might happen; need to deal with it later if(lj->qs >= li->qs+G_CHAIN_INDEL) continue; ///if there is a circle, the two linear chains might be at the same vertice target_dist = hc_target_len(g, li, lj); // fprintf(stderr, "j:%ld, target_dist:%ld\n", j, target_dist); if(target_dist < 0) continue; kv_pushp(mg_path_dst_t, *dst, &q); memset(q, 0, sizeof(*q)); q->inner = 0;//we set q->inner = 0 to allow circles q->v = lj->v^1;///must be v^1 instead of v q->meta = j; ///lj->qs************lj->qe /// li->qs************li->qe q->qlen = li->qs - lj->qe;///might be negative; this is the region that need to be checked in base-level q->target_dist = target_dist;///cannot understand the target_dist q->target_hash = 0; q->check_hash = 0; if(max_target_dist < target_dist) max_target_dist = target_dist; ///not sure how to use this cut-off // if (t[j] == i) { // if (++n_skip > max_skip) // break; // } // if (p[j] >= 0) t[p[j]] = i; // if((li->v>>1)==10 && ((lj->v>>1)==15||(lj->v>>1)==14)) max_target_dist = 100000; // fprintf(stderr, "+++lj->(%ld)\tutg%.6d%c(%u)\t%u\t%u\t%c\ttarget_dist:%d\n", // j, (int32_t)(lj->v>>1)+1, "lc"[ug->u.a[lj->v>>1].circ], ug->u.a[lj->v>>1].len, // lj->qs, lj->qe, "+-"[lj->v&1], q->target_dist); } // confirm reach-ability int64_t max_f = li->score, max_j = -1, max_d = -1, max_inner = 0; uint32_t max_hash = 0; if(dst->n) { max_target_dist *= (1+diff_thre); if(max_target_dist < bw) max_target_dist = bw; hc_shortest_k(km, g, li->v^1, dst->n, dst->a, max_target_dist, MG_MAX_SHORT_K, bf, srt, out, NULL, 1); // remove unreachable destinations //TODO: check sequence identity for (j = 0; j < (int64_t)dst->n; ++j) { mg_path_dst_t *dj = &dst->a[j]; int32_t sc; if (dj->n_path == 0) continue; // unreachable sc = cal_gchain_sc(dj, li, lc->a, f, bw, diff_thre, W_CHN_PEN_GAP); // fprintf(stderr, "---dj->(%ld)\tutg%.6d%c(%u)\tsc:%d\tmax_f:%ld\ttarget_dist:%d\tdj->dist:%d\n", // j, (int32_t)(dj->v>>1)+1, "lc"[ug->u.a[dj->v>>1].circ], ug->u.a[dj->v>>1].len, sc, max_f, dj->target_dist, dj->dist); if (sc == INT32_MIN) continue; // out of band // fprintf(stderr, "+max_f->%d, max_j->%d\n", max_f, max_j); if (sc < 0) continue;// negative score // fprintf(stderr, "++max_f->%d, max_j->%d\n", max_f, max_j); if (sc > max_f) { max_f = sc, max_j = dj->meta, max_d = dj->dist, max_hash = dj->hash, max_inner = dj->inner; // fprintf(stderr, "+++max_f->%d, max_j->%d\n", max_f, max_j); } } } f[i] = max_f, p[i] = max_j<0?(uint64_t)-1:max_j; li->dist_pre = max_d; li->hash_pre = max_hash; li->inner_pre = max_inner; // fprintf(stderr, "i->%ld, utg%.6d%c->utg%.6d%c, max_f:%ld\n", i, (int32_t)(li->v>>1)+1, "lc"[ug->u.a[li->v>>1].circ], // max_j<0?0:(int32_t)(lc->a[max_j].v>>1)+1, max_j<0?'*':"lc"[ug->u.a[lc->a[max_j].v>>1].circ], max_f); } int64_t k, k0, n_u, n_v, ni; kv_resize(uint64_t, *bf, (uint64_t)lc_n); u = bf->a; hc_chain_backtrack(n_ext, f, p, srt, u, v, &n_u, &n_v); for (i = 0; i < lc_n - n_ext; ++i) { u[n_u++] = (((uint64_t)lc->a[n_ext + i].score)<<32) | 1; v[n_v++] = n_ext + i; } sw->n = 0; kv_resize(mg_lchain_t, *sw, (uint64_t)n_v); m_idx = m_sc = -1; for (i = 0, k = 0; i < n_u; ++i) { k0 = k, ni = (int32_t)u[i]; for (j = 0; j < ni; ++j) { sw->a[k++] = lc->a[v[k0 + (ni - j - 1)]]; } if(m_idx < 0 || m_sc < ((int64_t)(u[i]>>32))) { m_idx = i; m_sc = ((int64_t)(u[i]>>32)); } } assert(k == n_v); bf->n = n_u; memcpy(lc->a, sw->a, n_v*sizeof(mg_lchain_t)); return m_idx; } void debug_gchain(const asg_t *g, mg_lchain_t *a, uint64_t n) { uint64_t k, i, v, w, nv; asg_arc_t *av; for (k = 1; k < n; k++) { v = a[k-1].v; w = a[k].v; nv = asg_arc_n(g, v); av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].v == w) break; } if(i >= nv) { // fprintf(stderr, "[M::%s::]\n", __func__); fprintf(stderr, "[M::%s::]\tutg%.6dl(%c)\t->\tutg%.6dl(%c)\n", __func__, (int32_t)(v>>1)+1, "+-"[v&1], (int32_t)(w>>1)+1, "+-"[w&1]); } } } void debug_gchain2(const asg_t *g, mg_pathv_t *a, uint64_t n) { uint64_t k, i, v, w, nv; asg_arc_t *av; for (k = 1; k < n; k++) { v = a[k-1].v; w = a[k].v; nv = asg_arc_n(g, v); av = asg_arc_a(g, v); for (i = 0; i < nv; i++) { if(av[i].v == w) break; } if(i >= nv) { // fprintf(stderr, "[M::%s::]\n", __func__); fprintf(stderr, "[M::%s::]\tutg%.6dl(%c)\t->\tutg%.6dl(%c)\n", __func__, (int32_t)(v>>1)+1, "+-"[v&1], (int32_t)(w>>1)+1, "+-"[w&1]); } } } void reverse_track(mg_pathv_t *a, uint64_t a_n) { uint64_t k; mg_pathv_t z; for (k = 0; k < (a_n>>1); k++) { z = a[k]; a[k] = a[a_n - k - 1]; a[a_n - k - 1] = z; a[k].v ^= 1; a[a_n - k - 1].v ^= 1; } if(a_n&1) a[k].v ^= 1; } uint32_t gen_gchain_track(void *km, mg_lchain_t *a, int64_t a_n, const asg_t *g, st_mt_t *dst_done, vec_sp_node_t *out, vec_mg_pathv_t *res) { int64_t k, p_n; mg_lchain_t *l0, *l1; mg_path_dst_t dst; uint64_t dst_group; mg_pathv_t *p; res->n = 0; kv_pushp(mg_pathv_t, *res, &p); p->v = p->d = (uint32_t)-1; p->d = 0; for (k = 1; k < a_n; k++) { l0 = a + k - 1; l1 = a + k; assert(!l1->inner_pre); memset(&dst, 0, sizeof(dst)); dst.v = l0->v^1; assert(l1->dist_pre >= 0); dst.target_dist = l1->dist_pre; dst.target_hash = l1->hash_pre; dst.check_hash = 1; p_n = res->n; hc_shortest_k(km, g, l1->v^1, 1, &dst, dst.target_dist, MG_MAX_SHORT_K, dst_done, &dst_group, out, res, 1); // debug_gchain2(g, res->a + p_n, res->n - p_n); // fprintf(stderr, "[M::%s::n->%ld]\tutg%.6dl(%c)\t->\tutg%.6dl(%c)\n", __func__, res->n - p_n, // (int32_t)(l0->v>>1)+1, "+-"[l0->v&1], (int32_t)(l1->v>>1)+1, "+-"[l1->v&1]); assert(res->n - p_n > 1); assert(dst.target_hash == dst.hash); res->n--; reverse_track(res->a + p_n, res->n - p_n); res->n--;///reomve l1 from res kv_pushp(mg_pathv_t, *res, &p); p->v = p->d = (uint32_t)-1; p->d = k; } return res->n; } void print_chain(mg_lchain_t *a, uint32_t a_n) { uint32_t k; for (k = 0; k < a_n; k++) { if(a[k].off!=-1) { fprintf(stderr, "%u\t%u\t%c\tutg%.6dl\t%u\t%u\n", a[k].qs, a[k].qe, "+-"[a[k].v&1], (int32_t)(a[k].v>>1)+1, a[k].rs, a[k].re); } else { fprintf(stderr, "*\t*\t%c\tutg%.6dl\t*\t*\n", "+-"[a[k].v&1], (int32_t)(a[k].v>>1)+1); } } } void dedup_second_chain(uint64_t *a, int64_t a_n, int64_t p_sidx, int64_t p_eidx, mg_lchain_t *chain_a, kv_ul_ov_t *raw_idx, kv_ul_ov_t *raw_chn) { int64_t k, i; for (k = p_sidx; k < p_eidx; k++) { i = raw_idx->a[chain_a[k].off].tn; for (;i>=0;) { i = raw_chn->a[i].qn == (uint32_t)-1?-1:raw_chn->a[i].qn; } } } uint32_t gen_max_gchain(void *km, int64_t ulid, st_mt_t *idx, vec_mg_lchain_t *e, int64_t qlen, float primary_cov_rate, float primary_fragment_cov_rate, float primary_fragment_second_score_rate, const asg_t *g, st_mt_t *dst_done, vec_sp_node_t *out, vec_mg_pathv_t *res) { if(idx->n <= 0) return 0; int64_t a_n, idx_n = idx->n, i, m_sc = 0, is_done = 0; uint64_t s_idx, e_idx, om, ok, ovlp; ul_ov_t m; memset(&m, 0, sizeof(m)); m_sc = -1; mg_lchain_t *a = e->a; for (i = a_n = 0; i < idx_n; ++i) { if(((int64_t)(idx->a[i]>>32)) > m_sc) { m_sc = ((int64_t)(idx->a[i]>>32)); m.qn = i; m.ts = a_n; m.te = a_n + ((uint32_t)idx->a[i]); m.qs = a[m.ts].qs; m.qe = a[m.te-1].qe; } a_n += ((uint32_t)idx->a[i]); } assert(a[m.ts].qs<=a[m.te-1].qs && a[m.te-1].qe>=a[m.ts].qe); // print_chain(a + m.ts, m.te - m.ts); if((m.qe - m.qs) > (qlen*primary_cov_rate)) is_done = 1; if(is_done == 0) { for (i = a_n = 0; i < idx_n; ++i) { s_idx = a[a_n].qs; a_n += ((uint32_t)idx->a[i]); e_idx = a[a_n-1].qe; if(i == m.qn) continue; if(s_idx < m.qs || e_idx < m.qs || s_idx > m.qe || e_idx > m.qe) break; } if(i >= idx_n) is_done = 2;///no alignment that is on the left or the right side of the primary chain } if(is_done == 0) { if((m.qe - m.qs) > (qlen*primary_fragment_cov_rate)) { om = m.qe - m.qs; for (i = a_n = 0; i < idx_n; ++i) { s_idx = a[a_n].qs; a_n += ((uint32_t)idx->a[i]); e_idx = a[a_n-1].qe; if(i == m.qn) continue; ovlp = ((MIN(m.qe, e_idx) > MAX(m.qs, s_idx))? (MIN(m.qe, e_idx) - MAX(m.qs, s_idx)):0); if(ovlp == 0) continue; ok = e_idx - s_idx; if(ok > om ) ok = om; if((ovlp > ok*0.25) && ((int64_t)(idx->a[i]>>32)) > (m_sc*primary_fragment_second_score_rate)) break; } if(i >= idx_n) is_done = 3; } } if(is_done && gen_gchain_track(km, a + m.ts, m.te - m.ts, g, dst_done, out, res)) {///try to find a path for (i = m.ts, e->n = 0; i < (int64_t)m.te; i++) a[e->n++] = a[i]; // fprintf(stderr, "--[M::%s::id->%ld] [%u, %u), res->n:%lu\n", __func__, ulid, m.qs, m.qe, (uint64_t)res->n); kv_resize(mg_lchain_t, *e, res->n); a = e->a; for (i = ((int64_t)res->n)-1; i >= 0; i--) { if(res->a[i].v == (uint32_t)-1) { a[i] = a[res->a[i].d]; // fprintf(stderr, "ulid:%ld\t%u\t%u\t%c\tutg%.6dl\t%u\t%u\n", ulid, a[i].qs, a[i].qe, "+-"[a[i].v&1], (int32_t)(a[i].v>>1)+1, a[i].rs, a[i].re); } else { a[i].v = res->a[i].v; a[i].off = -1; // fprintf(stderr, "ulid:%ld\t*\t*\t%c\tutg%.6dl\t*\t*\n", ulid, "+-"[a[i].v&1], (int32_t)(a[i].v>>1)+1); } } e->n = res->n; // debug_gchain(g, e->a, e->n); return 1; } return 0; } void update_ul_vec_t(ul_vec_t *rch, vec_mg_lchain_t *u, const ul_idx_t *uref) { uint64_t k; ma_ug_t *ug = uref->ug; for (k = 0; k < u->n; k++) { if(u->a[k].off != -1) { fprintf(stderr, "(%lu) %u\t%u\t%c\tutg%.6d%c\t%u\t%u\n", k, u->a[k].qs, u->a[k].qe, "+-"[u->a[k].v&1], (int32_t)(u->a[k].v>>1)+1, "lc"[ug->u.a[u->a[k].v>>1].circ], u->a[k].rs, u->a[k].re); } else { fprintf(stderr, "(%lu) *\t*\t%c\tutg%.6d%c\t*\t*\n", k, "+-"[u->a[k].v&1], (int32_t)(u->a[k].v>>1)+1, "lc"[ug->u.a[u->a[k].v>>1].circ]); } } } ///sps and hap are just vector for uint64_t; used for buffer uint32_t direct_gchain(mg_tbuf_t *b, ul_vec_t *rch, glchain_t *ll, gdpchain_t *gdp, st_mt_t *sps, haplotype_evdience_alloc *hap, const ul_idx_t *uref, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, int64_t max_skip, int64_t ulid) { // if(ulid!=108) return 0; kv_ul_ov_t *idx = &(ll->lo), *init = &(ll->tk); int64_t max_idx; idx->n = init->n = 0; gl_rg2ug_gen(rch, idx, uref, 1); if(idx->n == 0) return 0; ///generate linear chains gen_linear_chains(idx, init, uref, uopt, bw, diff_ec_ul, rch->rlen, max_skip, ll, sps); assert(idx->n); if(idx->n == 0) return 0; fprintf(stderr, "\n++[M::%s::%.*s(id:%ld), len:%u] idx->n:%lu\n", __func__, UL_INF.nid.a[ulid].n, UL_INF.nid.a[ulid].a, ulid, rch->rlen, (uint64_t)idx->n); dump_linear_chain(uref->ug->g, idx, init, &(gdp->l), rch->rlen); // fprintf(stderr, "\n+++[M::%s::id->%ld, len->%u] idx->n:%lu\n", __func__, ulid, rch->rlen, (uint64_t)idx->n); // kv_resize(uint64_t, ll->srt.a, idx->n); kv_resize(uint64_t, hap->snp_srt, idx->n); kv_resize(uint64_t, gdp->v, idx->n); // occ = gl_chain_advance(&(gdp->l), &(gdp->swap), uref, uopt, G_CHAIN_BW, diff_ec_ul, qlen, UG_SKIP, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, G_CHAIN_TRANS_WEIGHT, 0, NULL, uref->ug, debug_i, km); ///buffer kv_resize(uint64_t, ll->srt.a, gdp->l.n); kv_resize(uint64_t, hap->snp_srt, gdp->l.n); kv_resize(uint64_t, gdp->v, gdp->l.n); kv_resize(int64_t, gdp->f, gdp->l.n); max_idx = hc_gchain1_dp(b->km, uref->ug, &(gdp->l), &(gdp->swap), &(gdp->dst), &(gdp->out), &(gdp->path), rch->rlen, uopt, bw, diff_ec_ul, ll->srt.a.a, sps, gdp->f.a, hap->snp_srt.a, gdp->v.a); // fprintf(stderr, "++++[M::%s::id->%ld, len->%u] gdp->l.n:%lu\n", __func__, ulid, rch->rlen, (uint64_t)gdp->l.n); // fprintf(stderr, "+[M::%s::] gdp->l.n:%lu\n", __func__, (uint64_t)gdp->l.n); //sps has the chain idx; gdp->l has the chain if(max_idx >= 0 && gen_max_gchain(b->km, ulid, sps, &(gdp->l), rch->rlen, P_CHAIN_COV, P_FRAGEMENT_PRIMARY_CHAIN_COV, P_FRAGEMENT_PRIMARY_SECOND_COV, uref->ug->g, &(gdp->dst_done), &(gdp->out), &(gdp->path))) { // update_ul_vec_t(rch, &(gdp->l), uref); // __ac_X31_hash_string("hehe"); } else { uint64_t i; fprintf(stderr, "unsuccess->[M::%s::id->%ld, len->%u] gdp->l.n:%lu\n", __func__, ulid, rch->rlen, (uint64_t)gdp->l.n); for (i = 0; i < gdp->l.n; ++i) { fprintf(stderr, "(%lu)\tutg%.6d%c(%u)\t%u\t%u\t%c\tsrc:%u\tscore:%d\n", i, (int32_t)(gdp->l.a[i].v>>1)+1, "lc"[uref->ug->u.a[gdp->l.a[i].v>>1].circ], uref->ug->u.a[gdp->l.a[i].v>>1].len, gdp->l.a[i].qs, gdp->l.a[i].qe, "+-"[gdp->l.a[i].v&1], gdp->l.a[i].v^1, gdp->l.a[i].score); } } // occ = gl_chain_advance(idx, ll->tk.a+ll->tk.n, uref, uopt, G_CHAIN_BW, diff_ec_ul, qlen, UG_SKIP, dumy->overlapID, ll->srt.a.a, hap->snp_srt.a, G_CHAIN_TRANS_WEIGHT, 0, NULL, uref->ug, debug_i, km); // simple_g_chain_dp(idx, buf->a, uref, uopt, bw, diff_ec_ul, rch->rlen, max_skip, ll->srt.a.a, hap->snp_srt.a, sps->a); // if(check_extension_end(idx, rch->rlen, buf->a)) { // // ug2rg_gen(idx->a[idx->n-1].qs, idx->a[idx->n-1].qe, buf->a + idx->a[idx->n-1].ts, idx->a[idx->n-1].te - idx->a[idx->n-1].ts, rch); // } else {///need graph chaining // } return 0; } static void worker_for_ul_gchains_alignment(void *data, long i, int tid) { ul_vec_t *p = &(UL_INF.a[i]); if(p->dd == 1) return; //fully aligned if(p->bb.n == 1 && p->bb.a[0].base) return;///no alignment utepdat_t *s = (utepdat_t*)data; direct_gchain(s->buf[tid], p, &(s->ll[tid]), &(s->gdp[tid]), &(s->sps[tid]), &(s->hab[tid]->hap), s->uu, s->uopt, G_CHAIN_BW, s->opt->diff_ec_ul, UG_SKIP, i); // gl_chain_refine_advance(&b->olist, &b->correct, &b->hap, bl, s->uu, s->opt->diff_ec_ul, winLen, s->len[i], s->uopt, s->id+i, km); } void work_ul_gchains(uldat_t *sl) { utepdat_t s; uint64_t i; memset(&s, 0, sizeof(s)); s.id = 0; s.opt = sl->opt; s.ug = sl->ug; s.uopt = sl->uopt; s.rg = sl->rg; s.uu = sl->uu; CALLOC(s.hab, sl->n_thread); CALLOC(s.buf, sl->n_thread); CALLOC(s.ll, sl->n_thread); CALLOC(s.gdp, sl->n_thread); CALLOC(s.mzs, sl->n_thread); CALLOC(s.sps, sl->n_thread); for (i = 0; i < sl->n_thread; ++i) { s.hab[i] = ha_ovec_init(0, 0, 1); s.buf[i] = mg_tbuf_init(); } kt_for(sl->n_thread, worker_for_ul_gchains_alignment, &s, UL_INF.n); for (i = 0; i < sl->n_thread; ++i) { ha_ovec_destroy(s.hab[i]); mg_tbuf_destroy(s.buf[i]); hc_glchain_destroy(&(s.ll[i])); hc_gdpchain_destroy(&(s.gdp[i])); kv_destroy(s.mzs[i]); kv_destroy(s.sps[i]); } free(s.hab); free(s.buf); free(s.ll); free(s.gdp); free(s.mzs); free(s.sps); } void print_ul_ovlps(all_ul_t *x, int32_t prt_ovlp) { uint64_t k, i, ucov_occ = 0, cov_occ = 0, ucov_len = 0, cov_len = 0, unaligned_len = 0, unaligned_occ = 0, aligned_occ = 0; ul_vec_t *p = NULL; nid_t *z = NULL; uc_block_t *m = NULL; for (k = 0; k < x->n; k++) { z = &(x->nid.a[k]); p = &(x->a[k]); fprintf(stderr, "S\t%.*s\tq:id:%lu\tl:%u\tdd:%d\n", (int32_t)z->n, z->a, k, p->rlen, p->bb.n == 1&&p->bb.a[0].base?-1:(int32_t)p->dd); if(prt_ovlp) { for (i = 0; i < p->bb.n; i++) { m = &(p->bb.a[i]); if(m->base) { ucov_occ++; ucov_len += (m->qe-(m->hid&FLANK_M)) - (m->qs+((m->hid>>15)&FLANK_M)); fprintf(stderr, "B\t%.*s\t%u\t%u\t%u\n", (int32_t)z->n, z->a, p->rlen, (m->qs+((m->hid>>15)&FLANK_M)), (m->qe-(m->hid&FLANK_M))); } else { fprintf(stderr, "A\t%.*s\t%u\t%u\t%u\t%c\t%.*s\t%u\t%u\t%u\n", (int32_t)z->n, z->a, p->rlen, m->qs, m->qe, "+-"[m->rev], (int32_t)Get_NAME_LENGTH(R_INF, m->hid), Get_NAME(R_INF, m->hid), (uint32_t)Get_READ_LENGTH(R_INF, m->hid), m->ts, m->te); if(m->el) cov_occ++; } } } if(p->bb.n == 1 && p->bb.a[0].base) { unaligned_len += p->rlen; unaligned_occ++; } else { aligned_occ++; } cov_len += p->rlen; } cov_len -= ucov_len; fprintf(stderr, "[M::%s::] ==>aligned_occ:%lu, unaligned_occ:%lu\n", __func__, aligned_occ, unaligned_occ); fprintf(stderr, "[M::%s::] ==>cov_len:%lu, ucov_len:%lu, unaligned_len:%lu\n", __func__, cov_len, ucov_len-unaligned_len, unaligned_len); } void print_all_ul_t_stat(all_ul_t *x) { uint64_t k, i, ucov_occ = 0, cov_occ = 0, ucov_len = 0, cov_len = 0; ul_vec_t *p = NULL; for (k = 0; k < x->n; k++) { p = &(x->a[k]); for (i = 0; i < p->bb.n; i++) { if(p->bb.a[i].base/**.hid&x->mm**/) { ucov_occ++; ucov_len += (p->bb.a[i].qe-(p->bb.a[i].hid&FLANK_M)) - (p->bb.a[i].qs+((p->bb.a[i].hid>>15)&FLANK_M)); } else { cov_occ++; } } cov_len += p->rlen; } cov_len -= ucov_len; fprintf(stderr, "[M::%s::] ==>cov_occ:%lu, ucov_occ:%lu\n", __func__, cov_occ, ucov_occ); fprintf(stderr, "[M::%s::] ==>cov_len:%lu, ucov_len:%lu\n", __func__, cov_len, ucov_len); } void print_ovlp_src_bl_stat(all_ul_t *x, const ug_opt_t *uopt) { uint64_t k, z, tc, ta; ma_hit_t_alloc* src = uopt->sources; for (k = tc = ta = 0; k < R_INF.total_reads; k++) { if(x->ridx.idx.a[k+1] - x->ridx.idx.a[k] == 0) continue; tc++; for (z = 0; z < src[k].length; z++) { if(src[k].buffer[z].bl) { ta++; break; } } } fprintf(stderr, "[M::%s::] ==> # HiFi reads:%lu, # covered HiFi reads:%lu, # chained HiFi reads:%lu\n", __func__, R_INF.total_reads, tc, ta); uint64_t tt[4] = {0}; for (k = 0; k < x->n; k++) tt[x->a[k].dd]++; fprintf(stderr, "[M::%s::] ==> # passed UL reads:%lu, # fully corrected UL reads:%lu, # almost fully corrected UL reads:%lu, # UL reads have primary chains:%lu\n", __func__, tt[0]+tt[1]+tt[2]+tt[3], tt[1], tt[2], tt[3]); } void gen_ul_vec_rid_t(all_ul_t *x) { ul_vec_rid_t *ridx = &(x->ridx); uint64_t k, i, l, m, *a, a_n; ul_vec_t *p = NULL; ridx->idx.n = ridx->idx.m = R_INF.total_reads + 1; CALLOC(ridx->idx.a, ridx->idx.n); for (k = 0; k < x->n; k++) { p = &(x->a[k]); for (i = 0; i < p->bb.n; i++) { if(p->bb.a[i].base) continue; ridx->idx.a[p->bb.a[i].hid]++; } } for (k = l = 0; k < ridx->idx.n; k++) { m = ridx->idx.a[k]; ridx->idx.a[k] = l; l += m; } ridx->occ.n = ridx->occ.m = l; MALLOC(ridx->occ.a, ridx->occ.n); for (k = 0; k < R_INF.total_reads; k++) { a = ridx->occ.a + ridx->idx.a[k]; a_n = ridx->idx.a[k+1] - ridx->idx.a[k]; if(a_n) a[a_n-1] = 0; } for (k = 0; k < x->n; k++) { p = &(x->a[k]); for (i = 0; i < p->bb.n; i++) { if(p->bb.a[i].base) continue; a = ridx->occ.a + ridx->idx.a[p->bb.a[i].hid]; a_n = ridx->idx.a[p->bb.a[i].hid+1] - ridx->idx.a[p->bb.a[i].hid]; if(a_n) { if(a[a_n-1] == a_n-1) a[a_n-1] = (k<<32)|i; else a[a[a_n-1]++] = (k<<32)|i; } } } } int32_t find_ul_block_max_reverse(int32_t n, const uc_block_t *a, uint32_t x) { int32_t s = 0, e = n; if (n == 0) return n; if (a[0].qe < x) return 0;///max qe if (a[n-1].qe >= x) return n;///min qe while (e > s) { // TODO: finish this block int32_t m = s + (e - s) / 2; // if (a[m].qe >= x) e = m; // else s = m + 1; if (a[m].qe > x) s = m + 1; else e = m; } assert(s == e); return s; } int32_t find_ul_block_max(int32_t n, const uc_block_t *a, uint32_t x) { int32_t s = 0, e = n; if (n == 0) return -1; if (a[n-1].qe < x) return n - 1; if (a[0].qe >= x) return -1; while (e > s) { // TODO: finish this block int32_t m = s + (e - s) / 2; if (a[m].qe >= x) e = m; else s = m + 1; } assert(s == e); return s; } /** void determine_connective(all_ul_t *m, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, ul_vec_t *p, uint32_t ii, uint64_t rid) { if((p->bb.a[ii].base) || p->bb.a[ii].hid != rid) fprintf(stderr, "ERROR\n"); if(p->bb.n <= ii + 1) return; uint32_t li_v, lk_v, k, ol; int64_t mm_ovlp, x; uc_block_t *li = NULL, *lk = NULL; ma_hit_t *t = NULL; li = &(p->bb.a[ii]); li_v = (((uint32_t)(li->hid))<<1)|((uint32_t)(li->rev)); mm_ovlp = max_ovlp_src(uopt, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > p->rlen+1) x = p->rlen+1; x = find_ul_block_max_rev(p->bb.n - ii - 1, p->bb.a + ii + 1, x) + ii + 1; for (k = x; k < p->bb.n; ++k) { // collect potential destination vertices lk = &(p->bb.a[k]); lk_v = (((uint32_t)(lk->hid))<<1)|((uint32_t)(lk->rev)); if(lk->qe <= li->qs) break;//even this pair has a overlap, its length will be very small; just ignore if((li_v == lk_v) || (lk->base)) continue; // if(li->qs <= 0) continue;///means the UL read does not longer than the overlap between li and lk // if(lk->qs <= 0) continue;//the UL read should be cover the whole HiFi reads li and lk if(((li->te - li->ts)*1.05) < Get_READ_LENGTH(R_INF, li->hid)) continue; if(((lk->te - lk->ts)*1.05) < Get_READ_LENGTH(R_INF, lk->hid)) continue; x = infer_rovlp(NULL, NULL, li, lk, &R_INF, NULL); t = query_ovlp_src(uopt, li_v^1, lk_v^1, x, diff_ec_ul, &ol); if(t) { // sum = t->bl + ol; // t->bl = (sum & 0x7fffffffU); t->bl++; } } } **/ ///note: we only label reliable chains void determine_connective_adv(all_ul_t *m, const ug_opt_t *uopt, int64_t bw, double diff_ec_ul, ul_vec_t *p, uint32_t ii, uint64_t rid) { assert((!p->bb.a[ii].base)&&(p->bb.a[ii].hid == rid)); if(ii <= 0) return; if(!(p->bb.a[ii].pchain)) return; ///not a primary chain if(!(p->bb.a[ii].el)) return; ///not a cis alignment uint32_t li_v, lk_v, ol; int64_t mm_ovlp, k, x; uc_block_t *li = NULL, *lk = NULL; ma_hit_t *t = NULL; li = &(p->bb.a[ii]); li_v = (((uint32_t)(li->hid))<<1)|((uint32_t)(li->rev)); mm_ovlp = max_ovlp_src(uopt, li_v^1); x = (li->qs + mm_ovlp)*diff_ec_ul; if(x < bw) x = bw; x += li->qs + mm_ovlp; if (x > p->rlen+1) x = p->rlen+1; x = find_ul_block_max(ii, p->bb.a, x+G_CHAIN_INDEL); for (k = x; k >= 0; --k) { // collect potential destination vertices lk = &(p->bb.a[k]); lk_v = (((uint32_t)(lk->hid))<<1)|((uint32_t)(lk->rev)); if(lk->qe+G_CHAIN_INDEL <= li->qs) break;//even this pair has a overlap, its length will be very small; just ignore if(lk->base || (!(lk->pchain)) || (!(lk->el))) continue; if(li_v == lk_v) continue; // if(li->qs <= 0) continue;///means the UL read does not longer than the overlap between li and lk // if(lk->qs <= 0) continue;//the UL read should be cover the whole HiFi reads li and lk // if(((li->te - li->ts)*1.05) < Get_READ_LENGTH(R_INF, li->hid)) continue; // if(((lk->te - lk->ts)*1.05) < Get_READ_LENGTH(R_INF, lk->hid)) continue; if((li->te - li->ts) < Get_READ_LENGTH(R_INF, li->hid)) continue; if((lk->te - lk->ts) < Get_READ_LENGTH(R_INF, lk->hid)) continue; x = /**((int64_t)(lk->qe))-((int64_t)(li->qs))**/infer_rovlp(NULL, NULL, li, lk, &R_INF, NULL); t = query_ovlp_src(uopt, li_v^1, lk_v^1, x, diff_ec_ul, &ol); if(t) { // sum = t->bl + ol; // t->bl = (sum & 0x7fffffffU); t->bl++; } } } static void update_ovlp_src(void *data, long i, int tid) // callback for kt_for() { uldat_t *sl = (uldat_t *)data; ma_hit_t_alloc* src = sl->uopt->sources; uint64_t z, k, *a, a_n; for (z = 0; z < src[i].length; z++) src[i].buffer[z].bl = 0; a = UL_INF.ridx.occ.a + UL_INF.ridx.idx.a[i]; a_n = UL_INF.ridx.idx.a[i+1] - UL_INF.ridx.idx.a[i]; for (k = 0; k < a_n; k++) { ///note: we only label reliable chains determine_connective_adv(&UL_INF, sl->uopt, G_CHAIN_BW, sl->opt->diff_ec_ul, &(UL_INF.a[a[k]>>32]), (uint32_t)(a[k]), i); // determine_connective(&UL_INF, sl->uopt, G_CHAIN_BW, sl->opt->diff_ec_ul, // &(UL_INF.a[a[k]>>32]), (uint32_t)(a[k]), i); } } uint64_t* get_hifi2ul_list(all_ul_t *x, uint64_t hid, uint64_t* a_n) { (*a_n) = x->ridx.idx.a[hid+1] - x->ridx.idx.a[hid]; return x->ridx.occ.a + x->ridx.idx.a[hid]; } static void update_ovlp_src_bl(void *data, long i, int tid) { uldat_t *sl = (uldat_t *)data; ma_hit_t_alloc* src = sl->uopt->sources; uint64_t z, sum; uint32_t qn, tn; int32_t idx; for (z = 0; z < src[i].length; z++) { qn = Get_qn(src[i].buffer[z]); tn = Get_tn(src[i].buffer[z]); if(qn > tn) continue; idx = get_specific_overlap(&(src[tn]), tn, qn); assert(idx != -1); sum = src[i].buffer[z].bl + src[tn].buffer[idx].bl; src[i].buffer[z].bl = src[tn].buffer[idx].bl = sum/**(sum&0x7fffffffU)**/; } } int scall_ul_pipeline(uldat_t* sl, const enzyme *fn) { double index_time = yak_realtime(); int i; init_all_ul_t(&UL_INF, &R_INF); for (i = 0; i < fn->n; i++){ gzFile fp; if ((fp = gzopen(fn->a[i], "r")) == 0) return 0; sl->ks = kseq_init(fp); kt_pipeline(3, worker_ul_scall_pipeline, sl, 3); kseq_destroy(sl->ks); gzclose(fp); } sl->hits.total_base = sl->total_base; sl->hits.total_pair = sl->total_pair; fprintf(stderr, "[M::%s::%.3f] ==> Qualification\n", __func__, yak_realtime()-index_time); fprintf(stderr, "[M::%s::] ==> # reads: %lu, # bases: %lu\n", __func__, UL_INF.n, sl->total_base); fprintf(stderr, "[M::%s::] ==> # bases: %lu; # corrected bases: %lu; # recorrected bases: %lu\n", __func__, sl->num_bases, sl->num_corrected_bases, sl->num_recorrected_bases); gen_ul_vec_rid_t(&UL_INF); return 1; } int print_ul_rs(all_ul_t *U_INF) { uint32_t i; UC_Read ur; init_UC_Read(&ur); for (i = 0; i < U_INF->n; i++) { retrieve_ul_t(&ur, NULL, U_INF, i, 0, 0, -1); fprintf(stderr, ">%s\n", U_INF->nid.a[i].a); fprintf(stderr, "%.*s\n", (int)ur.length, ur.seq); } destory_UC_Read(&ur); return 1; } inline void get_ulname(mg_dbn_t *name, int32_t rid, char **rn, int32_t *rl) { (*rn) = name->cc.a + (rid>0?name->a[rid-1]:0); (*rl) = name->a[rid] - (rid>0?name->a[rid-1]:0); } void print_gaf(const ma_ug_t *ug, mg_gres_a *hits, mg_dbn_t *name) { uint64_t i, q; int32_t k, nl, m; char *nn; mg_gchain_t *gc; mg_lres_t *lc; for (i = 0; i < hits->n; i++) { q = hits->a[i].qid; nn = name->cc.a + (q>0?name->a[q-1]:0); nl = name->a[q] - (q>0?name->a[q-1]:0); for (k = 0; k < hits->a[i].n_gc; k++) { gc = &(hits->a[i].gc[k]); fprintf(stderr, "S\t%.*s\tq:id:%lu\tl:n:%d\n", nl, nn, q, gc->cnt); for (m = 0; m < gc->cnt; m++) { lc = &(hits->a[i].lc[gc->off + m]); fprintf(stderr, "*\tA\tutg%.6d%c\t%c\tqs:%u\tqe:%u\tql:%lu\tts:%u\tte:%u\ttl:%u\tcnt:%d\n", (lc->v>>1)+1, "lc"[ug->u.a[lc->v>>1].circ], "+-"[lc->v&1], lc->qs, lc->qe, hits->a[i].qlen, lc->ts, lc->te, ug->u.a[lc->v>>1].len, lc->cnt); } } } } void write_ul_hits(mg_gres_a *hits, mg_dbn_t *nn, const char *fn) { char *buf = (char*)calloc(strlen(fn) + 25, 1); sprintf(buf, "%s.ul.aln.bin", fn); FILE* fp = fopen(buf, "w"); uint32_t i; fwrite(&hits->n, sizeof(hits->n), 1, fp); for (i = 0; i < hits->n; i++) { fwrite(&hits->a[i].qid, sizeof(hits->a[i].qid), 1, fp); fwrite(&hits->a[i].qlen, sizeof(hits->a[i].qlen), 1, fp); fwrite(&hits->a[i].n_gc, sizeof(hits->a[i].n_gc), 1, fp); fwrite(&hits->a[i].n_lc, sizeof(hits->a[i].n_lc), 1, fp); fwrite(hits->a[i].gc, sizeof(mg_gchain_t), hits->a[i].n_gc, fp); fwrite(hits->a[i].lc, sizeof(mg_lres_t), hits->a[i].n_lc, fp); } // fwrite(hits->a, sizeof(mg_gres_t), hits->n, fp); fwrite(&hits->total_pair, sizeof(hits->total_pair), 1, fp); fwrite(&hits->total_base, sizeof(hits->total_base), 1, fp); fwrite(&(nn->n), sizeof(nn->n), 1, fp); fwrite(nn->a, sizeof(uint64_t), nn->n, fp); fwrite(&(nn->tl), sizeof(nn->tl), 1, fp); fwrite(&(nn->cc.n), sizeof(nn->cc.n), 1, fp); fwrite(nn->cc.a, sizeof(char), nn->cc.n, fp); // write_dbug(ug, fp); fclose(fp); fprintf(stderr, "[M::%s::] ==> UL alignments have been written\n", __func__); free(buf); } int load_ul_hits(mg_gres_a *hits, mg_dbn_t *nn, const char *fn) { uint64_t flag = 0; char *buf = (char*)calloc(strlen(fn) + 25, 1); sprintf(buf, "%s.ul.aln.bin", fn); FILE* fp = NULL; fp = fopen(buf, "r"); if(!fp) { free(buf); return 0; } uint32_t i; kv_init(*hits); flag += fread(&hits->n, sizeof(hits->n), 1, fp); hits->m = hits->n; MALLOC(hits->a, hits->n); for (i = 0; i < hits->n; i++) { flag += fread(&hits->a[i].qid, sizeof(hits->a[i].qid), 1, fp); flag += fread(&hits->a[i].qlen, sizeof(hits->a[i].qlen), 1, fp); flag += fread(&hits->a[i].n_gc, sizeof(hits->a[i].n_gc), 1, fp); flag += fread(&hits->a[i].n_lc, sizeof(hits->a[i].n_lc), 1, fp); MALLOC(hits->a[i].gc, hits->a[i].n_gc); MALLOC(hits->a[i].lc, hits->a[i].n_lc); flag += fread(hits->a[i].gc, sizeof(mg_gchain_t), hits->a[i].n_gc, fp); flag += fread(hits->a[i].lc, sizeof(mg_lres_t), hits->a[i].n_lc, fp); } // flag += fread(hits->a, sizeof(mg_gres_t), hits->n, fp); flag += fread(&hits->total_pair, sizeof(hits->total_pair), 1, fp); flag += fread(&hits->total_base, sizeof(hits->total_base), 1, fp); memset(nn, 0, sizeof(*nn)); flag += fread(&(nn->n), sizeof(nn->n), 1, fp); nn->m = nn->n; MALLOC(nn->a, nn->n); flag += fread(nn->a, sizeof(uint64_t), nn->n, fp); flag += fread(&(nn->tl), sizeof(nn->tl), 1, fp); flag += fread(&(nn->cc.n), sizeof(nn->cc.n), 1, fp); nn->cc.m = nn->cc.n; MALLOC(nn->cc.a, nn->cc.n); flag += fread(nn->cc.a, sizeof(char), nn->cc.n, fp); free(buf); // if(!test_dbug(ug, fp)) // { // free(hits->a.a); // kv_init(hits->a); // fclose(fp); // fprintf(stderr, "[M::%s::] ==> Renew Hi-C linkages\n", __func__); // return 0; // } fclose(fp); fprintf(stderr, "[M::%s::] ==> UL alignments have been loaded\n", __func__); return 1; } void get_asm_cov(ma_ug_t *ug, uint64_t ul_base, mul_ov_t *aov) { int64_t ss = asm_opt.hg_size; if(ss < 0) { uint64_t i, k, an; int64_t sp; asg_t *g = ug->g; asg_arc_t *av = NULL; for (i = 0, ss = 0; i < g->n_seq; i++) { sp = g->seq[i].len; av = asg_arc_a(g, i); an = asg_arc_n(g, i); for (k = 0; k < an; k++) { if(av[k].del) continue; if((av[k].v) < i) { sp -= ((int64_t)av[k].ol); } } if(sp > 0) ss += sp; } } else { ss *= asm_opt.polyploidy; } if(ss <= 0) ss = 1; aov->asm_cov = ul_base/ss; aov->asm_size = ss; fprintf(stderr, "[M::%s::] ==> asm_cov: %lu, asm_size: %lu\n", __func__, aov->asm_cov, aov->asm_size); } int32_t spec_ovlp_occ(eg_srt_t *a, int32_t a_n, int32_t st, int32_t vv, int32_t c_thres) { int32_t i, dst = a[st].d, occ = 1; if(occ >= c_thres) return 1; for (i = st + 1; i < a_n; i++) { if(a[i].id == a[st].id) continue; if(a[i].d - dst <= vv) { occ++; if(occ >= c_thres) return 1; } } for (i = st - 1; i >= 0; i--) { if(a[i].id == a[st].id) continue; if(dst - a[i].d <= vv) { occ++; if(occ >= c_thres) return 1; } } return 0; } int32_t get_spec_ovlp_occ(eg_srt_t *a, int32_t a_n, int32_t st, int32_t vv, int32_t c_thres, int32_t *s, int32_t *e, kvec_t_u64_warp *res) { int32_t i, dst = a[st].d, occ = 1, pp; (*s) = (*e) = st; res->a.n = 0; for (i = st + 1; i < a_n; i++) { if(a[i].d - dst <= vv) { (*e) = i; if(a[i].id == a[st].id) continue; occ++; kv_push(uint64_t, res->a, (((uint64_t)(a[i].id))<<32)|i); } else { break; } } for (i = st - 1; i >= 0; i--) { if(dst - a[i].d <= vv) { (*s) = i; if(a[i].id == a[st].id) continue; occ++; kv_push(uint64_t, res->a, (((uint64_t)(a[i].id))<<32)|i); } else { break; } } if(occ >= c_thres) { radix_sort_gfa64(res->a.a, res->a.a + res->a.n); for (i = 0, pp = -1, occ = 0; i < (int32_t)res->a.n; i++) { if((int32_t)(res->a.a[i]>>32) != pp) { pp = (res->a.a[i]>>32); res->a.a[occ] = res->a.a[i]; occ++; } } res->a.n = occ; if(occ >= c_thres) return occ; return 0; } else { return 0; } } void clean_ul_g(asg_t *xg) { uint32_t n_vtx = xg->n_seq * 2, v, i, nv, ie = 0, ike = 0; asg_arc_t *av = NULL; uint8_t* bs_flag = NULL; CALLOC(bs_flag, n_vtx); buf_t b; memset(&b, 0, sizeof(buf_t)); b.a = (binfo_t*)calloc(n_vtx, sizeof(binfo_t)); uint64_t max_dist = get_bub_pop_max_dist_advance(xg, &b); for (v = 0; v < xg->n_seq; v++) xg->seq[v].c = 0; for (v = 0; v < n_vtx; ++v) { if(bs_flag[v] != 0) continue; if (asg_arc_n(xg, v) < 2 || xg->seq[v>>1].del) continue; if(asg_bub_pop1_primary_trio(xg, NULL, v, max_dist, &b, (uint32_t)-1, (uint32_t)-1, 0, NULL, NULL, NULL, 0, 0, NULL)) { //beg is v, end is b.S.a[0] //note b.b include end, does not include beg for (i = 0; i < b.b.n; i++) { if(b.b.a[i]==v || b.b.a[i]==b.S.a[0]) continue; bs_flag[b.b.a[i]] = bs_flag[b.b.a[i]^1] = 1; } bs_flag[v] = 2; bs_flag[b.S.a[0]^1] = 3; } } for (v = 0; v < n_vtx; ++v) { if(bs_flag[v] != 0) continue; nv = asg_arc_n(xg, v); if (nv >= 2) { av = asg_arc_a(xg, v); for (i = 0; i < nv; ++i){ if (av[i].ol == 0) { av[i].del = 1; asg_arc_del(xg, av[i].v^1, (av[i].ul>>32)^1, 1); // fprintf(stderr, "---q0-utg%.6d%c, q1-utg%.6d%c\n", // (int32_t)((av[i].ul>>33)+1), "lc"[ug->u.a[av[i].ul>>33].circ], // (int32_t)((av[i].v)>>1)+1, "lc"[ug->u.a[av[i].v].circ]); } // fprintf(stderr, "xxxx-nv: %u, q0-utg%.6d%c, q1-utg%.6d%c\n", nv, // (int32_t)((av[i].ul>>33)+1), "lc"[ug->u.a[av[i].ul>>33].circ], // (int32_t)((av[i].v)>>1)+1, "lc"[ug->u.a[av[i].v].circ]); } } } for (i = 0; i < xg->n_arc; i++) { if(xg->arc[i].ol == 0) { ie++; if(!xg->arc[i].del) ike++; } } fprintf(stderr, "[M::%s::] ==> # fill gaps: %u, # keep gaps: %u\n", __func__, ie, ike); free(bs_flag); free(b.a); free(b.S.a); free(b.T.a); free(b.b.a); free(b.e.a); } // int32_t max_cluster(int32_t mmi, double vv, int32_t min_off, eg_srt_t *a, int32_t a_n, int32_t st, int32_t st_occ, int32_t *s, int32_t *e, kvec_t_u64_warp *res) // { // int32_t i, k, iocc, ovlp; // for (i = st, iocc = 0; i < k; i++) { // ovlp = (a[i].d > mmi? a[i].d - mmi: mmi - a[i].d) * vv; // if(ovlp < min_off) ovlp = min_off; // // fprintf(stderr, "i-%lu, ovlp: %d, td.a[i].d: %d, qid: %u\n", i, ovlp, td.a[i].d, td.a[i].id); // // if(spec_ovlp_occ(td.a + l, k-l, i - l, ovlp, c_thres)) break; // iocc = get_spec_ovlp_occ(td.a + l, k-l, i - l, ovlp, c_thres, &is, &ie, &tidx); // if(iocc >= c_thres) break; // } // } void get_ul_g(mul_ov_t *aov, mg_gres_a *hits, ma_ug_t *ug, const asg_t *rg, double cov_thres, double vv, int32_t min_off, int32_t min_read_ovlp) { int64_t c_thres = (aov->asm_cov*cov_thres)>2?(aov->asm_cov*cov_thres):2; uint64_t i, k, l, m, v0, v1, r0, r1; int32_t qs, qe, rs, re, qs0, qe0, qs1, qe1, ovlp, mmi, nngc2 = 0, is, ie, iocc, m_iocc, max_i; mg_gres_t *p = NULL; mg_gchain_t *gc = NULL, *gc0, *gc1; mg_lres_t *lf = NULL, *ll = NULL; asg_t *xg = copy_read_graph(ug->g); asg_arc_t *pe = NULL; kvec_t(lc_srt_t) tt; kv_init(tt); lc_srt_t *pt = NULL; kvec_t(eg_srt_t) td; kv_init(td); eg_srt_t *pd = NULL; kvec_t_u64_warp tidx; kv_init(tidx.a); ///for debug kvec_t(eg_srt_t) dbg_vw_srt; kv_init(dbg_vw_srt); for (i = 0; i < hits->n; i++) { // fprintf(stderr, "+i+: %lu\n",i); p = &(hits->a[i]); tt.n = 0; // fprintf(stderr, "-i-: %lu\n",i); if(p->n_gc < 2) continue; nngc2++; // fprintf(stderr, "\nsis: %lu, p->n_gc: %d\n",i,p->n_gc); for (k = 0; k < (uint64_t)p->n_gc; k++) { gc = &(p->gc[k]); assert(gc->cnt > 0); lf = &(p->lc[gc->off]); ll = gc->cnt>1?&(p->lc[gc->off+gc->cnt-1]):NULL; assert(lf->qs != (uint32_t)-1); if(ll) assert(ll->qs != (uint32_t)-1); transfor_icoord(lf->qs, lf->qe, lf->ts, lf->te, lf->v&1, p->qlen, ug->g->seq[lf->v>>1].len, &qs, ll?NULL:&qe, &rs, ll?NULL:&re); if(ll) { transfor_icoord(ll->qs, ll->qe, ll->ts, ll->te, ll->v&1, p->qlen, ug->g->seq[ll->v>>1].len, NULL, &qe, NULL, &re); } else { ll = lf; } if(qe - qs < min_read_ovlp || re - rs < min_read_ovlp) continue; kv_pushp(lc_srt_t, tt, &pt); pt->qse = qs; pt->qse <<= 32; pt->qse |= qe; pt->rse = rs; pt->rse <<= 32; pt->rse |= re; pt->gld = i; pt->gld <<= 32; pt->gld |= k; // fprintf(stderr, ">>>>k: %lu, qs: %d, qe: %d, qs-utg%.6d%c, qe-utg%.6d%c\n", k, qs, qe, // (int32_t)((lf->v>>1)+1), "lc"[ug->u.a[lf->v>>1].circ], // (int32_t)((ll->v>>1)+1), "lc"[ug->u.a[ll->v>>1].circ]); // fprintf(stderr, "lf_qs: %u, lf_qe: %u, lf_ts: %u, lf_te: %u\n", lf->qs, lf->qe, lf->ts, lf->te); // fprintf(stderr, "ll_qs: %u, ll_qe: %u, ll_ts: %u, ll_te: %u\n", ll->qs, ll->qe, ll->ts, ll->te); } // fprintf(stderr, "eie: %lu\n",i); radix_sort_lc_srt(tt.a, tt.a + tt.n); for (k = 0; k < tt.n; k++) { for (m = k + 1; m < tt.n; m++) { gc0 = &(p->gc[(uint32_t)(tt.a[k].gld)]); v0 = p->lc[gc0->off+gc0->cnt-1].v; gc1 = &(p->gc[(uint32_t)(tt.a[m].gld)]); v1 = p->lc[gc1->off].v; if((v0>>1) == (v1>>1)) continue; qs0 = tt.a[k].qse>>32; qe0 = (uint32_t)(tt.a[k].qse); qs1 = tt.a[m].qse>>32; qe1 = (uint32_t)(tt.a[m].qse); // fprintf(stderr, "++++k: %lu, qs0: %d, qe0: %d, qs1: %d, qe1: %d, q0-utg%.6d%c, q1-utg%.6d%c\n", // k, qs0, qe0, qs1, qe1, (int32_t)((v0>>1)+1), "lc"[ug->u.a[v0>>1].circ], (int32_t)((v1>>1)+1), "lc"[ug->u.a[v1>>1].circ]); if(qs1 <= qs0 && qe1 >= qe0) continue;///contain if(qs0 <= qs1 && qe0 >= qe1) continue;///contain if(ug->u.a[v0>>1].circ || ug->u.a[v1>>1].circ) continue; ovlp = ((MIN((qe0), (qe1)) > MAX((qs0), (qs1)))? MIN((qe0), (qe1)) - MAX((qs0), (qs1)):0); r0 = v0&1?(ug->u.a[v0>>1].start>>1):(ug->u.a[v0>>1].end>>1); r1 = v1&1?(ug->u.a[v1>>1].end>>1):(ug->u.a[v1>>1].start>>1); // fprintf(stderr, "----k: %lu, ovlp: %d\n", k, ovlp); // if((ovlp == 0) || (ovlp <= ((qe0 - qs0)*vv) && ovlp <= ((qe1 - qs1)*vv)) || // (asg_arc_n(ug->g, v0) == 0 && asg_arc_n(ug->g, v1^1) == 0)) { if(/**(asg_arc_n(ug->g, v0) == 0 && asg_arc_n(ug->g, v1^1) == 0) && **/(ovlp < (int32_t)(MIN(rg->seq[r0].len, rg->seq[r1].len)))) { kv_pushp(eg_srt_t, td, &pd); pd->d = MAX((qs0), (qs1)) - MIN((qe0), (qe1)); pd->x = v0id = p->qid; pd->e = (uint32_t)(tt.a[k].gld); pd->e <<= 32; pd->e |= (uint32_t)(tt.a[m].gld); } } } } fprintf(stderr, "td.n: %d\n", (int)td.n); radix_sort_eg_srt_x(td.a, td.a + td.n); for (k = 1, l = 0; k <= td.n; ++k) { if (k == td.n || td.a[k].x != td.a[l].x) { if(k - l >= (uint64_t)c_thres) { for (i = l+1, mmi = l; i < k; i++) { if(td.a[mmi].d > td.a[i].d) mmi = i; } mmi = td.a[mmi].d < 0? -td.a[mmi].d:0; if(mmi != 0) { for (i = l; i < k; i++) td.a[i].d += mmi; } radix_sort_eg_srt_d(td.a + l, td.a + k); for (i = l, iocc = 0, tidx.a.n = 0; i < k; i++) { ovlp = (td.a[i].d > mmi? td.a[i].d - mmi: mmi - td.a[i].d) * vv; if(ovlp < min_off) ovlp = min_off; // fprintf(stderr, "i-%lu, ovlp: %d, td.a[i].d: %d, qid: %u\n", i, ovlp, td.a[i].d, td.a[i].id); // if(spec_ovlp_occ(td.a + l, k-l, i - l, ovlp, c_thres)) break; iocc = get_spec_ovlp_occ(td.a + l, k-l, i - l, ovlp, c_thres, &is, &ie, &tidx); // fprintf(stderr, "c_thres-%ld, iocc-%d\n", c_thres, iocc); if(iocc >= c_thres) break; } if(i < k) { m_iocc = iocc; max_i = i; for (i = ie + 1; i < k; i++) { iocc = get_spec_ovlp_occ(td.a + l, k-l, i - l, ovlp, m_iocc, &is, &ie, &tidx); if(iocc > m_iocc) m_iocc = iocc, max_i = i; i = ie + l; } ///for debug kv_pushp(eg_srt_t, dbg_vw_srt, &pd); pd->x = m_iocc; pd->e = td.a[l].x; v0 = (uint32_t)td.a[l].x; v1 = td.a[l].x>>32; pe = asg_arc_pushp(xg); pe->del = 0; pe->strong = 0; pe->el = 0; pe->no_l_indel = 0; pe->ol = 0; pe->v = v0; pe->ul = v1<<32; pe->ul += xg->seq[v1>>1].len; v0 = (td.a[l].x>>32)^1; v1 = ((uint32_t)td.a[l].x)^1; pe = asg_arc_pushp(xg); pe->del = 0; pe->strong = 0; pe->el = 0; pe->no_l_indel = 0; pe->ol = 0; pe->v = v0; pe->ul = v1<<32; pe->ul += xg->seq[v1>>1].len; // fprintf(stderr, "++++q0-utg%.6d%c, q1-utg%.6d%c, k-l: %lu, c_thres: %ld, flag: %u\n", // (int32_t)((td.a[l].x>>33)+1), "lc"[ug->u.a[td.a[l].x>>33].circ], // (int32_t)(((uint32_t)td.a[l].x)>>1)+1, "lc"[ug->u.a[(((uint32_t)td.a[l].x)>>1)].circ], k-l, c_thres, // (asg_arc_n(ug->g, ((uint32_t)td.a[l].x)^1) == 0 && asg_arc_n(ug->g, (td.a[l].x>>32)) == 0)); } } l = k; } } xg->is_srt = 0; xg->idx = 0; free(xg->idx); asg_cleanup(xg); clean_ul_g(xg); ///for debug fprintf(stderr, "[M::%s::] ==> nngc2: %d\n", __func__, nngc2); radix_sort_eg_srt_x(dbg_vw_srt.a, dbg_vw_srt.a + dbg_vw_srt.n); for (max_i = (int32_t)dbg_vw_srt.n - 1; max_i >= 0; --max_i) { pd = &(dbg_vw_srt.a[max_i]); fprintf(stderr, "++++q0-utg%.6d%c, q1-utg%.6d%c, occ: %lu, c_thres: %ld, flag: %u\n", (int32_t)((pd->e>>33)+1), "lc"[ug->u.a[pd->e>>33].circ], (int32_t)(((uint32_t)pd->e)>>1)+1, "lc"[ug->u.a[(((uint32_t)pd->e)>>1)].circ], pd->x, c_thres, (asg_arc_n(ug->g, ((uint32_t)pd->e)^1) == 0 && asg_arc_n(ug->g, (pd->e>>32)) == 0)); } kv_destroy(tt); kv_destroy(td); kv_destroy(tidx.a); kv_destroy(dbg_vw_srt); asg_destroy(xg); } int ul_align(mg_idxopt_t *opt, const ug_opt_t *uopt, const asg_t *rg, const enzyme *fn, void *ha_flt_tab, ha_pt_t *ha_idx, ma_ug_t *ug) { uldat_t sl; memset(&sl, 0, sizeof(sl)); sl.ha_flt_tab = ha_flt_tab; sl.ha_idx = ha_idx; sl.opt = opt; sl.chunk_size = 200000000; sl.n_thread = asm_opt.thread_num; sl.ug = ug; sl.rg = rg; sl.uopt = uopt; if(!load_ul_hits(&sl.hits, &sl.nn, asm_opt.output_file_name)) { alignment_ul_pipeline(&sl, fn); write_ul_hits(&sl.hits, &sl.nn, asm_opt.output_file_name); } mul_ov_t aov; memset(&aov, 0, sizeof(aov)); get_asm_cov(ug, sl.hits.total_base, &aov); fprintf(stderr, "[M::%s::] ==> total_pair: %lu, total_base: %lu, n: %d\n", __func__, sl.hits.total_pair, sl.hits.total_base, (int32_t)sl.hits.n); get_ul_g(&aov, &sl.hits, ug, rg, 0.51, 0.1, 500, 1000); // print_gaf(ug, &(sl.hits), &(sl.nn)); mg_gres_a_des(&(sl.hits)); free(sl.nn.a); free(sl.nn.cc.a); return 1; } void ul_resolve(ma_ug_t *ug, const asg_t *rg, const ug_opt_t *uopt, int hap_n) { fprintf(stderr, "[M::%s::] ==> UL\n", __func__); mg_idxopt_t opt; init_mg_opt(&opt, 0, 19, 10, hap_n, 0, 0, 0.05); int exist = (asm_opt.load_index_from_disk? uidx_load(&ha_flt_tab, &ha_idx, asm_opt.output_file_name) : 0); if(exist == 0) uidx_build(ug, &opt); if(exist == 0) uidx_write(ha_flt_tab, ha_idx, asm_opt.output_file_name); ul_align(&opt, uopt, rg, asm_opt.ar, ha_flt_tab, ha_idx, ug); uidx_destory(); } void ul_v_call(uldat_t *sl, const enzyme *fn) { scall_ul_pipeline(sl, fn); // UL_INF; // print_ul_rs(&UL_INF); // debug_retrieve_rc_sub(uopt, &UL_INF, &R_INF, (ul_idx_t *)sl.uu, 100); // if(!load_ul_hits(&sl.hits, &sl.nn, asm_opt.output_file_name)) { // scall_ul_pipeline(&sl, fn); // write_ul_hits(&sl.hits, &sl.nn, asm_opt.output_file_name); // } } void print_dedup_HiFis_seq(ma_ug_t *ug) { uint64_t i; ma_utg_t *p = NULL; for (i = 0; i < ug->u.n; i++) { p = &(ug->u.a[i]); CALLOC(p->s, p->len+1); retrieve_u_seq(NULL, p->s, p, 0, 0, -1, NULL); p->s[p->len] = '\0'; } FILE* output_file = fopen("dedup_HiFis_seq.gfa", "w"); ma_ug_print(ug, NULL, NULL, NULL, NULL, "utg", output_file); fclose(output_file); output_file = fopen("dedup_HiFis_seq.noseq.gfa", "w"); ma_ug_print_simple(ug, NULL, NULL, NULL, NULL, "utg", output_file); fclose(output_file); exit(1); } void push_coverage_track(ucov_t *cc, ul_contain *ct, uint64_t uid, ma_utg_t *u, asg_t *rg, ma_hit_t_alloc* src, int64_t min_ovlp, int64_t max_hang, uint64_t is_el, uint64_t is_del) { uint64_t k, l, z, dp, ct_n; utg_ct_t *ct_a = NULL; cc->idx[uid] = cc->interval.n; ct_n = ((uint32_t)(ct->idx.a[uid])); ct_a = ct->rids.a + ((ct->idx.a[uid])>>32); for (z = 0; z < ct_n; z++) { kv_push(uint64_t, cc->interval, ct_a[z].s<<1); kv_push(uint64_t, cc->interval, (ct_a[z].e<<1)|1); } for (k = l = 0; k < u->n; k++) { kv_push(uint64_t, cc->interval, l<<1); kv_push(uint64_t, cc->interval, ((l + Get_READ_LENGTH(R_INF, u->a[k]>>33))<<1)|1); /** i = u->a[k]>>33;///rid for (z = 0; z < src[i].length; z++) { if(is_el && (!src[i].buffer[z].el)) continue; if(is_del && (!src[i].buffer[z].del)) continue; qn = Get_qn(src[i].buffer[z]); tn = Get_tn(src[i].buffer[z]); if(!rg->seq[tn].del) continue; if((Get_qe(src[i].buffer[z]) - Get_qs(src[i].buffer[z])) < min_ovlp) continue; if((Get_te(src[i].buffer[z]) - Get_ts(src[i].buffer[z])) < min_ovlp) continue; r = ma_hit2arc(&(src[i].buffer[z]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r != MA_HT_TCONT) continue;///tn is contained ori = (u->a[k]>>32)&1; if(ori == 0) { qs = Get_qs(src[i].buffer[z]); qe = Get_qe(src[i].buffer[z]); } else { qs = (Get_READ_LENGTH(R_INF, i)) - Get_qe(src[i].buffer[z]); qe = (Get_READ_LENGTH(R_INF, i)) - Get_qs(src[i].buffer[z]); } kv_push(uint64_t, cc->interval, (l+qs)<<1); kv_push(uint64_t, cc->interval, ((l+qe)<<1)|1); } **/ l += (uint32_t)u->a[k]; } cc->idx[uid+1] = cc->interval.n; radix_sort_gfa64(cc->interval.a+cc->idx[uid], cc->interval.a+cc->interval.n); for (k = cc->idx[uid], dp = 0; k < cc->interval.n; ++k) { ///if a[j] is qe if (cc->interval.a[k]&1) --dp; else ++dp; l = cc->interval.a[k]>>1; l <<= 32; l += dp; cc->interval.a[k] = l; } } uint32_t check_if_fully_contain(uint32_t sid, uint32_t lid, uint32_t ori, uint8_t *rset, asg_t *rg, ma_hit_t_alloc* src, int64_t min_ovlp, int64_t max_hang, int64_t gap_fuzz) { uint32_t rid, k, qn, tn, ff = 1; int32_t r; asg_arc_t t; return 1; rid = lid; for (k = 0; k < src[rid].length; k++) { if(!src[rid].buffer[k].el) continue; qn = Get_qn(src[rid].buffer[k]); tn = Get_tn(src[rid].buffer[k]); if(rg->seq[qn].del || rg->seq[tn].del) continue; if((Get_qe(src[rid].buffer[k]) - Get_qs(src[rid].buffer[k])) < min_ovlp) continue; if((Get_te(src[rid].buffer[k]) - Get_ts(src[rid].buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(src[rid].buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0) continue; rset[t.v] = ((t.ul>>32)&1)+1; } rid = sid; for (k = 0; k < src[rid].length; k++) { if(!src[rid].buffer[k].el) continue; qn = Get_qn(src[rid].buffer[k]); tn = Get_tn(src[rid].buffer[k]); if(rg->seq[qn].del || rg->seq[tn].del) continue; if((Get_qe(src[rid].buffer[k]) - Get_qs(src[rid].buffer[k])) < min_ovlp) continue; if((Get_te(src[rid].buffer[k]) - Get_ts(src[rid].buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(src[rid].buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0) continue; if(rset[t.v] != ((((t.ul>>32)&1)^ori)+1)) { ff = 0; break; } } rid = lid; for (k = 0; k < src[rid].length; k++) { if(!src[rid].buffer[k].el) continue; qn = Get_qn(src[rid].buffer[k]); tn = Get_tn(src[rid].buffer[k]); if(rg->seq[qn].del || rg->seq[tn].del) continue; if((Get_qe(src[rid].buffer[k]) - Get_qs(src[rid].buffer[k])) < min_ovlp) continue; if((Get_te(src[rid].buffer[k]) - Get_ts(src[rid].buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(src[rid].buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0) continue; rset[t.v] = 0; } return ff; } ul_contain *ul_contain_gen(ma_ug_t *ug, asg_t *rg, ma_hit_t_alloc* src, int64_t min_ovlp, int64_t max_hang, uint64_t is_el, uint64_t is_del) { uint64_t k, l, i, z, t, qn, tn, ori, qs, qe, ovlp, o_z, o_r, o_o; ul_contain *p = NULL; ma_utg_t *u = NULL; utg_ct_t *m = NULL; int32_t r; asg_arc_t e; CALLOC(p, 1); p->idx.n = p->idx.m = ug->u.n; CALLOC(p->idx.a, p->idx.n); p->is_c.n = rg->n_seq; CALLOC(p->is_c.a, p->is_c.n); for (t = 0; t < ug->u.n; t++) { u = &(ug->u.a[t]); p->idx.a[t] = p->rids.n; p->idx.a[t] <<= 32; for (k = l = 0; k < u->n; k++) { i = u->a[k]>>33;///rid for (z = 0; z < src[i].length; z++) { if(is_el && (!src[i].buffer[z].el)) continue; if(is_del && (!src[i].buffer[z].del)) continue; qn = Get_qn(src[i].buffer[z]); tn = Get_tn(src[i].buffer[z]); if(!rg->seq[tn].del) continue; if((Get_qe(src[i].buffer[z]) - Get_qs(src[i].buffer[z])) < min_ovlp) continue; if((Get_te(src[i].buffer[z]) - Get_ts(src[i].buffer[z])) < min_ovlp) continue; r = ma_hit2arc(&(src[i].buffer[z]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &e); if(r != MA_HT_TCONT) continue;///tn is contained p->is_c.a[qn] = 1; ori = (u->a[k]>>32)&1; if(ori == 0) { qs = Get_qs(src[i].buffer[z]); qe = Get_qe(src[i].buffer[z]); } else { qs = (Get_READ_LENGTH(R_INF, i)) - Get_qe(src[i].buffer[z]); qe = (Get_READ_LENGTH(R_INF, i)) - Get_qs(src[i].buffer[z]); } qs += l; qe += l; kv_pushp(utg_ct_t, p->rids, &m); m->x = tn; m->x <<= 1; m->x |= (src[i].buffer[z].rev == ori?0:1); m->s = qs; m->e = qe/** + 1**/; } l += (uint32_t)u->a[k]; } radix_sort_utg_ct_t_x_srt(p->rids.a + (p->idx.a[t]>>32), p->rids.a + p->rids.n); /** for (k = (p->idx.a[t]>>32) + 1, l = i = (p->idx.a[t]>>32); k <= p->rids.n; ++k) { if (k == p->rids.n || (p->rids.a[k].x>>1) != (p->rids.a[l].x>>1)) { p->rids.a[i] = p->rids.a[l]; i++; l = k; } } **/ for (k = (p->idx.a[t]>>32) + 1, l = i = (p->idx.a[t]>>32); k <= p->rids.n; ++k) { if (k == p->rids.n || p->rids.a[k].x != p->rids.a[l].x) { for (z = l; z < k; z++) { for (r = (int64_t)i-1; r >= 0 && p->rids.a[r].x == p->rids.a[z].x; r--) { ovlp = ((MIN(p->rids.a[z].e, p->rids.a[r].e) > MAX(p->rids.a[z].s, p->rids.a[r].s))? (MIN(p->rids.a[z].e, p->rids.a[r].e) - MAX(p->rids.a[z].s, p->rids.a[r].s)):0); if(ovlp) { o_z = p->rids.a[z].e - p->rids.a[z].s; o_r = p->rids.a[r].e - p->rids.a[r].s; o_o = MIN(o_z, o_r); if((ovlp <= o_o*1.05) && (ovlp >= o_o*0.95)) break; } // if(p->rids.a[z].s == p->rids.a[r].s && p->rids.a[z].e == p->rids.a[r].e) break; } if(r >= 0 && p->rids.a[r].x == p->rids.a[z].x) continue; p->rids.a[i++] = p->rids.a[z]; } l = k; } } p->rids.n = i; radix_sort_utg_ct_t_s_srt(p->rids.a + (p->idx.a[t]>>32), p->rids.a + p->rids.n); p->idx.a[t] |= (p->rids.n - (p->idx.a[t]>>32)); } // fprintf(stderr, "p->rids.n:%u, p->idx.n:%u\n", (uint32_t)p->rids.n, (uint32_t)p->idx.n); return p; } void debug_append_inexact_edges(ma_ug_t *ug, const ug_opt_t *uopt) { uint32_t n_asymm = 0, n_disconnect = 0, z, v, w, k, nv; asg_arc_t *av = NULL; for (z = 0; z < ug->g->n_arc; ++z) { if(ug->g->arc[z].del) continue; if(!get_ug_edge_src(ug, uopt->sources, uopt->max_hang, uopt->min_ovlp, ug->g->arc[z].ul>>32, ug->g->arc[z].v)) { n_disconnect++; } v = ug->g->arc[z].v^1; w = (ug->g->arc[z].ul>>32)^1; nv = asg_arc_n(ug->g, v); av = asg_arc_a(ug->g, v); for (k = 0; k < nv; ++k) { if (av[k].del) continue; // fprintf(stderr, "found <%lu> -> <%u>\n", av[k].ul>>32, av[k].v); if (av[k].v == w) break; } if (k == nv) { ug->g->arc[z].del = 1, ++n_asymm; // fprintf(stderr, "# lack of <%u> -> <%u>, should be <%u> -> <%u>\n\n", w^1, v^1, v, w); } } if(n_asymm || n_disconnect) { asg_cleanup(ug->g); fprintf(stderr, "[M::%s] # asymm edges: %u, # disconnect edges: %u\n", __func__, n_asymm, n_disconnect); // exit(1); } } void append_inexact_edges(ma_ug_t *ug, const ug_opt_t *uopt, asg_t *rg) { uint32_t *idx = NULL, n_read = R_INF.total_reads, z, v, k, qn, tn, tu, ut_v, ut_w; ma_utg_t *u = NULL; ma_hit_t_alloc *src = uopt->sources, *s = NULL; int32_t r; asg_arc_t t, *p = NULL; int64_t min_ovlp = uopt->min_ovlp, max_hang = uopt->max_hang, occ = 0; MALLOC(idx, n_read); memset(idx, -1, n_read*sizeof(*(idx))); for (z = 0; z < ug->u.n; z++) { u = &(ug->u.a[z]); if(u->circ) continue; idx[u->start>>1] = idx[u->end>>1] = z; } for (z = 0; z < ug->u.n; z++) { u = &(ug->u.a[z]); if(u->circ) continue; v = u->end^1; s = &(src[v>>1]); ut_v = (z<<1); for (k = 0; k < s->length; k++) { if(s->buffer[k].el) continue;///we just need inexact edges qn = Get_qn(s->buffer[k]); tn = Get_tn(s->buffer[k]); tu = idx[tn]; ut_w = (uint32_t)-1; if(tu == (uint32_t)-1 || ug->g->seq[tu].del) continue; if((Get_qe(s->buffer[k]) - Get_qs(s->buffer[k])) < min_ovlp) continue; if((Get_te(s->buffer[k]) - Get_ts(s->buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(s->buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0 || (t.ul>>32) != v) continue; if(t.v == ug->u.a[tu].start) ut_w = tu<<1; if(t.v == ug->u.a[tu].end) ut_w = (tu<<1)+1; if(ut_w==(uint32_t)-1) continue; p = asg_arc_pushp(ug->g); memset(p, 0, sizeof(*p)); *p = t; p->ul = ut_v; p->ul <<= 32; p->ul += ((uint32_t)(t.ul)); p->v = ut_w; occ++; // if((p->v>>1)>=ug->g->n_seq || (p->ul>>33)>=ug->g->n_seq) { // fprintf(stderr, "+ug->g->n_seq:%u, (p->ul>>33):%u, (p->v>>1):%u\n", // (uint32_t)ug->g->n_seq, (uint32_t)(p->ul>>33), (uint32_t)(p->v>>1)); // } // assert((p->v>>1)g->n_seq && (p->ul>>33)g->n_seq); } v = u->start^1; s = &(src[v>>1]); ut_v = (z<<1) + 1; for (k = 0; k < s->length; k++) { if(s->buffer[k].el) continue;///we just need inexact edges qn = Get_qn(s->buffer[k]); tn = Get_tn(s->buffer[k]); tu = idx[tn]; ut_w = (uint32_t)-1; if(tu == (uint32_t)-1 || ug->g->seq[tu].del) continue; if((Get_qe(s->buffer[k]) - Get_qs(s->buffer[k])) < min_ovlp) continue; if((Get_te(s->buffer[k]) - Get_ts(s->buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(s->buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if(r < 0 || (t.ul>>32) != v) continue; if(t.v == ug->u.a[tu].start) ut_w = tu<<1; if(t.v == ug->u.a[tu].end) ut_w = (tu<<1)+1; if(ut_w==(uint32_t)-1) continue; p = asg_arc_pushp(ug->g); memset(p, 0, sizeof(*p)); *p = t; p->ul = ut_v; p->ul <<= 32; p->ul += ((uint32_t)(t.ul)); p->v = ut_w; occ++; // if((p->v>>1)>=ug->g->n_seq || (p->ul>>33)>=ug->g->n_seq) { // fprintf(stderr, "+ug->g->n_seq:%u, (p->ul>>33):%u, (p->v>>1):%u\n", // (uint32_t)ug->g->n_seq, (uint32_t)(p->ul>>33), (uint32_t)(p->v>>1)); // } // assert((p->v>>1)g->n_seq && (p->ul>>33)g->n_seq); } } if(occ) { free(ug->g->idx); ug->g->idx = 0; ug->g->is_srt = 0; asg_cleanup(ug->g); } free(idx); ///for debug debug_append_inexact_edges(ug, uopt); fprintf(stderr, "[M::%s] # inserted inexact edges: %ld\n", __func__, occ); } typedef struct { ucov_t *cr; ma_hit_t_alloc* src; int64_t min_ovlp; int64_t max_hang; uint64_t is_el; uint64_t is_del; uint64_t is_src_cc; asg_t *rg; ma_ug_t *ug; } r_contain_aux; static void update_gen_r_contain(void *data, long i, int tid) // callback for kt_for() { r_contain_aux *s = (r_contain_aux *)data; ma_hit_t_alloc *src = s->src; ma_hit_t *t; int32_t r; asg_arc_t x; uint64_t *a = s->cr->interval.a + s->cr->idx[i], a_n = s->cr->idx[i+1] - s->cr->idx[i], k, dp, l, z, qn, tn; uint64_t is_el = s->is_el, is_del = s->is_del, min_ovlp = s->min_ovlp, max_hang = s->max_hang, qs, qe, cs, ce, sum; int64_t ii; asg_t *rg = s->rg; uint64_t *b, b_n, ti; // if(a_n == 0 || rg->seq[i].del) return; if(s->is_src_cc) { for (z = 0; z < src[i].length; z++) { t = &(src[i].buffer[z]); t->cc = 0; if(a_n == 0 || rg->seq[i].del) continue; qn = Get_qn((*t)); tn = Get_tn((*t)); if(qn > tn) continue; if(is_el && (!(t->el))) continue; if(is_del && (!(t->del))) continue; if((Get_qe((*t)) - Get_qs((*t))) < min_ovlp) continue; if((Get_te((*t)) - Get_ts((*t))) < min_ovlp) continue; if(rg->seq[tn].del) continue; b = s->cr->interval.a + s->cr->idx[tn]; b_n = s->cr->idx[tn+1] - s->cr->idx[tn]; if(b_n == 0) continue; r = ma_hit2arc(t, Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &x); if(r < 0) continue; qs = Get_qs((*t)); qe = Get_qe((*t)); for (k = 0; k < a_n; k += 2) { cs = a[k]>>33; ce = a[k+1]>>33; assert(rg->seq[(uint32_t)(a[k])].del); if(qs<=cs+128 && qe+128>=ce) { ///128 is the offset for indel for (ti = 0; ti < b_n; ti+=2) { if((uint32_t)(b[ti]) == (uint32_t)(a[k])) { sum = t->cc; sum += (ce - cs); if(sum > 0x3fffffffU) sum = 0x3fffffffU; t->cc = sum; break; } } } } } } else { radix_sort_gfa64(a, a + a_n); for (k = 0, dp = 0; k < a_n; ++k) { ///if a[j] is qe if ((a[k]>>32)&1) --dp; else ++dp; l = a[k]>>33; l <<= 32; l += dp; a[k] = l; } for (z = 0; z < src[i].length; z++) { t = &(src[i].buffer[z]); qn = Get_qn((*t)); tn = Get_tn((*t)); if(qn > tn) continue; if(t->cc == 0) continue; ii = get_specific_overlap(&(src[tn]), tn, qn); src[tn].buffer[ii].cc = t->cc; } } } static void update_ug_uo_t(void *data, long i, int tid) { r_contain_aux *sl = (r_contain_aux *)data; int32_t r; ma_hit_t_alloc *src = sl->src, *x; uint32_t k, qn, tn, uv, uw, v, w; asg_arc_t *e = &(sl->ug->g->arc[i]), t; uv = e->ul>>32; uw = e->v; e->ou = 0; if(sl->ug->u.a[uv>>1].circ || sl->ug->u.a[uw>>1].circ) return; v = ((uv&1)?(sl->ug->u.a[uv>>1].start^1):(sl->ug->u.a[uv>>1].end^1)); w = ((uw&1)?(sl->ug->u.a[uw>>1].end):(sl->ug->u.a[uw>>1].start)); x = &(src[v>>1]); for (k = 0; k < x->length; k++) { qn = Get_qn(x->buffer[k]); tn = Get_tn(x->buffer[k]); if(qn == (v>>1) && tn == (w>>1)) { r = ma_hit2arc(&(x->buffer[k]), sl->rg->seq[v>>1].len, sl->rg->seq[w>>1].len, sl->max_hang, asm_opt.max_hang_rate, sl->min_ovlp, &t); if(r < 0) continue; if((t.ul>>32)!=v || t.v!=w) continue; e->ou = (x->buffer[k].cc>OU_MASK?OU_MASK:x->buffer[k].cc); break; } } assert(k < x->length); } ucov_t *gen_r_contain(ma_ug_t *ug, asg_t *rg, ma_hit_t_alloc* src, uint64_t n_read, int64_t min_ovlp, int64_t max_hang, uint64_t n_thread, uint64_t is_el, uint64_t is_del) { ucov_t *cr = NULL; uint64_t i, z, qn, tn, qs, qe; ma_hit_t *t = NULL; int32_t r; asg_arc_t x; CALLOC(cr, 1); MALLOC(cr->idx, n_read+1); kv_init(cr->interval); for (i = 0; i < n_read; i++) { cr->idx[i] = cr->interval.n; if(rg->seq[i].del) continue; for (z = 0; z < src[i].length; z++) { t = &(src[i].buffer[z]); t->cc = 0; if(is_el && (!(t->el))) continue; if(is_del && (!(t->del))) continue; if((Get_qe((*t)) - Get_qs((*t))) < min_ovlp) continue; if((Get_te((*t)) - Get_ts((*t))) < min_ovlp) continue; qn = Get_qn((*t)); tn = Get_tn((*t)); if(!rg->seq[tn].del) continue; r = ma_hit2arc(t, Get_READ_LENGTH(R_INF, qn), Get_READ_LENGTH(R_INF, tn), max_hang, asm_opt.max_hang_rate, min_ovlp, &x); if(r == MA_HT_TCONT) { ///tn is contained qs = Get_qs((*t)); qe = Get_qe((*t)); kv_push(uint64_t, cr->interval, ((qs<<1)<<32)|tn); kv_push(uint64_t, cr->interval, (((qe<<1)|1)<<32)|tn); } } } cr->idx[i] = cr->interval.n; // fprintf(stderr, "+++[M::%s]n_read:%lu\n", __func__, n_read); r_contain_aux aux; aux.cr = cr; aux.src = src; aux.min_ovlp = min_ovlp; aux.rg = rg; aux.ug = ug; aux.max_hang = max_hang; aux.is_el = 0/**is_el**/; aux.is_del = 0/**is_del**/; aux.is_src_cc = 1; kt_for(n_thread, update_gen_r_contain, &aux, n_read);///note: here we should set is_el = is_del = 0 aux.is_src_cc = 0; kt_for(n_thread, update_gen_r_contain, &aux, n_read); if(ug) kt_for(n_thread, update_ug_uo_t, &aux, ug->g->n_arc); return cr; } ucov_t *gen_cov_track(ma_ug_t *ug, asg_t *rg, ul_contain *ct, ma_hit_t_alloc* src, int64_t min_ovlp, int64_t max_hang, uint64_t is_el, uint64_t is_del) { uint64_t i, k; ucov_t *cc = NULL; CALLOC(cc, 1); MALLOC(cc->idx, ug->u.n+1); kv_init(cc->interval); for (i = k = 0; i < ug->u.n; i++) { k += ug->u.a[i].len; push_coverage_track(cc, ct, i, &(ug->u.a[i]), rg, src, min_ovlp, max_hang, is_el, is_del); } fprintf(stderr, "[M::%s::] # bases: %lu\n", __func__, k); return cc; } ul_idx_t *dedup_HiFis(const ug_opt_t *uopt, uint64_t is_el, uint64_t is_del) { uint64_t i, k, qn, tn, n_read = R_INF.total_reads, cc_num = 0; int32_t r; asg_arc_t t, *p = NULL; uint8_t *rset = NULL; CALLOC(rset, n_read<<1); asg_t *rg = asg_init(); ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang; int64_t gap_fuzz = uopt->gap_fuzz; rg->m_seq = rg->n_seq = n_read; MALLOC(rg->seq, rg->m_seq); for (i = 0; i < n_read; ++i) { rg->seq[i].len = Get_READ_LENGTH(R_INF, i); rg->seq[i].del = rg->seq[i].c = 0; } for (i = 0; i < n_read; i++) { if(rg->seq[i].del) continue; for (k = 0; k < src[i].length; k++) { if(is_el && (!src[i].buffer[k].el)) continue; if(is_del && (!src[i].buffer[k].del)) continue; qn = Get_qn(src[i].buffer[k]); tn = Get_tn(src[i].buffer[k]); if(rg->seq[qn].del || rg->seq[tn].del) continue; if((Get_qe(src[i].buffer[k]) - Get_qs(src[i].buffer[k])) < min_ovlp) continue; if((Get_te(src[i].buffer[k]) - Get_ts(src[i].buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(src[i].buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if (r == MA_HT_QCONT/** && check_if_fully_contain(qn, tn, src[i].buffer[k].rev, rset, rg, src, min_ovlp, max_hang, gap_fuzz)**/) { rg->seq[qn].del = 1; } else if(r == MA_HT_TCONT/** && check_if_fully_contain(tn, qn, src[i].buffer[k].rev, rset, rg, src, min_ovlp, max_hang, gap_fuzz)**/) { rg->seq[tn].del = 1; } if(rg->seq[i].del) break; } } for (i = 0; i < n_read; i++) { if(rg->seq[i].del) {cc_num++; continue;} for (k = 0; k < src[i].length; k++) { if(is_el && (!src[i].buffer[k].el)) continue; if(is_del && (!src[i].buffer[k].del)) continue; qn = Get_qn(src[i].buffer[k]); tn = Get_tn(src[i].buffer[k]); if(rg->seq[qn].del || rg->seq[tn].del) continue; if((Get_qe(src[i].buffer[k]) - Get_qs(src[i].buffer[k])) < min_ovlp) continue; if((Get_te(src[i].buffer[k]) - Get_ts(src[i].buffer[k])) < min_ovlp) continue; r = ma_hit2arc(&(src[i].buffer[k]), rg->seq[qn].len, rg->seq[tn].len, max_hang, asm_opt.max_hang_rate, min_ovlp, &t); if (r >= 0) { p = asg_arc_pushp(rg); *p = t; } } } asg_cleanup(rg); asg_symm(rg); asg_arc_del_trans(rg, gap_fuzz); ma_ug_t *ug = NULL; ug = ma_ug_gen(rg); append_inexact_edges(ug, uopt, rg); ul_idx_t *uu = NULL; CALLOC(uu, 1); uu->ug = ug; uu->ct = ul_contain_gen(ug, rg, src, min_ovlp, max_hang, is_el, is_del); uu->cc = gen_cov_track(ug, rg, uu->ct, src, min_ovlp, max_hang, is_el, is_del); uu->cr = gen_r_contain(ug, rg, src, n_read, min_ovlp, max_hang, asm_opt.thread_num, is_el, is_del); // uu->ov = compress_dedup_HiFis(ug, src); asg_destroy(rg); free(rset); // uu->nug = cvert_t_gen(uopt); fprintf(stderr, "[M::%s::] # unitigs: %lu, # edges: %lu, # cc_num: %lu\n", __func__, (uint64_t)ug->u.n, (uint64_t)ug->g->n_arc, cc_num); // print_dedup_HiFis_seq(ug); return uu; } utg_rid_t *gen_r_ug_idx(ma_ug_t *ug, asg_t *rg) { uint64_t i, k, l, m, rid, a_n; utg_rid_dt *a; ma_utg_t *u = NULL; utg_rid_t *cc = NULL; CALLOC(cc, 1); CALLOC(cc->idx, rg->n_seq+1); kv_init(cc->p); cc->rg = rg; for (i = 0; i < ug->u.n; i++) { u = &(ug->u.a[i]); for (k = 0; k < u->n; k++) cc->idx[u->a[k]>>33]++; } for (k = l = 0; k <= rg->n_seq; k++) { m = cc->idx[k]; cc->idx[k] = l; l += m; } cc->p.n = cc->p.m = l; CALLOC(cc->p.a, cc->p.n); for (i = 0; i < ug->u.n; i++) { u = &(ug->u.a[i]); for (k = l = 0; k < u->n; k++) { rid = u->a[k]>>33; a = cc->p.a + cc->idx[rid]; a_n = cc->idx[rid+1] - cc->idx[rid]; if(a_n) { if(a[a_n-1].off == a_n-1) { a[a_n-1].u = (i<<1)|((u->a[k]>>32)&1); a[a_n-1].pos = l; a[a_n-1].off = l; } else { a[a[a_n-1].off].u = (i<<1)|((u->a[k]>>32)&1); a[a[a_n-1].off].pos = l; a[a[a_n-1].off].off = l; a[a_n-1].off++; } } l += (uint32_t)u->a[k]; } } return cc; } ul_idx_t *gen_ul_idx_t(const ug_opt_t *uopt, asg_t *sg, uint64_t is_el, uint64_t is_del) { uint64_t n_read = R_INF.total_reads; ma_hit_t_alloc* src = uopt->sources; int64_t min_ovlp = uopt->min_ovlp; int64_t max_hang = uopt->max_hang; // int64_t gap_fuzz = uopt->gap_fuzz; ul_idx_t *uu = NULL; CALLOC(uu, 1); uu->ug = ma_ug_gen(sg); uu->ct = ul_contain_gen(uu->ug, sg, src, min_ovlp, max_hang, is_el, is_del); uu->cc = gen_cov_track(uu->ug, sg, uu->ct, src, min_ovlp, max_hang, is_el, is_del); uu->cr = gen_r_contain(uu->ug, sg, src, n_read, min_ovlp, max_hang, asm_opt.thread_num, is_el, is_del); uu->r_ug = gen_r_ug_idx(uu->ug, sg); return uu; } void destroy_ul_idx_t(ul_idx_t *uu) { if(!uu) return; if(uu->cc) { if(uu->cc) { free(uu->cc->idx); free(uu->cc->interval.a); free(uu->cc); } if(uu->cr) { free(uu->cr->idx); free(uu->cr->interval.a); free(uu->cr); } if(uu->ct) { free(uu->ct->idx.a); free(uu->ct->rids.a); free(uu->ct->is_c.a); free(uu->ct); } if(uu->r_ug) { free(uu->r_ug->idx); free(uu->r_ug->p.a); free(uu->r_ug); } // if(uu->ov) { // free(uu->ov->a); // free(uu->ov); // } } ma_ug_destroy(uu->ug); // if(uu->nug) { // free(uu->nug->idx); // ma_ug_destroy(uu->nug->ug); // free(uu->nug); // } free(uu); } void gen_UL_ovlps(uldat_t *sl, int32_t cutoff) { ul_idx_t *uu = dedup_HiFis(sl->uopt, 1, 0); int exist = (asm_opt.load_index_from_disk? uidx_load(&ha_flt_tab, &ha_idx, asm_opt.output_file_name) : 0); if(exist == 0) uidx_l_build(uu->ug, (mg_idxopt_t *)sl->opt, cutoff); if(exist == 0) uidx_write(ha_flt_tab, ha_idx, asm_opt.output_file_name); sl->ha_flt_tab = ha_flt_tab; sl->ha_idx = (ha_pt_t *)ha_idx; sl->uu = uu; ul_v_call(sl, asm_opt.ar); destroy_ul_idx_t(uu); ha_ft_destroy(ha_flt_tab); ha_pt_destroy(ha_idx); sl->ha_flt_tab = NULL; sl->ha_idx = NULL; sl->uu = NULL; } void init_uldat_t(uldat_t *sl, void *ha_flt_tab, void *ha_idx, mg_idxopt_t *opt, uint64_t chunk_size, uint64_t n_thread, const ug_opt_t *uopt, ul_idx_t *uu) { memset(sl, 0, sizeof(uldat_t)); sl->ha_flt_tab = ha_flt_tab; sl->ha_idx = (ha_pt_t *)ha_idx; sl->opt = opt; sl->chunk_size = chunk_size; sl->n_thread = n_thread; sl->uu = uu; sl->uopt = uopt; } int32_t write_all_ul_t(all_ul_t *x, char* file_name) { char* gfa_name = NULL; MALLOC(gfa_name, strlen(file_name)+50); sprintf(gfa_name, "%s.ul.ovlp.bin", file_name); FILE* fp = fopen(gfa_name, "w"); free(gfa_name); if (!fp) return 0; uint64_t k; ul_vec_t *p = NULL; fwrite(&x->nid.n, sizeof(x->nid.n), 1, fp); for (k = 0; k < x->nid.n; k++) { fwrite(&x->nid.a[k].n, sizeof(x->nid.a[k].n), 1, fp); fwrite(x->nid.a[k].a, sizeof((*(x->nid.a[k].a))), x->nid.a[k].n, fp); } fwrite(&x->ridx.idx.n, sizeof(x->ridx.idx.n), 1, fp); fwrite(x->ridx.idx.a, sizeof((*(x->ridx.idx.a))), x->ridx.idx.n, fp); fwrite(&x->ridx.occ.n, sizeof(x->ridx.occ.n), 1, fp); fwrite(x->ridx.occ.a, sizeof((*(x->ridx.occ.a))), x->ridx.occ.n, fp); fwrite(&x->n, sizeof(x->n), 1, fp); for (k = 0; k < x->n; k++) { p = &(x->a[k]); fwrite(&p->dd, sizeof(p->dd), 1, fp); fwrite(&p->rlen, sizeof(p->rlen), 1, fp); fwrite(&p->r_base.n, sizeof(p->r_base.n), 1, fp); fwrite(p->r_base.a, sizeof((*(p->r_base.a))), p->r_base.n, fp); fwrite(&p->bb.n, sizeof(p->bb.n), 1, fp); fwrite(p->bb.a, sizeof((*(p->bb.a))), p->bb.n, fp); fwrite(&p->N_site.n, sizeof(p->N_site.n), 1, fp); fwrite(p->N_site.a, sizeof((*(p->N_site.a))), p->N_site.n, fp); } fprintf(stderr, "[M::%s] Index has been written.\n", __func__); fclose(fp); return 1; } int32_t load_all_ul_t(all_ul_t *x, char* file_name, All_reads *hR) { char* gfa_name = NULL; MALLOC(gfa_name, strlen(file_name)+50); sprintf(gfa_name, "%s.ul.ovlp.bin", file_name); FILE* fp = fopen(gfa_name, "r"); free(gfa_name); if (!fp) return 0; memset(x, 0, sizeof(*x)); x->hR = hR; init_aux_table(); uint64_t k; ul_vec_t *p = NULL; fread(&x->nid.n, sizeof(x->nid.n), 1, fp); x->nid.m = x->nid.n; MALLOC(x->nid.a, x->nid.n); for (k = 0; k < x->nid.n; k++) { fread(&x->nid.a[k].n, sizeof(x->nid.a[k].n), 1, fp); MALLOC(x->nid.a[k].a, x->nid.a[k].n); fread(x->nid.a[k].a, sizeof((*(x->nid.a[k].a))), x->nid.a[k].n, fp); } fread(&x->ridx.idx.n, sizeof(x->ridx.idx.n), 1, fp); x->ridx.idx.m = x->ridx.idx.n; MALLOC(x->ridx.idx.a, x->ridx.idx.n); fread(x->ridx.idx.a, sizeof((*(x->ridx.idx.a))), x->ridx.idx.n, fp); fread(&x->ridx.occ.n, sizeof(x->ridx.occ.n), 1, fp); x->ridx.occ.m = x->ridx.occ.n; MALLOC(x->ridx.occ.a, x->ridx.occ.n); fread(x->ridx.occ.a, sizeof((*(x->ridx.occ.a))), x->ridx.occ.n, fp); fread(&x->n, sizeof(x->n), 1, fp); x->m = x->n; MALLOC(x->a, x->n); for (k = 0; k < x->n; k++) { p = &(x->a[k]); fread(&p->dd, sizeof(p->dd), 1, fp); fread(&p->rlen, sizeof(p->rlen), 1, fp); fread(&p->r_base.n, sizeof(p->r_base.n), 1, fp); p->r_base.m = p->r_base.n; MALLOC(p->r_base.a, p->r_base.n); fread(p->r_base.a, sizeof((*(p->r_base.a))), p->r_base.n, fp); fread(&p->bb.n, sizeof(p->bb.n), 1, fp); p->bb.m = p->bb.n; MALLOC(p->bb.a, p->bb.n); fread(p->bb.a, sizeof((*(p->bb.a))), p->bb.n, fp); fread(&p->N_site.n, sizeof(p->N_site.n), 1, fp); p->N_site.m = p->N_site.n; MALLOC(p->N_site.a, p->N_site.n); fread(p->N_site.a, sizeof((*(p->N_site.a))), p->N_site.n, fp); } fprintf(stderr, "[M::%s] Index has been loaded.\n", __func__); fclose(fp); return 1; } void ul_load(const ug_opt_t *uopt) { fprintf(stderr, "[M::%s::] ==> UL\n", __func__); mg_idxopt_t opt; uldat_t sl; int32_t cutoff; init_aux_table(); ha_opt_update_cov(&asm_opt, asm_opt.hom_cov); cutoff = asm_opt.max_n_chain; init_mg_opt(&opt, !(asm_opt.flag&HA_F_NO_HPC), 19, 10, cutoff, asm_opt.max_n_chain, asm_opt.ul_error_rate, asm_opt.ul_error_rate); init_uldat_t(&sl, NULL, NULL, &opt, 500000000, asm_opt.thread_num, uopt, NULL); if(!load_all_ul_t(&UL_INF, asm_opt.output_file_name, &R_INF)) { gen_UL_ovlps(&sl, cutoff); write_all_ul_t(&UL_INF, asm_opt.output_file_name); } // print_all_ul_t_stat(&UL_INF); kt_for(sl.n_thread, update_ovlp_src, &sl, R_INF.total_reads); kt_for(sl.n_thread, update_ovlp_src_bl, &sl, R_INF.total_reads); print_ovlp_src_bl_stat(&UL_INF, sl.uopt); // print_ul_ovlps(&UL_INF, 0); print_ul_ovlps(&UL_INF, 1); // destory_all_ul_t(&UL_INF); } void ul_refine_alignment(const ug_opt_t *uopt, asg_t *sg) { fprintf(stderr, "[M::%s::] ==> UL refinement...\n", __func__); mg_idxopt_t opt; uldat_t sl; int32_t cutoff; init_aux_table(); ha_opt_update_cov(&asm_opt, asm_opt.hom_cov); cutoff = asm_opt.max_n_chain; init_mg_opt(&opt, !(asm_opt.flag&HA_F_NO_HPC), 19, 10, cutoff, asm_opt.max_n_chain, asm_opt.ul_error_rate, asm_opt.ul_error_rate); ul_idx_t *uu = gen_ul_idx_t(uopt, sg, 0, 0);///record contained reads; is_el = is_del = 0 init_uldat_t(&sl, NULL, NULL, &opt, 500000000, asm_opt.thread_num, uopt, uu); sl.rg = sg; work_ul_gchains(&sl); destroy_ul_idx_t(uu); }