diff --git a/align.c b/align.c index 48b4478..9035b42 100644 --- a/align.c +++ b/align.c @@ -300,9 +300,8 @@ static void mm_update_extra(mm_reg1_t *r, const uint8_t *qseq, const uint8_t *ts if (is_eqx) mm_update_cigar_eqx(r, qseq, tseq); // NB: it has to be called here as changes to qseq and tseq are not returned } -static void mm_append_cigar(mm_reg1_t *r, uint32_t n_cigar, uint32_t *cigar) // TODO: this calls the libc realloc() +void mm_enlarge_cigar(mm_reg1_t *r, uint32_t n_cigar) // TODO: this calls the libc realloc() { - mm_extra_t *p; if (n_cigar == 0) return; if (r->p == 0) { uint32_t capacity = n_cigar + sizeof(mm_extra_t)/4; @@ -314,6 +313,13 @@ static void mm_append_cigar(mm_reg1_t *r, uint32_t n_cigar, uint32_t *cigar) // kroundup32(r->p->capacity); r->p = (mm_extra_t*)realloc(r->p, r->p->capacity * 4); } +} + +static void mm_append_cigar(mm_reg1_t *r, uint32_t n_cigar, const uint32_t *cigar) +{ + mm_extra_t *p; + if (n_cigar == 0) return; + mm_enlarge_cigar(r, n_cigar); p = r->p; if (p->n_cigar > 0 && (p->cigar[p->n_cigar-1]&0xf) == (cigar[0]&0xf)) { // same CIGAR op at the boundary p->cigar[p->n_cigar-1] += cigar[0]>>4<<4; diff --git a/hit.c b/hit.c index f5a50a2..ecfae91 100644 --- a/hit.c +++ b/hit.c @@ -464,3 +464,170 @@ void mm_set_mapq(void *km, int n_regs, mm_reg1_t *regs, int min_chain_sc, int ma } mm_set_inv_mapq(km, n_regs, regs); } + +/****************************** + * Instructed split alignment * + ******************************/ + +#define MM_MIN_EXON_LEN 20 + +static int32_t mm_jump_check(void *km, const mm_idx_t *mi, int32_t qlen, const uint8_t *qseq0, const mm_reg1_t *r, int32_t ext, int32_t is_left) // TODO: check close N +{ + int32_t clip, clen, e = !r->rev ^ !is_left; // 0 for left of the alignment; 1 for right + uint32_t cigar; + if (!r->p || r->p->n_cigar <= 0) return -1; // only working with CIGAR + clip = e == 0? r->qs : qlen - r->qe; + cigar = r->p->cigar[e == 0? 0 : r->p->n_cigar - 1]; + clen = (cigar&0xf) == MM_CIGAR_MATCH? cigar>>4 : 0; + if (clen <= ext) return -1; + if (is_left) { + if (clip >= r->rs) return -1; // no space to jump + } else { + if (clip >= mi->seq[r->rid].len - r->re) return -1; // no space to jump + } + return 0; +} + +static uint8_t *mm_jump_get_qseq_seq(void *km, int32_t qlen, const uint8_t *qseq0, const mm_reg1_t *r, int32_t is_left, int32_t ql0, uint8_t *qseq) +{ + int32_t i, k; + if (!r->rev) { + if (is_left) memcpy(qseq, qseq0, ql0); + else memcpy(qseq, &qseq0[qlen - ql0], ql0); + } else { + if (is_left) + for (i = qlen - 1, k = 0; i >= qlen - ql0; --i) + qseq[k++] = qseq0[i] >= 4? qseq0[i] : 3 - qseq0[i]; + else + for (i = ql0 - 1, k = 0; i >= 0; --i) + qseq[k++] = qseq0[i] >= 4? qseq0[i] : 3 - qseq0[i]; + } + return qseq; +} + +static void mm_jump_split_left(void *km, const mm_idx_t *mi, const mm_mapopt_t *opt, int32_t qlen, const uint8_t *qseq0, mm_reg1_t *r, int32_t ts_strand) +{ + uint8_t *tseq = 0, *qseq = 0; + int32_t i, n, i0 = -1, m = 0, l; + int32_t ext = 1 + (opt->b + opt->a - 1) / opt->a + 1; + int32_t clip = !r->rev? r->qs : qlen - r->qe; + int32_t extt = clip < ext? clip : ext; + const mm_idx_jjump1_t *a; + + if (mm_jump_check(km, mi, qlen, qseq0, r, ext + MM_MIN_EXON_LEN, 1) < 0) return; + a = mm_idx_jump_get(mi, r->rid, r->rs - extt, r->rs + ext, &n); + if (n == 0) return; + + for (i = 0; i < n; ++i) { // traverse possible jumps + const mm_idx_jjump1_t *ai = &a[i]; + int32_t tlen, tl1, j, mm1, mm2; + assert(ai->off >= r->rs - extt && ai->off < r->rs + ext); + if (ts_strand * ai->strand < 0) continue; // wrong strand + if (ai->off2 >= ai->off) continue; // wrong direction + if (ai->off2 < clip + ext) continue; // not long enough + if (tseq == 0) { + tseq = Kcalloc(km, uint8_t, (clip + ext) * 2); // tseq and qseq are allocated together + qseq = tseq + clip + ext; + mm_jump_get_qseq_seq(km, qlen, qseq0, r, 1, clip + ext, qseq); + } + tl1 = clip + (ai->off - r->rs); + tlen = mm_idx_getseq2(mi, 0, r->rid, ai->off, r->rs + ext, &tseq[tl1]); + assert(tlen == r->rs + ext - ai->off); + tlen = mm_idx_getseq2(mi, 0, r->rid, ai->off2 - tl1, ai->off2, tseq); + assert(tlen == tl1); + for (j = 0, mm1 = 0; j < tl1; ++j) + if (qseq[j] != tseq[j] || qseq[j] > 3 || tseq[j] > 3) + ++mm1; + for (mm2 = 0; j < clip + ext; ++j) + if (qseq[j] != tseq[j] || qseq[j] > 3 || tseq[j] > 3) + ++mm2; + if (mm1 == 0 && mm2 == 1) + i0 = i, ++m; // i0 points to the rightmost i + } + kfree(km, tseq); + + l = m > 0? a[i0].off - r->rs : 0; // may be negative + if (m == 1 && clip + l >= opt->jump_min_alen) { // add one more exon + mm_enlarge_cigar(r, 2); + memmove(r->p->cigar + 2, r->p->cigar, r->p->n_cigar * 4); + r->p->cigar[0] = (clip + l) << 4 | MM_CIGAR_MATCH; + r->p->cigar[1] = (a[i0].off - a[i0].off2) << 4 | MM_CIGAR_N_SKIP; + r->p->cigar[2] = ((r->p->cigar[2]>>4) - l) << 4 | MM_CIGAR_MATCH; + r->p->n_cigar += 2; + r->rs = a[i0].off2 - (clip + l); + if (!r->rev) r->qs = 0; + else r->qe = qlen; + } else if (m > 0 && a[i0].off > r->rs) { // trim by l; l is always positive + r->p->cigar[0] -= l << 4 | MM_CIGAR_MATCH; + r->rs += l; + if (!r->rev) r->qs += l; + else r->qe -= l; + } +} + +static void mm_jump_split_right(void *km, const mm_idx_t *mi, const mm_mapopt_t *opt, int32_t qlen, const uint8_t *qseq0, mm_reg1_t *r, int32_t ts_strand) +{ + uint8_t *tseq = 0, *qseq = 0; + int32_t i, n, i0 = -1, m = 0, l; + int32_t ext = 1 + (opt->b + opt->a - 1) / opt->a + 1; + int32_t clip = !r->rev? qlen - r->qe : r->qs; + int32_t extt = clip < ext? clip : ext; + const mm_idx_jjump1_t *a; + + if (mm_jump_check(km, mi, qlen, qseq0, r, ext + MM_MIN_EXON_LEN, 1) < 0) return; + a = mm_idx_jump_get(mi, r->rid, r->re - ext, r->re + extt, &n); + if (n == 0) return; + + for (i = 0; i < n; ++i) { // traverse possible jumps + const mm_idx_jjump1_t *ai = &a[i]; + int32_t tlen, tl1, j, mm1, mm2; + assert(ai->off >= r->rs - extt && ai->off < r->rs + ext); + if (ts_strand * ai->strand < 0) continue; // wrong strand + if (ai->off2 <= ai->off) continue; // wrong direction + if (ai->off2 + clip + ext > mi->seq[r->rid].len) continue; // not long enough + if (tseq == 0) { + tseq = Kcalloc(km, uint8_t, (clip + ext) * 2); // tseq and qseq are allocated together + qseq = tseq + clip + ext; + mm_jump_get_qseq_seq(km, qlen, qseq0, r, 0, clip + ext, qseq); + } + tl1 = clip + (r->re - ai->off); + tlen = mm_idx_getseq2(mi, 0, r->rid, r->re - ext, ai->off, tseq); + assert(tlen == ai->off - (r->re - ext)); + tlen = mm_idx_getseq2(mi, 0, r->rid, ai->off2, ai->off2 + tl1, &tseq[clip + ext - tl1]); + assert(tlen == tl1); + for (j = 0, mm2 = 0; j < clip + ext - tl1; ++j) + if (qseq[j] != tseq[j] || qseq[j] > 3 || tseq[j] > 3) + ++mm2; + for (mm1 = 0; j < clip + ext; ++j) + if (qseq[j] != tseq[j] || qseq[j] > 3 || tseq[j] > 3) + ++mm1; + if (mm1 == 0 && mm2 == 1) + i0 = i0 >= 0? i0 : i, ++m; // i0 points to the leftmost i + } + kfree(km, tseq); + + l = m > 0? r->re - a[i0].off : 0; // may be negative + if (m == 1 && clip + l >= opt->jump_min_alen) { // add one more exon + mm_enlarge_cigar(r, 2); + memmove(r->p->cigar + 2, r->p->cigar, r->p->n_cigar * 4); + r->p->cigar[r->p->n_cigar - 1] = ((r->p->cigar[r->p->n_cigar - 1]>>4) - l) << 4 | MM_CIGAR_MATCH; + r->p->cigar[r->p->n_cigar] = (a[i0].off2 - a[i0].off) << 4 | MM_CIGAR_N_SKIP; + r->p->cigar[r->p->n_cigar + 1] = (clip + l) << 4 | MM_CIGAR_MATCH; + r->p->n_cigar += 2; + r->re = a[i0].off2 + (clip + l); + if (!r->rev) r->qe = qlen; + else r->qs = 0; + } else if (m > 0 && r->re > a[i0].off) { // trim by l; l is always positive + r->p->cigar[r->p->n_cigar - 1] -= l << 4 | MM_CIGAR_MATCH; + r->re -= l; + if (!r->rev) r->qe -= l; + else r->qs += l; + } +} + +void mm_jump_split(void *km, const mm_idx_t *mi, const mm_mapopt_t *opt, int32_t qlen, const uint8_t *qseq, mm_reg1_t *r, int32_t ts_strand) +{ + assert((opt->flag & MM_F_EQX) == 0); + mm_jump_split_left(km, mi, opt, qlen, qseq, r, ts_strand); + mm_jump_split_right(km, mi, opt, qlen, qseq, r, ts_strand); +} diff --git a/index.c b/index.c index 30ad28b..54b89a4 100644 --- a/index.c +++ b/index.c @@ -855,6 +855,35 @@ int mm_idx_bed_junc(const mm_idx_t *mi, int32_t ctg, int32_t st, int32_t en, uin return left; } +static int32_t mm_idx_jump_get_core(int32_t n, const mm_idx_jjump1_t *a, int32_t x) // similar to mm_idx_find_intv() +{ + int32_t s = 0, e = n; + if (n == 0) return -1; + if (x < a[0].off) return -1; + while (s < e) { + int32_t mid = s + (e - s) / 2; + if (x >= a[mid].off && (mid + 1 >= n || x < a[mid+1].off)) return mid; + else if (x < a[mid].off) e = mid; + else s = mid + 1; + } + assert(0); +} + +const mm_idx_jjump1_t *mm_idx_jump_get(const mm_idx_t *db, int32_t cid, int32_t st, int32_t en, int32_t *n) +{ + mm_idx_jjump_t *s; + int32_t l, r; + *n = 0; + if (cid >= db->n_seq || cid < 0 || db->J == 0) return 0; + if (en < 0 || en > db->seq[cid].len) en = db->seq[cid].len; + s = &db->J[cid]; + if (s->n == 0) return 0; + l = mm_idx_jump_get_core(s->n, s->a, st); + r = mm_idx_jump_get_core(s->n, s->a, en); + *n = r - l; + return &s->a[l + 1]; +} + /**************** * splice score * ****************/ diff --git a/minimap.h b/minimap.h index 7742272..cb2ff32 100644 --- a/minimap.h +++ b/minimap.h @@ -174,6 +174,8 @@ typedef struct { int pe_ori, pe_bonus; + int32_t jump_min_alen; + float mid_occ_frac; // only used by mm_mapopt_update(); see below float q_occ_frac; int32_t min_mid_occ, max_mid_occ; diff --git a/mmpriv.h b/mmpriv.h index 83f9ee7..bc71d7b 100644 --- a/mmpriv.h +++ b/mmpriv.h @@ -83,8 +83,8 @@ void mm_idxopt_init(mm_idxopt_t *opt); const uint64_t *mm_idx_get(const mm_idx_t *mi, uint64_t minier, int *n); int32_t mm_idx_cal_max_occ(const mm_idx_t *mi, float f); int mm_idx_getseq2(const mm_idx_t *mi, int is_rev, uint32_t rid, uint32_t st, uint32_t en, uint8_t *seq); -mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, int *n_regs_, mm_reg1_t *regs, mm128_t *a); mm_reg1_t *mm_gen_regs(void *km, uint32_t hash, int qlen, int n_u, uint64_t *u, mm128_t *a, int is_qstrand); +const mm_idx_jjump1_t *mm_idx_jump_get(const mm_idx_t *db, int32_t cid, int32_t st, int32_t en, int32_t *n); mm128_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, float chn_pen_skip, int is_cdna, int n_segs, int64_t n, mm128_t *a, int *n_u_, uint64_t **_u, void *km); @@ -104,6 +104,10 @@ void mm_filter_regs(const mm_mapopt_t *opt, int qlen, int *n_regs, mm_reg1_t *re void mm_hit_sort(void *km, int *n_regs, mm_reg1_t *r, float alt_diff_frac); void mm_set_mapq(void *km, int n_regs, mm_reg1_t *regs, int min_chain_sc, int match_sc, int rep_len, int is_sr); void mm_update_dp_max(int qlen, int n_regs, mm_reg1_t *regs, float frac, int a, int b); +void mm_jump_split(void *km, const mm_idx_t *mi, const mm_mapopt_t *opt, int32_t qlen, const uint8_t *qseq, mm_reg1_t *r, int32_t ts_strand); + +mm_reg1_t *mm_align_skeleton(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, const char *qstr, int *n_regs_, mm_reg1_t *regs, mm128_t *a); +void mm_enlarge_cigar(mm_reg1_t *r, uint32_t n_cigar); void mm_est_err(const mm_idx_t *mi, int qlen, int n_regs, mm_reg1_t *regs, const mm128_t *a, int32_t n, const uint64_t *mini_pos); diff --git a/options.c b/options.c index 547e5bc..22927db 100644 --- a/options.c +++ b/options.c @@ -62,6 +62,8 @@ void mm_mapopt_init(mm_mapopt_t *opt) opt->pe_ori = 0; // FF opt->pe_bonus = 33; + + opt->jump_min_alen = 10; } void mm_mapopt_update(mm_mapopt_t *opt, const mm_idx_t *mi)