#include #include #include "minimap.h" #include "ksw2.h" #ifndef kroundup32 #define kroundup32(x) (--(x), (x)|=(x)>>1, (x)|=(x)>>2, (x)|=(x)>>4, (x)|=(x)>>8, (x)|=(x)>>16, ++(x)) #endif static void ksw_gen_simple_mat(int m, int8_t *mat, int8_t a, int8_t b) { int i, j; a = a < 0? -a : a; b = b > 0? -b : b; for (i = 0; i < m - 1; ++i) { for (j = 0; j < m - 1; ++j) mat[i * m + j] = i == j? a : b; mat[i * m + m - 1] = 0; } for (j = 0; j < m; ++j) mat[(m - 1) * m + j] = 0; } static inline void mm_seq_rev(uint32_t len, uint8_t *seq) { uint32_t i; uint8_t t; for (i = 0; i < len>>1; ++i) t = seq[i], seq[i] = seq[len - 1 - i], seq[len - 1 - i] = t; } static void mm_update_extra(mm_extra_t *p, const uint8_t *qseq, const uint8_t *tseq, uint32_t n_cigar, uint32_t *cigar) { uint32_t k, l, toff = 0, qoff = 0; for (k = 0; k < n_cigar; ++k) { uint32_t op = cigar[k]&0xf, len = cigar[k]>>4; if (op == 0) { for (l = 0; l < len; ++l) { if (tseq[toff + l] > 3 || qseq[qoff + l] > 3) ++p->n_ambi; else if (tseq[toff + l] != qseq[qoff + l]) ++p->n_diff; } toff += len, qoff += len, p->blen += len; } else if (op == 1) { int n_ambi = 0; for (l = 0; l < len; ++l) if (qseq[qoff + l] > 3) ++n_ambi; qoff += len, p->blen += len; p->n_ambi += n_ambi, p->n_diff += len - n_ambi; } else if (op == 2) { int n_ambi = 0; for (l = 0; l < len; ++l) if (tseq[toff + l] > 3) ++n_ambi; toff += len, p->blen += len; p->n_ambi += n_ambi, p->n_diff += len - n_ambi; } } } static void mm_append_cigar(mm_reg1_t *r, uint32_t n_cigar, uint32_t *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); kroundup32(capacity); r->p = (mm_extra_t*)calloc(capacity, 4); r->p->capacity = capacity; } else if (r->p->n_cigar + n_cigar + sizeof(mm_extra_t) > r->p->capacity) { r->p->capacity = r->p->n_cigar + n_cigar + sizeof(mm_extra_t); kroundup32(r->p->capacity); r->p = (mm_extra_t*)realloc(r->p, r->p->capacity * 4); } 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; if (n_cigar > 1) memcpy(p->cigar + p->n_cigar, cigar + 1, (n_cigar - 1) * 4); p->n_cigar += n_cigar - 1; } else { memcpy(p->cigar + p->n_cigar, cigar, n_cigar * 4); p->n_cigar += n_cigar; } } static inline void mm_adjust_minier(const mm_idx_t *mi, uint8_t *qseq0[2], uint8_t *tseq0, mm128_t *a, int32_t *r, int32_t *q) { if (mi->is_hpc && 1) { int span, rev, i, r0, c; rev = a->x >> 63; *q = (int32_t)a->y; for (i = *q - 1, c = qseq0[rev][*q]; i > 0; --i) if (qseq0[rev][i] != c) break; *q = i + 1; span = a->y >> 32; *r = (int32_t)a->x; r0 = *r + 1 >= 256? *r + 1 - 256 : 0; mm_idx_getseq(mi, a->x<<1>>33, r0, *r + 1, tseq0); for (i = *r - 1 - r0, c = tseq0[*r - r0]; i > 0; --i) if (tseq0[i] != c) break; *r = i + r0 + 1; } else { *r = (int32_t)a->x + 1; *q = (int32_t)a->y + 1; } } static void mm_align1(void *km, const mm_mapopt_t *opt, const mm_idx_t *mi, int qlen, uint8_t *qseq0[2], mm_reg1_t *r, mm_reg1_t *r2, mm128_t *a, ksw_extz_t *ez) { int32_t rid = a[r->as].x<<1>>33, rev = a[r->as].x>>63; uint8_t *tseq, *qseq; int32_t i, l, bw, rs0, re0, qs0, qe0; int32_t rs, re, qs, qe; int32_t rs1, qs1, re1, qe1; int8_t mat[25]; ksw_gen_simple_mat(5, mat, opt->a, opt->b); bw = (int)(opt->bw * 1.5 + 1.); /* rs = (int32_t)a[r->as].x + 1; // NB: this is the same as r->{rs,re} re = (int32_t)a[r->as + r->cnt - 1].x + 1; qs = (int32_t)a[r->as].y + 1; // NB: this is the coordinate on the reverse strand; r->{qs,qe} are on the reverse strand qe = (int32_t)a[r->as + r->cnt - 1].y + 1; */ tseq = (uint8_t*)kmalloc(km, 256); mm_adjust_minier(mi, qseq0, tseq, &a[r->as], &rs, &qs); mm_adjust_minier(mi, qseq0, tseq, &a[r->as + r->cnt - 1], &re, &qe); kfree(km, tseq); if (qs > 0 && rs > 0) { l = qs < opt->max_gap? qs : opt->max_gap; qs0 = qs - l; l += (l * opt->a - opt->q) / opt->e; l = l < opt->max_gap? l : opt->max_gap; l = l < rs? l : rs; rs0 = rs - l; } else rs0 = rs, qs0 = qs; if (qe < qlen && re < mi->seq[rid].len) { l = qlen - qe < opt->max_gap? qlen - qe : opt->max_gap; qe0 = qe + l; l += (l * opt->a - opt->q) / opt->e; l = l < opt->max_gap? l : opt->max_gap; l = l < mi->seq[rid].len - re? l : mi->seq[rid].len - re; re0 = re + l; } else re0 = re, qe0 = qe; tseq = (uint8_t*)kmalloc(km, re0 - rs0 > 256? re0 - rs0 : 256); // TODO: we can allocate a smaller size if (qs > 0 && rs > 0) { // left extension uint32_t ql = qs - qs0, tl = rs - rs0; qseq = &qseq0[rev][qs0]; mm_idx_getseq(mi, rid, rs0, rs, tseq); mm_seq_rev(ql, qseq); mm_seq_rev(tl, tseq); #if 0 fprintf(stderr, "===> [-1] %d-%d %c (%s:%d-%d) <===\n", qs0, qs, "+-"[rev], mi->seq[rid].name, rs0, rs); for (k = 0; k < tl; ++k) fputc("ACGTN"[tseq[k]], stderr); fputc('\n', stderr); for (k = 0; k < ql; ++k) fputc("ACGTN"[qseq[k]], stderr); fputc('\n', stderr); #endif ksw_extz2_sse(km, ql, qseq, tl, tseq, 5, mat, opt->q, opt->e, bw, opt->zdrop, KSW_EZ_EXTZ_ONLY|KSW_EZ_RIGHT|KSW_EZ_REV_CIGAR, ez); mm_append_cigar(r, ez->n_cigar, ez->cigar); mm_update_extra(r->p, qseq, tseq, ez->n_cigar, ez->cigar); r->p->score += ez->max; rs1 = rs - (ez->max_t + 1); qs1 = qs - (ez->max_q + 1); mm_seq_rev(ql, qseq); } else rs1 = rs, qs1 = qs; assert(qs1 >= 0 && rs1 >= 0); for (i = 1; i < r->cnt; ++i) { // gap filling mm_adjust_minier(mi, qseq0, tseq, &a[r->as + i], &re, &qe); if (i == r->cnt - 1 || qe - qs >= opt->min_ksw_len || re - rs >= opt->min_ksw_len) { qseq = &qseq0[rev][qs]; mm_idx_getseq(mi, rid, rs, re, tseq); #if 0 fprintf(stderr, "===> [%d] %d-%d %c (%s:%d-%d) <===\n", i, qs, qe, "+-"[rev], mi->seq[rid].name, rs, re); for (k = 0; k < re - rs; ++k) fputc("ACGTN"[tseq[k]], stderr); fputc('\n', stderr); for (k = 0; k < qe - qs; ++k) fputc("ACGTN"[qseq[k]], stderr); fputc('\n', stderr); #endif ksw_extz2_sse(km, qe-qs, qseq, re-rs, tseq, 5, mat, opt->q, opt->e, bw, opt->zdrop, KSW_EZ_DYN_BAND, ez); mm_append_cigar(r, ez->n_cigar, ez->cigar); mm_update_extra(r->p, qseq, tseq, ez->n_cigar, ez->cigar); if (ez->score == KSW_NEG_INF) { // truncated by Z-drop r->p->score += ez->max; abort(); } else { r->p->score += ez->score; } //for (k = 0; k < r->cigar->n; ++k) fprintf(stderr, "%d%c", r->cigar->cigar[k]>>4, "MID"[r->cigar->cigar[k]&0xf]); fputc('\n', stderr); rs = re, qs = qe; } } if (i == r->cnt && qe < qe0 && re < re0) { // right extension qseq = &qseq0[rev][qe]; mm_idx_getseq(mi, rid, re, re0, tseq); ksw_extz2_sse(km, qe0-qe, qseq, re0-re, tseq, 5, mat, opt->q, opt->e, bw, opt->zdrop, KSW_EZ_EXTZ_ONLY, ez); mm_append_cigar(r, ez->n_cigar, ez->cigar); mm_update_extra(r->p, qseq, tseq, ez->n_cigar, ez->cigar); r->p->score += ez->max; re1 = re + (ez->max_t + 1); qe1 = qe + (ez->max_q + 1); } else re1 = re, qe1 = qe; assert(qe1 <= qlen); r->rs = rs1, r->re = re1; if (rev) r->qs = qlen - qe1, r->qe = qlen - qs1; else r->qs = qs1, r->qe = qe1; // for (i = 0; i < r->p->n_cigar; ++i) fprintf(stderr, "%d%c", r->p->cigar[i]>>4, "MID"[r->p->cigar[i]&0xf]); fputc('\n', stderr); kfree(km, tseq); } void 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) { extern unsigned char seq_nt4_table[256]; int i, reg; uint8_t *qseq0[2]; ksw_extz_t ez; memset(&ez, 0, sizeof(ksw_extz_t)); qseq0[0] = (uint8_t*)kmalloc(km, qlen); qseq0[1] = (uint8_t*)kmalloc(km, qlen); for (i = 0; i < qlen; ++i) { qseq0[0][i] = seq_nt4_table[(uint8_t)qstr[i]]; qseq0[1][qlen - 1 - i] = qseq0[0][i] < 4? 3 - qseq0[0][i] : 4; } for (reg = 0; reg < n_regs; ++reg) { mm_reg1_t r2; mm_align1(km, opt, mi, qlen, qseq0, ®s[reg], &r2, a, &ez); } kfree(km, qseq0[0]); kfree(km, qseq0[1]); kfree(km, ez.cigar); }