#include #include #include #include #include "bam.h" #include "bam_endian.h" #include "kstring.h" #include "sam_header.h" int bam_is_be = 0, bam_verbose = 2, bam_no_B = 0; char *bam_flag2char_table = "pPuUrR12sfd\0\0\0\0\0"; /************************** * CIGAR related routines * **************************/ uint32_t bam_calend(const bam1_core_t *c, const uint32_t *cigar) { int k, end = c->pos; for (k = 0; k < c->n_cigar; ++k) { int op = bam_cigar_op(cigar[k]); int len = bam_cigar_oplen(cigar[k]); if (op == BAM_CBACK) { // move backward int l, u, v; if (k == c->n_cigar - 1) break; // skip trailing 'B' for (l = k - 1, u = v = 0; l >= 0; --l) { int op1 = bam_cigar_op(cigar[l]); int len1 = bam_cigar_oplen(cigar[l]); if (bam_cigar_type(op1)&1) { // consume query if (u + len1 >= len) { // stop if (bam_cigar_type(op1)&2) v += len - u; break; } else u += len1; } if (bam_cigar_type(op1)&2) v += len1; } end = l < 0? c->pos : end - v; } else if (bam_cigar_type(op)&2) end += bam_cigar_oplen(cigar[k]); } return end; } int32_t bam_cigar2qlen(const bam1_core_t *c, const uint32_t *cigar) { uint32_t k; int32_t l = 0; for (k = 0; k < c->n_cigar; ++k) if (bam_cigar_type(bam_cigar_op(cigar[k]))&1) l += bam_cigar_oplen(cigar[k]); return l; } /******************** * BAM I/O routines * ********************/ bam_header_t *bam_header_init() { bam_is_be = bam_is_big_endian(); return (bam_header_t*)calloc(1, sizeof(bam_header_t)); } void bam_header_destroy(bam_header_t *header) { int32_t i; extern void bam_destroy_header_hash(bam_header_t *header); if (header == 0) return; if (header->target_name) { for (i = 0; i < header->n_targets; ++i) free(header->target_name[i]); free(header->target_name); free(header->target_len); } free(header->text); if (header->dict) sam_header_free(header->dict); if (header->rg2lib) sam_tbl_destroy(header->rg2lib); bam_destroy_header_hash(header); free(header); } bam_header_t *bam_header_read(bamFile fp) { bam_header_t *header; char buf[4]; int magic_len; int32_t i = 1, name_len; // check EOF i = bgzf_check_EOF(fp); if (i < 0) { // If the file is a pipe, checking the EOF marker will *always* fail // with ESPIPE. Suppress the error message in this case. if (errno != ESPIPE) perror("[bam_header_read] bgzf_check_EOF"); } else if (i == 0) fprintf(stderr, "[bam_header_read] EOF marker is absent. The input is probably truncated.\n"); // read "BAM1" magic_len = bam_read(fp, buf, 4); if (magic_len != 4 || strncmp(buf, "BAM\001", 4) != 0) { fprintf(stderr, "[bam_header_read] invalid BAM binary header (this is not a BAM file).\n"); return 0; } header = bam_header_init(); // read plain text and the number of reference sequences bam_read(fp, &header->l_text, 4); if (bam_is_be) bam_swap_endian_4p(&header->l_text); header->text = (char*)calloc(header->l_text + 1, 1); bam_read(fp, header->text, header->l_text); bam_read(fp, &header->n_targets, 4); if (bam_is_be) bam_swap_endian_4p(&header->n_targets); // read reference sequence names and lengths header->target_name = (char**)calloc(header->n_targets, sizeof(char*)); header->target_len = (uint32_t*)calloc(header->n_targets, 4); for (i = 0; i != header->n_targets; ++i) { bam_read(fp, &name_len, 4); if (bam_is_be) bam_swap_endian_4p(&name_len); header->target_name[i] = (char*)calloc(name_len, 1); bam_read(fp, header->target_name[i], name_len); bam_read(fp, &header->target_len[i], 4); if (bam_is_be) bam_swap_endian_4p(&header->target_len[i]); } return header; } int bam_header_write(bamFile fp, const bam_header_t *header) { char buf[4]; int32_t i, name_len, x; // write "BAM1" strncpy(buf, "BAM\001", 4); bam_write(fp, buf, 4); // write plain text and the number of reference sequences if (bam_is_be) { x = bam_swap_endian_4(header->l_text); bam_write(fp, &x, 4); if (header->l_text) bam_write(fp, header->text, header->l_text); x = bam_swap_endian_4(header->n_targets); bam_write(fp, &x, 4); } else { bam_write(fp, &header->l_text, 4); if (header->l_text) bam_write(fp, header->text, header->l_text); bam_write(fp, &header->n_targets, 4); } // write sequence names and lengths for (i = 0; i != header->n_targets; ++i) { char *p = header->target_name[i]; name_len = strlen(p) + 1; if (bam_is_be) { x = bam_swap_endian_4(name_len); bam_write(fp, &x, 4); } else bam_write(fp, &name_len, 4); bam_write(fp, p, name_len); if (bam_is_be) { x = bam_swap_endian_4(header->target_len[i]); bam_write(fp, &x, 4); } else bam_write(fp, &header->target_len[i], 4); } bgzf_flush(fp); return 0; } static void swap_endian_data(const bam1_core_t *c, int data_len, uint8_t *data) { uint8_t *s; uint32_t i, *cigar = (uint32_t*)(data + c->l_qname); s = data + c->n_cigar*4 + c->l_qname + c->l_qseq + (c->l_qseq + 1)/2; for (i = 0; i < c->n_cigar; ++i) bam_swap_endian_4p(&cigar[i]); while (s < data + data_len) { uint8_t type; s += 2; // skip key type = toupper(*s); ++s; // skip type if (type == 'C' || type == 'A') ++s; else if (type == 'S') { bam_swap_endian_2p(s); s += 2; } else if (type == 'I' || type == 'F') { bam_swap_endian_4p(s); s += 4; } else if (type == 'D') { bam_swap_endian_8p(s); s += 8; } else if (type == 'Z' || type == 'H') { while (*s) ++s; ++s; } else if (type == 'B') { int32_t n, Bsize = bam_aux_type2size(*s); memcpy(&n, s + 1, 4); if (1 == Bsize) { } else if (2 == Bsize) { for (i = 0; i < n; i += 2) bam_swap_endian_2p(s + 5 + i); } else if (4 == Bsize) { for (i = 0; i < n; i += 4) bam_swap_endian_4p(s + 5 + i); } bam_swap_endian_4p(s+1); } } } int bam_read1(bamFile fp, bam1_t *b) { bam1_core_t *c = &b->core; int32_t block_len, ret, i; uint32_t x[8]; assert(BAM_CORE_SIZE == 32); if ((ret = bam_read(fp, &block_len, 4)) != 4) { if (ret == 0) return -1; // normal end-of-file else return -2; // truncated } if (bam_read(fp, x, BAM_CORE_SIZE) != BAM_CORE_SIZE) return -3; if (bam_is_be) { bam_swap_endian_4p(&block_len); for (i = 0; i < 8; ++i) bam_swap_endian_4p(x + i); } c->tid = x[0]; c->pos = x[1]; c->bin = x[2]>>16; c->qual = x[2]>>8&0xff; c->l_qname = x[2]&0xff; c->flag = x[3]>>16; c->n_cigar = x[3]&0xffff; c->l_qseq = x[4]; c->mtid = x[5]; c->mpos = x[6]; c->isize = x[7]; b->data_len = block_len - BAM_CORE_SIZE; if (b->m_data < b->data_len) { b->m_data = b->data_len; kroundup32(b->m_data); b->data = (uint8_t*)realloc(b->data, b->m_data); } if (bam_read(fp, b->data, b->data_len) != b->data_len) return -4; b->l_aux = b->data_len - c->n_cigar * 4 - c->l_qname - c->l_qseq - (c->l_qseq+1)/2; if (bam_is_be) swap_endian_data(c, b->data_len, b->data); if (bam_no_B) bam_remove_B(b); return 4 + block_len; } inline int bam_write1_core(bamFile fp, const bam1_core_t *c, int data_len, uint8_t *data) { uint32_t x[8], block_len = data_len + BAM_CORE_SIZE, y; int i; assert(BAM_CORE_SIZE == 32); x[0] = c->tid; x[1] = c->pos; x[2] = (uint32_t)c->bin<<16 | c->qual<<8 | c->l_qname; x[3] = (uint32_t)c->flag<<16 | c->n_cigar; x[4] = c->l_qseq; x[5] = c->mtid; x[6] = c->mpos; x[7] = c->isize; bgzf_flush_try(fp, 4 + block_len); if (bam_is_be) { for (i = 0; i < 8; ++i) bam_swap_endian_4p(x + i); y = block_len; bam_write(fp, bam_swap_endian_4p(&y), 4); swap_endian_data(c, data_len, data); } else bam_write(fp, &block_len, 4); bam_write(fp, x, BAM_CORE_SIZE); bam_write(fp, data, data_len); if (bam_is_be) swap_endian_data(c, data_len, data); return 4 + block_len; } int bam_write1(bamFile fp, const bam1_t *b) { return bam_write1_core(fp, &b->core, b->data_len, b->data); } char *bam_format1_core(const bam_header_t *header, const bam1_t *b, int of) { uint8_t *s = bam1_seq(b), *t = bam1_qual(b); int i; const bam1_core_t *c = &b->core; kstring_t str; str.l = str.m = 0; str.s = 0; kputsn(bam1_qname(b), c->l_qname-1, &str); kputc('\t', &str); if (of == BAM_OFDEC) { kputw(c->flag, &str); kputc('\t', &str); } else if (of == BAM_OFHEX) ksprintf(&str, "0x%x\t", c->flag); else { // BAM_OFSTR for (i = 0; i < 16; ++i) if ((c->flag & 1<tid < 0) kputsn("*\t", 2, &str); else { if (header) kputs(header->target_name[c->tid] , &str); else kputw(c->tid, &str); kputc('\t', &str); } kputw(c->pos + 1, &str); kputc('\t', &str); kputw(c->qual, &str); kputc('\t', &str); if (c->n_cigar == 0) kputc('*', &str); else { uint32_t *cigar = bam1_cigar(b); for (i = 0; i < c->n_cigar; ++i) { kputw(bam1_cigar(b)[i]>>BAM_CIGAR_SHIFT, &str); kputc(bam_cigar_opchr(cigar[i]), &str); } } kputc('\t', &str); if (c->mtid < 0) kputsn("*\t", 2, &str); else if (c->mtid == c->tid) kputsn("=\t", 2, &str); else { if (header) kputs(header->target_name[c->mtid], &str); else kputw(c->mtid, &str); kputc('\t', &str); } kputw(c->mpos + 1, &str); kputc('\t', &str); kputw(c->isize, &str); kputc('\t', &str); if (c->l_qseq) { for (i = 0; i < c->l_qseq; ++i) kputc(bam_nt16_rev_table[bam1_seqi(s, i)], &str); kputc('\t', &str); if (t[0] == 0xff) kputc('*', &str); else for (i = 0; i < c->l_qseq; ++i) kputc(t[i] + 33, &str); } else kputsn("*\t*", 3, &str); s = bam1_aux(b); while (s < b->data + b->data_len) { uint8_t type, key[2]; key[0] = s[0]; key[1] = s[1]; s += 2; type = *s; ++s; kputc('\t', &str); kputsn((char*)key, 2, &str); kputc(':', &str); if (type == 'A') { kputsn("A:", 2, &str); kputc(*s, &str); ++s; } else if (type == 'C') { kputsn("i:", 2, &str); kputw(*s, &str); ++s; } else if (type == 'c') { kputsn("i:", 2, &str); kputw(*(int8_t*)s, &str); ++s; } else if (type == 'S') { kputsn("i:", 2, &str); kputw(*(uint16_t*)s, &str); s += 2; } else if (type == 's') { kputsn("i:", 2, &str); kputw(*(int16_t*)s, &str); s += 2; } else if (type == 'I') { kputsn("i:", 2, &str); kputuw(*(uint32_t*)s, &str); s += 4; } else if (type == 'i') { kputsn("i:", 2, &str); kputw(*(int32_t*)s, &str); s += 4; } else if (type == 'f') { ksprintf(&str, "f:%g", *(float*)s); s += 4; } else if (type == 'd') { ksprintf(&str, "d:%lg", *(double*)s); s += 8; } else if (type == 'Z' || type == 'H') { kputc(type, &str); kputc(':', &str); while (*s) kputc(*s++, &str); ++s; } else if (type == 'B') { uint8_t sub_type = *(s++); int32_t n; memcpy(&n, s, 4); s += 4; // no point to the start of the array kputc(type, &str); kputc(':', &str); kputc(sub_type, &str); // write the typing for (i = 0; i < n; ++i) { kputc(',', &str); if ('c' == sub_type || 'c' == sub_type) { kputw(*(int8_t*)s, &str); ++s; } else if ('C' == sub_type) { kputw(*(uint8_t*)s, &str); ++s; } else if ('s' == sub_type) { kputw(*(int16_t*)s, &str); s += 2; } else if ('S' == sub_type) { kputw(*(uint16_t*)s, &str); s += 2; } else if ('i' == sub_type) { kputw(*(int32_t*)s, &str); s += 4; } else if ('I' == sub_type) { kputuw(*(uint32_t*)s, &str); s += 4; } else if ('f' == sub_type) { ksprintf(&str, "%g", *(float*)s); s += 4; } } } } return str.s; } char *bam_format1(const bam_header_t *header, const bam1_t *b) { return bam_format1_core(header, b, BAM_OFDEC); } void bam_view1(const bam_header_t *header, const bam1_t *b) { char *s = bam_format1(header, b); puts(s); free(s); } int bam_validate1(const bam_header_t *header, const bam1_t *b) { char *s; if (b->core.tid < -1 || b->core.mtid < -1) return 0; if (header && (b->core.tid >= header->n_targets || b->core.mtid >= header->n_targets)) return 0; if (b->data_len < b->core.l_qname) return 0; s = memchr(bam1_qname(b), '\0', b->core.l_qname); if (s != &bam1_qname(b)[b->core.l_qname-1]) return 0; // FIXME: Other fields could also be checked, especially the auxiliary data return 1; } // FIXME: we should also check the LB tag associated with each alignment const char *bam_get_library(bam_header_t *h, const bam1_t *b) { const uint8_t *rg; if (h->dict == 0) h->dict = sam_header_parse2(h->text); if (h->rg2lib == 0) h->rg2lib = sam_header2tbl(h->dict, "RG", "ID", "LB"); rg = bam_aux_get(b, "RG"); return (rg == 0)? 0 : sam_tbl_get(h->rg2lib, (const char*)(rg + 1)); } /************ * Remove B * ************/ int bam_remove_B(bam1_t *b) { int i, j, end_j, k, l, no_qual; uint32_t *cigar, *new_cigar; uint8_t *seq, *qual, *p; // test if removal is necessary if (b->core.flag & BAM_FUNMAP) return 0; // unmapped; do nothing cigar = bam1_cigar(b); for (k = 0; k < b->core.n_cigar; ++k) if (bam_cigar_op(cigar[k]) == BAM_CBACK) break; if (k == b->core.n_cigar) return 0; // no 'B' if (bam_cigar_op(cigar[0]) == BAM_CBACK) goto rmB_err; // cannot be removed // allocate memory for the new CIGAR if (b->data_len + (b->core.n_cigar + 1) * 4 > b->m_data) { // not enough memory b->m_data = b->data_len + b->core.n_cigar * 4; kroundup32(b->m_data); b->data = (uint8_t*)realloc(b->data, b->m_data); cigar = bam1_cigar(b); // after realloc, cigar may be changed } new_cigar = (uint32_t*)(b->data + (b->m_data - b->core.n_cigar * 4)); // from the end of b->data // the core loop seq = bam1_seq(b); qual = bam1_qual(b); no_qual = (qual[0] == 0xff); // test whether base quality is available i = j = 0; end_j = -1; for (k = l = 0; k < b->core.n_cigar; ++k) { int op = bam_cigar_op(cigar[k]); int len = bam_cigar_oplen(cigar[k]); if (op == BAM_CBACK) { // the backward operation int t, u; if (k == b->core.n_cigar - 1) break; // ignore 'B' at the end of CIGAR if (len > j) goto rmB_err; // an excessively long backward for (t = l - 1, u = 0; t >= 0; --t) { // look back int op1 = bam_cigar_op(new_cigar[t]); int len1 = bam_cigar_oplen(new_cigar[t]); if (bam_cigar_type(op1)&1) { // consume the query if (u + len1 >= len) { // stop new_cigar[t] -= (len - u) << BAM_CIGAR_SHIFT; break; } else u += len1; } } if (bam_cigar_oplen(new_cigar[t]) == 0) --t; // squeeze out the zero-length operation l = t + 1; end_j = j; j -= len; } else { // other CIGAR operations new_cigar[l++] = cigar[k]; if (bam_cigar_type(op)&1) { // consume the query if (i != j) { // no need to copy if i == j int u, c, c0; for (u = 0; u < len; ++u) { // construct the consensus c = bam1_seqi(seq, i+u); if (j + u < end_j) { // in an overlap c0 = bam1_seqi(seq, j+u); if (c != c0) { // a mismatch; choose the better base if (qual[j+u] < qual[i+u]) { // the base in the 2nd segment is better bam1_seq_seti(seq, j+u, c); qual[j+u] = qual[i+u] - qual[j+u]; } else qual[j+u] -= qual[i+u]; // the 1st is better; reduce base quality } else qual[j+u] = qual[j+u] > qual[i+u]? qual[j+u] : qual[i+u]; } else { // not in an overlap; copy over bam1_seq_seti(seq, j+u, c); qual[j+u] = qual[i+u]; } } } i += len, j += len; } } } if (no_qual) qual[0] = 0xff; // in very rare cases, this may be modified // merge adjacent operations if possible for (k = 1; k < l; ++k) if (bam_cigar_op(new_cigar[k]) == bam_cigar_op(new_cigar[k-1])) new_cigar[k] += new_cigar[k-1] >> BAM_CIGAR_SHIFT << BAM_CIGAR_SHIFT, new_cigar[k-1] &= 0xf; // kill zero length operations for (k = i = 0; k < l; ++k) if (new_cigar[k] >> BAM_CIGAR_SHIFT) new_cigar[i++] = new_cigar[k]; l = i; // update b memcpy(cigar, new_cigar, l * 4); // set CIGAR p = b->data + b->core.l_qname + l * 4; memmove(p, seq, (j+1)>>1); p += (j+1)>>1; // set SEQ memmove(p, qual, j); p += j; // set QUAL memmove(p, bam1_aux(b), b->l_aux); p += b->l_aux; // set optional fields b->core.n_cigar = l, b->core.l_qseq = j; // update CIGAR length and query length b->data_len = p - b->data; // update record length return 0; rmB_err: b->core.flag |= BAM_FUNMAP; return -1; }