#include "sam_header.h"
#include "sam.h"
#include "bam.h"
+#include "faidx.h"
+
+bam_header_t *bam_header_dup(const bam_header_t *h0); /*in sam.c*/
static void replace_cigar(bam1_t *b, int n, uint32_t *cigar)
{
assert(length == s->l);
}
-int bam_pad2unpad(samfile_t *in, samfile_t *out)
+int load_unpadded_ref(faidx_t *fai, char *ref_name, int ref_len, kstring_t *seq)
+{
+ char base;
+ char *fai_ref = 0;
+ int fai_ref_len = 0, k;
+
+ fai_ref = fai_fetch(fai, ref_name, &fai_ref_len);
+ if (fai_ref_len != ref_len) {
+ fprintf(stderr, "[depad] ERROR: FASTA sequence %s length %i, expected %i\n", ref_name, fai_ref_len, ref_len);
+ free(fai_ref);
+ return -1;
+ }
+ ks_resize(seq, ref_len);
+ seq->l = 0;
+ for (k = 0; k < ref_len; ++k) {
+ base = fai_ref[k];
+ if (base == '-' || base == '*') {
+ // Map gaps to null to match unpad_seq function
+ seq->s[seq->l++] = 0;
+ } else {
+ int i = bam_nt16_table[(int)base];
+ if (i == 0 || i==16) { // Equals maps to 0, anything unexpected to 16
+ fprintf(stderr, "[depad] ERROR: Invalid character %c (ASCII %i) in FASTA sequence %s\n", base, (int)base, ref_name);
+ free(fai_ref);
+ return -1;
+ }
+ seq->s[seq->l++] = i;
+ }
+ }
+ assert(ref_len == seq->l);
+ free(fai_ref);
+ return 0;
+}
+
+int get_unpadded_len(faidx_t *fai, char *ref_name, int padded_len)
+{
+ char base;
+ char *fai_ref = 0;
+ int fai_ref_len = 0, k;
+ int bases=0, gaps=0;
+
+ fai_ref = fai_fetch(fai, ref_name, &fai_ref_len);
+ if (fai_ref_len != padded_len) {
+ fprintf(stderr, "[depad] ERROR: FASTA sequence '%s' length %i, expected %i\n", ref_name, fai_ref_len, padded_len);
+ free(fai_ref);
+ return -1;
+ }
+ for (k = 0; k < padded_len; ++k) {
+ //fprintf(stderr, "[depad] checking base %i of %i or %i\n", k+1, ref_len, strlen(fai_ref));
+ base = fai_ref[k];
+ if (base == '-' || base == '*') {
+ gaps += 1;
+ } else {
+ int i = bam_nt16_table[(int)base];
+ if (i == 0 || i==16) { // Equals maps to 0, anything unexpected to 16
+ fprintf(stderr, "[depad] ERROR: Invalid character %c (ASCII %i) in FASTA sequence '%s'\n", base, (int)base, ref_name);
+ free(fai_ref);
+ return -1;
+ }
+ bases += 1;
+ }
+ }
+ free(fai_ref);
+ assert (padded_len == bases + gaps);
+ return bases;
+}
+
+inline int * update_posmap(int *posmap, kstring_t ref)
+{
+ int i, k;
+ posmap = realloc(posmap, ref.m * sizeof(int));
+ for (i = k = 0; i < ref.l; ++i) {
+ posmap[i] = k;
+ if (ref.s[i]) ++k;
+ }
+ return posmap;
+}
+
+int bam_pad2unpad(samfile_t *in, samfile_t *out, faidx_t *fai)
{
- bam_header_t *h;
- bam1_t *b;
+ bam_header_t *h = 0;
+ bam1_t *b = 0;
kstring_t r, q;
int r_tid = -1;
uint32_t *cigar2 = 0;
uint32_t *cigar = bam1_cigar(b);
n2 = 0;
if (b->core.pos == 0 && b->core.tid >= 0 && strcmp(bam1_qname(b), h->target_name[b->core.tid]) == 0) {
- int i, k;
- /*
- fprintf(stderr, "[depad] Found embedded reference %s\n", bam1_qname(b));
- */
+ // fprintf(stderr, "[depad] Found embedded reference '%s'\n", bam1_qname(b));
r_tid = b->core.tid;
unpad_seq(b, &r);
if (h->target_len[r_tid] != r.l) {
- fprintf(stderr, "[depad] ERROR: (Padded) length of %s is %i in BAM header, but %ld in embedded reference\n", bam1_qname(b), h->target_len[r_tid], r.l);
+ fprintf(stderr, "[depad] ERROR: (Padded) length of '%s' is %d in BAM header, but %ld in embedded reference\n", bam1_qname(b), h->target_len[r_tid], r.l);
return -1;
}
+ if (fai) {
+ // Check the embedded reference matches the FASTA file
+ if (load_unpadded_ref(fai, h->target_name[b->core.tid], h->target_len[b->core.tid], &q)) {
+ fprintf(stderr, "[depad] ERROR: Failed to load embedded reference '%s' from FASTA\n", h->target_name[b->core.tid]);
+ return -1;
+ }
+ assert(r.l == q.l);
+ int i;
+ for (i = 0; i < r.l; ++i) {
+ if (r.s[i] != q.s[i]) {
+ // Show gaps as ASCII 45
+ fprintf(stderr, "[depad] ERROR: Embedded sequence and reference FASTA don't match for %s base %i, '%c' vs '%c'\n",
+ h->target_name[b->core.tid], i+1,
+ r.s[i] ? bam_nt16_rev_table[(int)r.s[i]] : 45,
+ q.s[i] ? bam_nt16_rev_table[(int)q.s[i]] : 45);
+ return -1;
+ }
+ }
+ }
write_cigar(cigar2, n2, m2, bam_cigar_gen(b->core.l_qseq, BAM_CMATCH));
replace_cigar(b, n2, cigar2);
- posmap = realloc(posmap, r.m * sizeof(int));
- for (i = k = 0; i < r.l; ++i) {
- posmap[i] = k;
- if (r.s[i]) ++k;
- }
+ posmap = update_posmap(posmap, r);
} else if (b->core.n_cigar > 0) {
int i, k, op;
if (b->core.tid < 0) {
- fprintf(stderr, "[depad] ERROR: Read %s has CIGAR but no RNAME\n", bam1_qname(b));
+ fprintf(stderr, "[depad] ERROR: Read '%s' has CIGAR but no RNAME\n", bam1_qname(b));
return -1;
- } else if (b->core.tid != r_tid) {
- fprintf(stderr, "[depad] ERROR: Missing %s embedded reference sequence\n", h->target_name[b->core.tid]);
+ } else if (b->core.tid == r_tid) {
+ ; // good case, reference available
+ //fprintf(stderr, "[depad] Have ref '%s' for read '%s'\n", h->target_name[b->core.tid], bam1_qname(b));
+ } else if (fai) {
+ if (load_unpadded_ref(fai, h->target_name[b->core.tid], h->target_len[b->core.tid], &r)) {
+ fprintf(stderr, "[depad] ERROR: Failed to load '%s' from reference FASTA\n", h->target_name[b->core.tid]);
+ return -1;
+ }
+ posmap = update_posmap(posmap, r);
+ r_tid = b->core.tid;
+ // fprintf(stderr, "[depad] Loaded %s from FASTA file\n", h->target_name[b->core.tid]);
+ } else {
+ fprintf(stderr, "[depad] ERROR: Missing %s embedded reference sequence (and no FASTA file)\n", h->target_name[b->core.tid]);
return -1;
}
unpad_seq(b, &q);
n2 = k;
replace_cigar(b, n2, cigar2);
b->core.pos = posmap[b->core.pos];
+ if (b->core.mtid < 0 || b->core.mpos < 0) {
+ /* Nice case, no mate to worry about*/
+ // fprintf(stderr, "[depad] Read '%s' mate not mapped\n", bam1_qname(b));
+ /* TODO - Warning if FLAG says mate should be mapped? */
+ /* Clean up funny input where mate position is given but mate reference is missing: */
+ b->core.mtid = -1;
+ b->core.mpos = -1;
+ } else if (b->core.mtid == b->core.tid) {
+ /* Nice case, same reference */
+ // fprintf(stderr, "[depad] Read '%s' mate mapped to same ref\n", bam1_qname(b));
+ b->core.mpos = posmap[b->core.mpos];
+ } else {
+ /* Nasty case, Must load alternative posmap */
+ // fprintf(stderr, "[depad] Loading reference '%s' temporarily\n", h->target_name[b->core.mtid]);
+ if (!fai) {
+ fprintf(stderr, "[depad] ERROR: Needed reference %s sequence for mate (and no FASTA file)\n", h->target_name[b->core.mtid]);
+ return -1;
+ }
+ /* Temporarily load the other reference sequence */
+ if (load_unpadded_ref(fai, h->target_name[b->core.mtid], h->target_len[b->core.mtid], &r)) {
+ fprintf(stderr, "[depad] ERROR: Failed to load '%s' from reference FASTA\n", h->target_name[b->core.mtid]);
+ return -1;
+ }
+ posmap = update_posmap(posmap, r);
+ b->core.mpos = posmap[b->core.mpos];
+ /* Restore the reference and posmap*/
+ if (load_unpadded_ref(fai, h->target_name[b->core.tid], h->target_len[b->core.tid], &r)) {
+ fprintf(stderr, "[depad] ERROR: Failed to load '%s' from reference FASTA\n", h->target_name[b->core.tid]);
+ return -1;
+ }
+ posmap = update_posmap(posmap, r);
+ }
}
samwrite(out, b);
}
return ret;
}
+bam_header_t * fix_header(bam_header_t *old, faidx_t *fai)
+{
+ int i = 0, unpadded_len = 0;
+ bam_header_t *header = 0 ;
+
+ header = bam_header_dup(old);
+ for (i = 0; i < old->n_targets; ++i) {
+ unpadded_len = get_unpadded_len(fai, old->target_name[i], old->target_len[i]);
+ if (unpadded_len < 0) {
+ fprintf(stderr, "[depad] ERROR getting unpadded length of '%s', padded length %i\n", old->target_name[i], old->target_len[i]);
+ } else {
+ header->target_len[i] = unpadded_len;
+ //fprintf(stderr, "[depad] Recalculating '%s' length %i -> %i\n", old->target_name[i], old->target_len[i], header->target_len[i]);
+ }
+ }
+ /* Duplicating the header allocated new buffer for header string */
+ /* After modifying the @SQ lines it will only get smaller, since */
+ /* the LN entries will be the same or shorter, and we'll remove */
+ /* any MD entries (MD5 checksums). */
+ assert(strlen(old->text) == strlen(header->text));
+ assert (0==strcmp(old->text, header->text));
+ const char *text;
+ text = old->text;
+ header->text[0] = '\0'; /* Resuse the allocated buffer */
+ char * newtext = header->text;
+ char * end=NULL;
+ while (text[0]=='@') {
+ end = strchr(text, '\n');
+ assert(end != 0);
+ if (text[1]=='S' && text[2]=='Q' && text[3]=='\t') {
+ /* TODO - edit the @SQ line here to remove MD and fix LN. */
+ /* For now just remove the @SQ line, and samtools will */
+ /* automatically generate a minimal replacement with LN. */
+ /* However, that discards any other tags like AS, SP, UR. */
+ //fprintf(stderr, "[depad] Removing @SQ line\n");
+ } else {
+ /* Copy this line to the new header */
+ strncat(newtext, text, end - text + 1);
+ }
+ text = end + 1;
+ }
+ assert (text[0]=='\0');
+ /* Check we didn't overflow the buffer */
+ assert (strlen(header->text) <= strlen(old->text));
+ if (strlen(header->text) < header->l_text) {
+ //fprintf(stderr, "[depad] Reallocating header buffer\n");
+ assert (newtext == header->text);
+ newtext = malloc(strlen(header->text) + 1);
+ strcpy(newtext, header->text);
+ free(header->text);
+ header->text = newtext;
+ header->l_text = strlen(newtext);
+ }
+ //fprintf(stderr, "[depad] Here is the new header (pending @SQ lines),\n\n%s\n(end)\n", header->text);
+ return header;
+}
+
static int usage(int is_long_help);
int main_pad2unpad(int argc, char *argv[])
{
samfile_t *in = 0, *out = 0;
+ bam_header_t *h = 0;
+ faidx_t *fai = 0;
int c, is_bamin = 1, compress_level = -1, is_bamout = 1, is_long_help = 0;
char in_mode[5], out_mode[5], *fn_out = 0, *fn_list = 0, *fn_ref = 0;
int ret=0;
case 'o': fn_out = strdup(optarg); break;
case 'u': assert(is_bamout == 1); compress_level = 0; break;
case '1': assert(is_bamout == 1); compress_level = 1; break;
- case 'T': fn_ref = strdup(optarg); is_bamin = 0; break;
+ case 'T': fn_ref = strdup(optarg); break;
case '?': is_long_help = 1; break;
default: return usage(is_long_help);
}
}
// Load FASTA reference (also needed for SAM -> BAM if missing header)
- if (fn_ref) fn_list = samfaipath(fn_ref);
-
+ if (fn_ref) {
+ fn_list = samfaipath(fn_ref);
+ fai = fai_load(fn_ref);
+ }
// open file handlers
if ((in = samopen(argv[optind], in_mode, fn_list)) == 0) {
- fprintf(stderr, "[depad] fail to open \"%s\" for reading.\n", argv[optind]);
+ fprintf(stderr, "[depad] failed to open \"%s\" for reading.\n", argv[optind]);
ret = 1;
goto depad_end;
}
if (in->header == 0) {
- fprintf(stderr, "[depad] fail to read the header from \"%s\".\n", argv[optind]);
+ fprintf(stderr, "[depad] failed to read the header from \"%s\".\n", argv[optind]);
ret = 1;
goto depad_end;
}
- /* TODO - The reference sequence lengths in the BAM + SAM headers should be updated */
- if ((out = samopen(fn_out? fn_out : "-", out_mode, in->header)) == 0) {
- fprintf(stderr, "[depad] fail to open \"%s\" for writing.\n", fn_out? fn_out : "standard output");
+ if (in->header->text == 0 || in->header->l_text == 0) {
+ fprintf(stderr, "[depad] Warning - failed to read any header text from \"%s\".\n", argv[optind]);
+ assert (0 == in->header->l_text);
+ assert (0 == in->header->text);
+ }
+ if (fn_ref) {
+ h = fix_header(in->header, fai);
+ } else {
+ fprintf(stderr, "[depad] Warning - reference lengths will not be corrected without FASTA reference\n");
+ h = in->header;
+ }
+ if ((out = samopen(fn_out? fn_out : "-", out_mode, h)) == 0) {
+ fprintf(stderr, "[depad] failed to open \"%s\" for writing.\n", fn_out? fn_out : "standard output");
ret = 1;
goto depad_end;
}
// Do the depad
- ret = bam_pad2unpad(in, out);
+ ret = bam_pad2unpad(in, out, fai);
depad_end:
// close files, free and return
- free(fn_list); free(fn_out);
+ if (fai) fai_destroy(fai);
+ if (h != in->header) bam_header_destroy(h);
samclose(in);
samclose(out);
+ free(fn_list); free(fn_out);
return ret;
}
projects / samtools.git / blobdiff