int bam_flagstat(int argc, char *argv[])
{
bamFile fp;
bam_header_t *header;
bam_flagstat_t *s;
if (argc == optind) {
fprintf(pysamerr, "Usage: samtools flagstat <in.bam>\n");
return 1;
}
fp = strcmp(argv[optind], "-")? bam_open(argv[optind], "r") : bam_dopen(fileno(stdin), "r");
assert(fp);
header = bam_header_read(fp);
s = bam_flagstat_core(fp);
printf("%lld + %lld in total (QC-passed reads + QC-failed reads)\n", s->n_reads[0], s->n_reads[1]);
printf("%lld + %lld duplicates\n", s->n_dup[0], s->n_dup[1]);
printf("%lld + %lld mapped (%.2f%%:%.2f%%)\n", s->n_mapped[0], s->n_mapped[1], (float)s->n_mapped[0] / s->n_reads[0] * 100.0, (float)s->n_mapped[1] / s->n_reads[1] * 100.0);
printf("%lld + %lld paired in sequencing\n", s->n_pair_all[0], s->n_pair_all[1]);
printf("%lld + %lld read1\n", s->n_read1[0], s->n_read1[1]);
printf("%lld + %lld read2\n", s->n_read2[0], s->n_read2[1]);
printf("%lld + %lld properly paired (%.2f%%:%.2f%%)\n", s->n_pair_good[0], s->n_pair_good[1], (float)s->n_pair_good[0] / s->n_pair_all[0] * 100.0, (float)s->n_pair_good[1] / s->n_pair_all[1] * 100.0);
printf("%lld + %lld with itself and mate mapped\n", s->n_pair_map[0], s->n_pair_map[1]);
printf("%lld + %lld singletons (%.2f%%:%.2f%%)\n", s->n_sgltn[0], s->n_sgltn[1], (float)s->n_sgltn[0] / s->n_pair_all[0] * 100.0, (float)s->n_sgltn[1] / s->n_pair_all[1] * 100.0);
printf("%lld + %lld with mate mapped to a different chr\n", s->n_diffchr[0], s->n_diffchr[1]);
printf("%lld + %lld with mate mapped to a different chr (mapQ>=5)\n", s->n_diffhigh[0], s->n_diffhigh[1]);
free(s);
bam_header_destroy(header);
bam_close(fp);
return 0;
}
samfile_t* samopen_in(quip_reader_t reader, void* reader_data, bool binary, void* aux)
{
samfile_t *fp;
fp = (samfile_t*)calloc(1, sizeof(samfile_t));
fp->type |= TYPE_READ;
if (binary) { // binary
fp->type |= TYPE_BAM;
fp->x.bam = bam_open_in(reader, reader_data);
if (fp->x.bam == 0) goto open_err_ret;
fp->header = bam_header_read(fp->x.bam);
} else { // text
fp->x.tamr = sam_open_in(reader, reader_data);
if (fp->x.tamr == 0) goto open_err_ret;
fp->header = sam_header_read(fp->x.tamr);
if (fp->header->n_targets == 0) { // no @SQ fields
if (aux) { // check if aux is present
bam_header_t *textheader = fp->header;
fp->header = sam_header_read2((const char*)aux);
if (fp->header == 0) goto open_err_ret;
append_header_text(fp->header, textheader->text, textheader->l_text);
bam_header_destroy(textheader);
}
if (fp->header->n_targets == 0 && bam_verbose >= 1)
fprintf(stderr, "[samopen] no @SQ lines in the header.\n");
} else if (bam_verbose >= 2) fprintf(stderr, "[samopen] SAM header is present: %d sequences.\n", fp->header->n_targets);
}
return fp;
open_err_ret:
free(fp);
return 0;
}
// currently, this function ONLY works if each read has one hit
void bam_mating_core(bamFile in, bamFile out)
{
bam_header_t *header;
bam1_t *b[2];
int curr, has_prev, pre_end = 0, cur_end;
kstring_t str;
str.l = str.m = 0; str.s = 0;
header = bam_header_read(in);
bam_header_write(out, header);
b[0] = bam_init1();
b[1] = bam_init1();
curr = 0; has_prev = 0;
while (bam_read1(in, b[curr]) >= 0) {
bam1_t *cur = b[curr], *pre = b[1-curr];
if (cur->core.tid < 0) continue;
cur_end = bam_calend(&cur->core, bam1_cigar(cur));
if (cur_end > (int)header->target_len[cur->core.tid]) cur->core.flag |= BAM_FUNMAP;
if (cur->core.flag & BAM_FSECONDARY) continue; // skip secondary alignments
if (has_prev) {
if (strcmp(bam1_qname(cur), bam1_qname(pre)) == 0) { // identical pair name
cur->core.mtid = pre->core.tid; cur->core.mpos = pre->core.pos;
pre->core.mtid = cur->core.tid; pre->core.mpos = cur->core.pos;
if (pre->core.tid == cur->core.tid && !(cur->core.flag&(BAM_FUNMAP|BAM_FMUNMAP))
&& !(pre->core.flag&(BAM_FUNMAP|BAM_FMUNMAP))) // set TLEN/ISIZE
{
uint32_t cur5, pre5;
cur5 = (cur->core.flag&BAM_FREVERSE)? cur_end : cur->core.pos;
pre5 = (pre->core.flag&BAM_FREVERSE)? pre_end : pre->core.pos;
cur->core.isize = pre5 - cur5; pre->core.isize = cur5 - pre5;
} else cur->core.isize = pre->core.isize = 0;
if (pre->core.flag&BAM_FREVERSE) cur->core.flag |= BAM_FMREVERSE;
else cur->core.flag &= ~BAM_FMREVERSE;
if (cur->core.flag&BAM_FREVERSE) pre->core.flag |= BAM_FMREVERSE;
else pre->core.flag &= ~BAM_FMREVERSE;
if (cur->core.flag & BAM_FUNMAP) { pre->core.flag |= BAM_FMUNMAP; pre->core.flag &= ~BAM_FPROPER_PAIR; }
if (pre->core.flag & BAM_FUNMAP) { cur->core.flag |= BAM_FMUNMAP; cur->core.flag &= ~BAM_FPROPER_PAIR; }
bam_template_cigar(pre, cur, &str);
bam_write1(out, pre);
bam_write1(out, cur);
has_prev = 0;
} else { // unpaired or singleton
pre->core.mtid = -1; pre->core.mpos = -1; pre->core.isize = 0;
if (pre->core.flag & BAM_FPAIRED) {
pre->core.flag |= BAM_FMUNMAP;
pre->core.flag &= ~BAM_FMREVERSE & ~BAM_FPROPER_PAIR;
}
bam_write1(out, pre);
}
} else has_prev = 1;
curr = 1 - curr;
pre_end = cur_end;
}
if (has_prev) bam_write1(out, b[1-curr]);
bam_header_destroy(header);
bam_destroy1(b[0]);
bam_destroy1(b[1]);
free(str.s);
}
void samclose(samfile_t *fp)
{
if (fp == 0) return;
if (fp->header) bam_header_destroy(fp->header);
if (fp->type & 1) bam_close(fp->x.bam);
else if (fp->type == 2) sam_close(fp->x.tamr);
free(fp);
}
samfile_t* samopen_out(quip_writer_t writer, void* writer_data, bool binary, void* aux)
{
samfile_t *fp;
fp = (samfile_t*)calloc(1, sizeof(samfile_t));
fp->header = bam_header_dup((const bam_header_t*)aux);
if (binary) { // binary
fp->type |= TYPE_BAM;
fp->x.bam = bam_open_out(writer, writer_data);
if (fp->x.bam == 0) goto open_err_ret;
bam_header_write(fp->x.bam, fp->header);
} else { // text
// open file
fp->x.tamw.writer = writer;
fp->x.tamw.writer_data = writer_data;
fp->type |= BAM_OFDEC<<2;
// write header
if (aux) {
int i;
bam_header_t *alt;
// parse the header text
alt = bam_header_init();
alt->l_text = fp->header->l_text;
alt->text = fp->header->text;
sam_header_parse(alt);
alt->l_text = 0;
alt->text = 0;
// check if there are @SQ lines in the header
writer(writer_data, (uint8_t*) fp->header->text, fp->header->l_text);
if (alt->n_targets) { // then write the header text without dumping ->target_{name,len}
if (alt->n_targets != fp->header->n_targets && bam_verbose >= 1)
fprintf(stderr, "[samopen] inconsistent number of target sequences. Output the text header.\n");
} else { // then dump ->target_{name,len}
char target_len_str[11];
for (i = 0; i < fp->header->n_targets; ++i) {
writer(writer_data, (uint8_t*) "@SQ\tSN:", 7);
writer(writer_data, (uint8_t*) fp->header->target_name[i], strlen(fp->header->target_name[i]));
writer(writer_data, (uint8_t*) "\tLN:", 4);
snprintf(target_len_str, sizeof(target_len_str), "%"PRIu32, fp->header->target_len[i]);
writer(writer_data, (uint8_t*) target_len_str, strlen(target_len_str));
writer(writer_data, (uint8_t*) "\n", 1);
}
}
bam_header_destroy(alt);
}
}
return fp;
open_err_ret:
free(fp);
return 0;
}
void samclose(samfile_t *fp)
{
if (fp == 0) return;
if (fp->header) bam_header_destroy(fp->header);
if (fp->type & TYPE_BAM) bam_close(fp->x.bam);
else if (fp->type & TYPE_READ) sam_close(fp->x.tamr);
else fclose(fp->x.tamw);
free(fp);
}
/* check match between reference and bam files. prints an error
* message and return non-zero on mismatch
*/
int checkref(char *fasta_file, char *bam_file)
{
int i = -1;
bam_header_t *header;
faidx_t *fai;
char *ref;
int ref_len = -1;
bamFile bam_fp;
if (! file_exists(fasta_file)) {
LOG_FATAL("Fsata file %s does not exist. Exiting...\n", fasta_file);
return 1;
}
if (0 != strcmp(bam_file, "-") && ! file_exists(bam_file)) {
LOG_FATAL("BAM file %s does not exist. Exiting...\n", bam_file);
return 1;
}
bam_fp = strcmp(bam_file, "-") == 0 ? bam_dopen(fileno(stdin), "r") : bam_open(bam_file, "r");
header = bam_header_read(bam_fp);
if (!header) {
LOG_FATAL("Failed to read BAM header from %s\n", bam_file);
return 1;
}
fai = fai_load(fasta_file);
if (!fai) {
LOG_FATAL("Failed to fasta index for %s\n", fasta_file);
return 1;
}
for (i=0; i < header->n_targets; i++) {
LOG_DEBUG("BAM header target %d of %d: name=%s len=%d\n",
i+1, header->n_targets, header->target_name[i], header->target_len[i]);
ref = faidx_fetch_seq(fai, header->target_name[i],
0, 0x7fffffff, &ref_len);
if (NULL == ref) {
LOG_FATAL("Failed to fetch sequence %s from fasta file\n", header->target_name[i]);
return -1;
}
if (header->target_len[i] != ref_len) {
LOG_FATAL("Sequence length mismatch for sequence %s (%dbp in fasta; %dbp in bam)\n",
header->target_name[i], header->target_len[i], ref_len);
return -1;
}
free(ref);
}
fai_destroy(fai);
bam_header_destroy(header);
bam_close(bam_fp);
return 0;
}
void TGM_BamHeaderFree(TGM_BamHeader* pBamHeader)
{
if (pBamHeader != NULL)
{
free(pBamHeader->pMD5s);
bam_header_destroy(pBamHeader->pOrigHeader);
free(pBamHeader);
}
}
int bam_pad2unpad(bamFile in, bamFile out)
{
bam_header_t *h;
bam1_t *b;
kstring_t r, q;
uint32_t *cigar2 = 0;
int n2 = 0, m2 = 0, *posmap = 0;
h = bam_header_read(in);
bam_header_write(out, h);
b = bam_init1();
r.l = r.m = q.l = q.m = 0; r.s = q.s = 0;
while (bam_read1(in, b) >= 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;
unpad_seq(b, &r);
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; // note that a read should NOT start at a padding
if (r.s[i]) ++k;
}
} else {
int i, k, op;
unpad_seq(b, &q);
if (bam_cigar_op(cigar[0]) == BAM_CSOFT_CLIP) write_cigar(cigar2, n2, m2, cigar[0]);
for (i = 0, k = b->core.pos; i < q.l; ++i, ++k)
q.s[i] = q.s[i]? (r.s[k]? BAM_CMATCH : BAM_CINS) : (r.s[k]? BAM_CDEL : BAM_CPAD);
for (i = k = 1, op = q.s[0]; i < q.l; ++i) {
if (op != q.s[i]) {
write_cigar(cigar2, n2, m2, bam_cigar_gen(k, op));
op = q.s[i]; k = 1;
} else ++k;
}
write_cigar(cigar2, n2, m2, bam_cigar_gen(k, op));
if (bam_cigar_op(cigar[b->core.n_cigar-1]) == BAM_CSOFT_CLIP) write_cigar(cigar2, n2, m2, cigar[b->core.n_cigar-1]);
for (i = 2; i < n2; ++i)
if (bam_cigar_op(cigar2[i]) == BAM_CMATCH && bam_cigar_op(cigar2[i-1]) == BAM_CPAD && bam_cigar_op(cigar2[i-2]) == BAM_CMATCH)
cigar2[i] += cigar2[i-2], cigar2[i-2] = cigar2[i-1] = 0;
for (i = k = 0; i < n2; ++i)
if (cigar2[i]) cigar2[k++] = cigar2[i];
n2 = k;
replace_cigar(b, n2, cigar2);
b->core.pos = posmap[b->core.pos];
}
bam_write1(out, b);
}
free(r.s); free(q.s); free(posmap);
bam_destroy1(b);
bam_header_destroy(h);
return 0;
}
bwa_seqio_t *bwa_bam_open(const char *fn, int which) {
bwa_seqio_t *bs;
bam_header_t *h;
bs = (bwa_seqio_t*) calloc(1, sizeof(bwa_seqio_t));
bs->is_bam = 1;
bs->which = which;
bs->fp = bam_open(fn, "r");
h = bam_header_read(bs->fp);
bam_header_destroy(h);
return bs;
}
int main(int argc, char** argv)
{
if(argc < 3) {
printf("No input nor output files provided");
return -1;
}
bamFile in = bam_open(argv[1], "r");
bam_header_t* header;
if (in == NULL) {
printf("opening input file failed");
return -1;
}
bam1_t* b = bam_init1();
bamFile out = bam_open(argv[2], "w");
if (out == NULL) {
printf("opening input file failed");
return -1;
}
header = bam_header_read(in);
if(bam_header_write(out, header) < 0) {
printf("writing header failed");
}
long nextPrunedId;
if(!scanf ("%lu", &nextPrunedId)) {
printf("warning: no ids provided");
return -1;
}
long id = 0;
while (bam_read1(in, b) >= 0) {
// write BAM back
if (nextPrunedId != id++) {
bam_write1(out, b);
} else {
// fprintf(stderr, "pruning: id: %lu, pos: %d, length: %d\n", nextPrunedId, b->core.pos, b->core.l_qseq);
if(!scanf ("%lu", &nextPrunedId)) {
break;
}
}
}
// closing all resources
bam_header_destroy(header);
bam_close(in);
bam_close(out);
bam_destroy1(b);
return 0;
}
bwa_seqio_t *bwa_bam_open(const char *fn, int which)
{
bwa_seqio_t *bs;
bam_header_t *h;
bs = (bwa_seqio_t*)calloc(1, sizeof(bwa_seqio_t));
bs->is_bam = 1;
bs->which = which;
bs->fp = bam_open(fn, "r");
if (0 == bs->fp) err_fatal_simple("Couldn't open bam file");
h = bam_header_read(bs->fp);
bam_header_destroy(h);
return bs;
}
bwa_seqio_t *bwa_bam_open(const char *fn, int which, char **saif,
gap_opt_t *o0, bam_header_t **hh)
{
int c, b=0;
bwa_seqio_t *bs;
bam_header_t *h;
bs = (bwa_seqio_t*)calloc(1, sizeof(bwa_seqio_t));
bs->is_bam = 1;
bs->which = which;
bs->fp = (fn[0]!='-' || fn[1]) ? bam_open(fn, "r") : bam_dopen(0, "r") ;
h = bam_header_read(bs->fp);
if(hh) *hh=h;
else bam_header_destroy(h);
if( saif ) for(c=0; c!=3; ++c)
{
gap_opt_t opt;
if( saif[c] ) {
bs->sai[c] = xopen(saif[c], "r");
if( 1 > fread(&opt, sizeof(gap_opt_t), 1, bs->sai[c]) )
{
fclose(bs->sai[c]);
bs->sai[c] = 0;
}
opt.n_threads=o0->n_threads;
if(o0) {
if(b) {
opt.mode=o0->mode;
if( memcmp(o0, &opt, sizeof(gap_opt_t)) ) {
fprintf( stderr, "[bwa_bam_open] options from sai file \"%s\" conflict with others.\n", saif[c] ) ;
exit(1);
}
fprintf( stderr, "[bwa_bam_open] options from sai file \"%s\" match.\n", saif[c] ) ;
}
else {
fprintf( stderr, "[bwa_bam_open] recovered options from sai file \"%s\".\n", saif[c] ) ;
memcpy(o0, &opt, sizeof(gap_opt_t));
b=1;
}
}
}
}
return bs;
}
void convert_sam_to_bam(char* sam_input, char* bam_input) {
bam1_t* bam_p = bam_init1();
LOG_DEBUG("CONVERT-START: sam to bam\n");
//open SAM file for read
if (time_flag) {
start_timer(t1_convert);
}
tamFile sam_fd = sam_open(sam_input);
//open BAM file for write
bam_file_t* bam_file_p = bam_fopen_mode(bam_input, NULL, "w");
//read header from SAM file
bam_header_t* bam_header_p = sam_header_read(sam_fd);
//write header to BAM file
bam_header_write(bam_file_p->bam_fd, bam_header_p);
//write alignments to BAM file
while (sam_read1(sam_fd, bam_header_p, bam_p) > 0) {
bam_write1(bam_file_p->bam_fd, bam_p);
num_alignments++;
}
//close BAM and SAM files, free bam alignment and bam file object
bam_fclose(bam_file_p);
sam_close(sam_fd);
bam_header_destroy(bam_header_p);
bam_destroy1(bam_p);
if (time_flag) {
stop_timer(t1_convert, t2_convert, convert_time);
}
//number_of_batchs = 1, convention value for statistics (not real batch)
number_of_batchs = 1;
}
bwa_seqio_t *bwa_bam_open(const char *fn, int which)
{
bwa_seqio_t *bs;
#ifndef USE_HTSLIB
bam_header_t *h;
#endif
bs = (bwa_seqio_t*)calloc(1, sizeof(bwa_seqio_t));
bs->is_bam = 1;
bs->which = which;
#ifdef USE_HTSLIB
bs->fp = sam_open(fn, "rb");
#else
bs->fp = bam_open(fn, "r");
#endif
if (0 == bs->fp) err_fatal_simple("Couldn't open bam file");
#ifdef USE_HTSLIB
bs->h = sam_hdr_read(bs->fp);
#else
h = bam_header_read(bs->fp);
bam_header_destroy(h);
#endif
return bs;
}
int bam_cat(int nfn, char * const *fn, const bam_header_t *h, const char* outbam)
{
BGZF *fp;
uint8_t *buf;
uint8_t ebuf[BGZF_EMPTY_BLOCK_SIZE];
const int es=BGZF_EMPTY_BLOCK_SIZE;
int i;
fp = strcmp(outbam, "-")? bgzf_open(outbam, "w") : bgzf_fdopen(fileno(stdout), "w");
if (fp == 0) {
fprintf(stderr, "[%s] ERROR: fail to open output file '%s'.\n", __func__, outbam);
return 1;
}
if (h) bam_header_write(fp, h);
buf = (uint8_t*) malloc(BUF_SIZE);
for(i = 0; i < nfn; ++i){
BGZF *in;
bam_header_t *old;
int len,j;
in = strcmp(fn[i], "-")? bgzf_open(fn[i], "r") : bgzf_fdopen(fileno(stdin), "r");
if (in == 0) {
fprintf(stderr, "[%s] ERROR: fail to open file '%s'.\n", __func__, fn[i]);
return -1;
}
if (in->is_write) return -1;
old = bam_header_read(in);
if (h == 0 && i == 0) bam_header_write(fp, old);
if (in->block_offset < in->block_length) {
bgzf_write(fp, in->uncompressed_block + in->block_offset, in->block_length - in->block_offset);
bgzf_flush(fp);
}
j=0;
while ((len = bgzf_raw_read(in, buf, BUF_SIZE)) > 0) {
if(len<es){
int diff=es-len;
if(j==0) {
fprintf(stderr, "[%s] ERROR: truncated file?: '%s'.\n", __func__, fn[i]);
return -1;
}
bgzf_raw_write(fp, ebuf, len);
memcpy(ebuf,ebuf+len,diff);
memcpy(ebuf+diff,buf,len);
} else {
if(j!=0) bgzf_raw_write(fp, ebuf, es);
len-= es;
memcpy(ebuf,buf+len,es);
bgzf_raw_write(fp, buf, len);
}
j=1;
}
/* check final gzip block */
{
const uint8_t gzip1=ebuf[0];
const uint8_t gzip2=ebuf[1];
const uint32_t isize=*((uint32_t*)(ebuf+es-4));
if(((gzip1!=GZIPID1) || (gzip2!=GZIPID2)) || (isize!=0)) {
fprintf(stderr, "[%s] WARNING: Unexpected block structure in file '%s'.", __func__, fn[i]);
fprintf(stderr, " Possible output corruption.\n");
bgzf_raw_write(fp, ebuf, es);
}
}
bam_header_destroy(old);
bgzf_close(in);
}
free(buf);
bgzf_close(fp);
return 0;
}
int main_depth(int argc, char *argv[])
#endif
{
int i, n, tid, beg, end, pos, *n_plp, baseQ = 0, mapQ = 0;
const bam_pileup1_t **plp;
char *reg = 0; // specified region
void *bed = 0; // BED data structure
bam_header_t *h = 0; // BAM header of the 1st input
aux_t **data;
bam_mplp_t mplp;
// parse the command line
while ((n = getopt(argc, argv, "r:b:q:Q:")) >= 0) {
switch (n) {
case 'r': reg = strdup(optarg); break; // parsing a region requires a BAM header
case 'b': bed = bed_read(optarg); break; // BED or position list file can be parsed now
case 'q': baseQ = atoi(optarg); break; // base quality threshold
case 'Q': mapQ = atoi(optarg); break; // mapping quality threshold
}
}
if (optind == argc) {
fprintf(stderr, "Usage: bam2depth [-r reg] [-q baseQthres] [-Q mapQthres] [-b in.bed] <in1.bam> [...]\n");
return 1;
}
// initialize the auxiliary data structures
n = argc - optind; // the number of BAMs on the command line
data = (aux_t **) calloc(n, sizeof(void*)); // data[i] for the i-th input
beg = 0; end = 1<<30; tid = -1; // set the default region
for (i = 0; i < n; ++i) {
bam_header_t *htmp;
data[i] = (aux_t *) calloc(1, sizeof(aux_t));
data[i]->fp = bam_open(argv[optind+i], "r"); // open BAM
data[i]->min_mapQ = mapQ; // set the mapQ filter
htmp = bam_header_read(data[i]->fp); // read the BAM header
if (i == 0) {
h = htmp; // keep the header of the 1st BAM
if (reg) bam_parse_region(h, reg, &tid, &beg, &end); // also parse the region
} else bam_header_destroy(htmp); // if not the 1st BAM, trash the header
if (tid >= 0) { // if a region is specified and parsed successfully
bam_index_t *idx = bam_index_load(argv[optind+i]); // load the index
data[i]->iter = bam_iter_query(idx, tid, beg, end); // set the iterator
bam_index_destroy(idx); // the index is not needed any more; phase out of the memory
}
}
// the core multi-pileup loop
mplp = bam_mplp_init(n, read_bam, (void**)data); // initialization
n_plp = (int*) calloc(n, sizeof(int)); // n_plp[i] is the number of covering reads from the i-th BAM
plp = (bam_pileup1_t **) calloc(n, sizeof(void*)); // plp[i] points to the array of covering reads (internal in mplp)
while (bam_mplp_auto(mplp, &tid, &pos, n_plp, plp) > 0) { // come to the next covered position
if (pos < beg || pos >= end) continue; // out of range; skip
if (bed && bed_overlap(bed, h->target_name[tid], pos, pos + 1) == 0) continue; // not in BED; skip
fputs(h->target_name[tid], stdout); printf("\t%d", pos+1); // a customized printf() would be faster
for (i = 0; i < n; ++i) { // base level filters have to go here
int j, m = 0;
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j; // DON'T modfity plp[][] unless you really know
if (p->is_del || p->is_refskip) ++m; // having dels or refskips at tid:pos
else if (bam1_qual(p->b)[p->qpos] < baseQ) ++m; // low base quality
}
printf("\t%d", n_plp[i] - m); // this the depth to output
}
putchar('\n');
}
free(n_plp); free(plp);
bam_mplp_destroy(mplp);
bam_header_destroy(h);
for (i = 0; i < n; ++i) {
bam_close(data[i]->fp);
if (data[i]->iter) bam_iter_destroy(data[i]->iter);
free(data[i]);
}
free(data); free(reg);
if (bed) bed_destroy(bed);
return 0;
}
int main_depth(int argc, char *argv[])
#endif
{
int i, n, tid, beg, end, pos, *n_plp, baseQ = 0, mapQ = 0, min_len = 0, nfiles;
const bam_pileup1_t **plp;
char *reg = 0; // specified region
void *bed = 0; // BED data structure
char *file_list = NULL, **fn = NULL;
bam_header_t *h = 0; // BAM header of the 1st input
aux_t **data;
bam_mplp_t mplp;
// parse the command line
while ((n = getopt(argc, argv, "r:b:q:Q:l:f:")) >= 0) {
switch (n) {
case 'l': min_len = atoi(optarg); break; // minimum query length
case 'r': reg = strdup(optarg); break; // parsing a region requires a BAM header
case 'b': bed = bed_read(optarg); break; // BED or position list file can be parsed now
case 'q': baseQ = atoi(optarg); break; // base quality threshold
case 'Q': mapQ = atoi(optarg); break; // mapping quality threshold
case 'f': file_list = optarg; break;
}
}
if (optind == argc && !file_list) {
fprintf(stderr, "\n");
fprintf(stderr, "Usage: samtools depth [options] in1.bam [in2.bam [...]]\n");
fprintf(stderr, "Options:\n");
fprintf(stderr, " -b <bed> list of positions or regions\n");
fprintf(stderr, " -f <list> list of input BAM filenames, one per line [null]\n");
fprintf(stderr, " -l <int> minQLen\n");
fprintf(stderr, " -q <int> base quality threshold\n");
fprintf(stderr, " -Q <int> mapping quality threshold\n");
fprintf(stderr, " -r <chr:from-to> region\n");
fprintf(stderr, "\n");
return 1;
}
// initialize the auxiliary data structures
if (file_list)
{
if ( read_file_list(file_list,&nfiles,&fn) ) return 1;
n = nfiles;
argv = fn;
optind = 0;
}
else
n = argc - optind; // the number of BAMs on the command line
data = calloc(n, sizeof(void*)); // data[i] for the i-th input
beg = 0; end = 1<<30; tid = -1; // set the default region
for (i = 0; i < n; ++i) {
bam_header_t *htmp;
data[i] = calloc(1, sizeof(aux_t));
data[i]->fp = bam_open(argv[optind+i], "r"); // open BAM
data[i]->min_mapQ = mapQ; // set the mapQ filter
data[i]->min_len = min_len; // set the qlen filter
htmp = bam_header_read(data[i]->fp); // read the BAM header
if (i == 0) {
h = htmp; // keep the header of the 1st BAM
if (reg) bam_parse_region(h, reg, &tid, &beg, &end); // also parse the region
} else bam_header_destroy(htmp); // if not the 1st BAM, trash the header
if (tid >= 0) { // if a region is specified and parsed successfully
bam_index_t *idx = bam_index_load(argv[optind+i]); // load the index
data[i]->iter = bam_iter_query(idx, tid, beg, end); // set the iterator
bam_index_destroy(idx); // the index is not needed any more; phase out of the memory
}
}
// the core multi-pileup loop
mplp = bam_mplp_init(n, read_bam, (void**)data); // initialization
bam_mplp_set_maxcnt(mplp,2147483647); // set max_depth to int max
n_plp = calloc(n, sizeof(int)); // n_plp[i] is the number of covering reads from the i-th BAM
plp = calloc(n, sizeof(void*)); // plp[i] points to the array of covering reads (internal in mplp)
while (bam_mplp_auto(mplp, &tid, &pos, n_plp, plp) > 0) { // come to the next covered position
if (pos < beg || pos >= end) continue; // out of range; skip
if (bed && bed_overlap(bed, h->target_name[tid], pos, pos + 1) == 0) continue; // not in BED; skip
fputs(h->target_name[tid], stdout); printf("\t%d", pos+1); // a customized printf() would be faster
for (i = 0; i < n; ++i) { // base level filters have to go here
int j, m = 0;
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j; // DON'T modfity plp[][] unless you really know
if (p->is_del || p->is_refskip) ++m; // having dels or refskips at tid:pos
else if (bam1_qual(p->b)[p->qpos] < baseQ) ++m; // low base quality
}
printf("\t%d", n_plp[i] - m); // this the depth to output
}
putchar('\n');
}
free(n_plp); free(plp);
bam_mplp_destroy(mplp);
bam_header_destroy(h);
for (i = 0; i < n; ++i) {
bam_close(data[i]->fp);
if (data[i]->iter) bam_iter_destroy(data[i]->iter);
free(data[i]);
}
free(data); free(reg);
if (bed) bed_destroy(bed);
if ( file_list )
{
for (i=0; i<n; i++) free(fn[i]);
free(fn);
}
return 0;
}
int bam_cat(int nfn, char * const *fn, const bam_header_t *h, const char* outbam)
{
BGZF *fp;
FILE* fp_file;
uint8_t *buf;
uint8_t ebuf[BGZF_EMPTY_BLOCK_SIZE];
const int es=BGZF_EMPTY_BLOCK_SIZE;
int i;
fp = strcmp(outbam, "-")? bgzf_open(outbam, "w") : bgzf_fdopen(_fileno(stdout), "w");
if (fp == 0) {
fprintf(stderr, "[%s] ERROR: fail to open output file '%s'.\n", __FUNCTION__, outbam);
return 1;
}
if (h) bam_header_write(fp, h);
buf = (uint8_t*) malloc(BUF_SIZE);
for(i = 0; i < nfn; ++i){
BGZF *in;
bam_header_t *old;
int len,j;
in = strcmp(fn[i], "-")? bam_open(fn[i], "r") : bam_dopen(_fileno(stdin), "r");
if (in == 0) {
fprintf(stderr, "[%s] ERROR: fail to open file '%s'.\n", __FUNCTION__, fn[i]);
return -1;
}
if (in->open_mode != 'r') return -1;
old = bam_header_read(in);
if (h == 0 && i == 0) bam_header_write(fp, old);
if (in->block_offset < in->block_length) {
bgzf_write(fp, (uint8_t*)in->uncompressed_block + in->block_offset, in->block_length - in->block_offset);
bgzf_flush(fp);
}
j=0;
#ifdef _USE_KNETFILE
fp_file=fp->x.fpw;
while ((len = knet_read(in->x.fpr, buf, BUF_SIZE)) > 0) {
#else
fp_file=fp->file;
while (!feof(in->file) && (len = fread(buf, 1, BUF_SIZE, in->file)) > 0) {
#endif
if(len<es){
int diff=es-len;
if(j==0) {
fprintf(stderr, "[%s] ERROR: truncated file?: '%s'.\n", __FUNCTION__, fn[i]);
return -1;
}
fwrite(ebuf, 1, len, fp_file);
memcpy(ebuf,ebuf+len,diff);
memcpy(ebuf+diff,buf,len);
} else {
if(j!=0) fwrite(ebuf, 1, es, fp_file);
len-= es;
memcpy(ebuf,buf+len,es);
fwrite(buf, 1, len, fp_file);
}
j=1;
}
/* check final gzip block */
{
const uint8_t gzip1=ebuf[0];
const uint8_t gzip2=ebuf[1];
const uint32_t isize=*((uint32_t*)(ebuf+es-4));
if(((gzip1!=GZIPID1) || (gzip2!=GZIPID2)) || (isize!=0)) {
fprintf(stderr, "[%s] WARNING: Unexpected block structure in file '%s'.", __FUNCTION__, fn[i]);
fprintf(stderr, " Possible output corruption.\n");
fwrite(ebuf, 1, es, fp_file);
}
}
bam_header_destroy(old);
bgzf_close(in);
}
free(buf);
bgzf_close(fp);
return 0;
}
int main_cat(int argc, char *argv[])
{
bam_header_t *h = 0;
char *outfn = 0;
int c, ret;
while ((c = getopt(argc, argv, "h:o:")) >= 0) {
switch (c) {
case 'h': {
tamFile fph = sam_open(optarg);
if (fph == 0) {
fprintf(stderr, "[%s] ERROR: fail to read the header from '%s'.\n", __FUNCTION__, argv[1]);
return 1;
}
h = sam_header_read(fph);
sam_close(fph);
break;
}
case 'o': outfn = strdup(optarg); break;
}
}
if (argc - optind < 2) {
fprintf(stderr, "Usage: samtools cat [-h header.sam] [-o out.bam] <in1.bam> <in2.bam> [...]\n");
return 1;
}
ret = bam_cat(argc - optind, argv + optind, h, outfn? outfn : "-");
free(outfn);
return ret;
}
int
tmap_seqs_io_sff2sam_main(int argc, char *argv[])
{
int c, help = 0;
tmap_seqs_io_t *io_in = NULL;
tmap_seqs_t *seqs = NULL;
char **sam_rg = NULL;
int32_t sam_rg_num = 0;
int bidirectional = 0, sam_flowspace_tags = 0;
int out_type = 0;
tmap_sam_io_t *io_out = NULL;
bam_header_t *header = NULL; // BAM Header
int32_t i;
/*
uint8_t *key_seq = NULL;
int key_seq_len = 0;
*/
while((c = getopt(argc, argv, "DGR:Yvh")) >= 0) {
switch(c) {
case 'D': bidirectional = 1; break;
case 'G': break;
case 'R':
sam_rg = tmap_realloc(sam_rg, (1+sam_rg_num) * sizeof(char*), "sam_rg");
sam_rg[sam_rg_num] = tmap_strdup(optarg);
sam_rg_num++;
break;
case 'Y': sam_flowspace_tags = 1; break;
case 'v': tmap_progress_set_verbosity(1); break;
case 'h': help = 1; break;
default: return 1;
}
}
if(1 != argc - optind || 1 == help) {
tmap_file_fprintf(tmap_file_stderr, "Usage: %s %s [-R -Y -v -h] <in.sff>\n", PACKAGE, argv[0]);
return 1;
}
// input
io_in = tmap_seqs_io_init(&argv[optind], 1, TMAP_SEQ_TYPE_SFF, TMAP_FILE_NO_COMPRESSION, 0l, 0l);
// BAM Header
header = tmap_seqs_io_to_bam_header(NULL, io_in, sam_rg, sam_rg_num, argc, argv);
// open the output file
switch(out_type) {
case 0: // SAM
io_out = tmap_sam_io_init2("-", "wh", header);
break;
case 1:
io_out = tmap_sam_io_init2("-", "wb", header);
break;
case 2:
io_out = tmap_sam_io_init2("-", "wbu", header);
break;
default:
tmap_bug();
}
// destroy the BAM Header
bam_header_destroy(header);
header = NULL;
seqs = tmap_seqs_init(TMAP_SEQ_TYPE_SFF);
while(0 < tmap_seqs_io_read(io_in, seqs, io_out->fp->header->header)) {
bam1_t *b = NULL;
tmap_seq_t *seq = seqs->seqs[0];
b = tmap_sam_convert_unmapped(seq, sam_flowspace_tags, bidirectional, NULL,
0, 0, 0,
0, 0, 0,
"\tlq:i:%d\trq:i:%d\tla:i:%d\trq:i:%d",
seq->data.sff->rheader->clip_qual_left,
seq->data.sff->rheader->clip_qual_right,
seq->data.sff->rheader->clip_adapter_left,
seq->data.sff->rheader->clip_adapter_right);
if(samwrite(io_out->fp, b) <= 0) {
tmap_error("Error writing the SAM file", Exit, WriteFileError);
}
bam_destroy1(b);
tmap_seqs_destroy(seqs);
seqs = tmap_seqs_init(TMAP_SEQ_TYPE_SFF);
}
tmap_seqs_destroy(seqs);
// free memory
tmap_seqs_io_destroy(io_in);
tmap_sam_io_destroy(io_out);
for(i=0;i<sam_rg_num;i++) {
free(sam_rg[i]);
}
free(sam_rg);
return 0;
}
int main(int argc, char *argv[]) {
int c;
const char* normal_sample_id = _default_normal_sample_id;
const char* tumor_sample_id = _default_tumor_sample_id;
const char *fn_fa = 0;
pu_data2_t *d = (pu_data2_t*)calloc(1, sizeof(pu_data2_t));
d->min_somatic_qual=15;
d->tid = -1; d->mask = BAM_DEF_MASK; d->mapQ = 0;
d->c = sniper_maqcns_init();
int use_priors = 1;
d->include_loh = 1;
d->include_gor = 1;
d->use_joint_priors = 0;
d->somatic_mutation_rate = 0.01;
const char *output_format = "classic";
while ((c = getopt(argc, argv, "n:t:vf:T:N:r:I:q:Q:pLGJs:F:")) >= 0) {
switch (c) {
case 'f': fn_fa = optarg; break;
case 'T': d->c->theta = atof(optarg); break;
case 'N': d->c->n_hap = atoi(optarg); break;
case 'r': d->c->het_rate = atof(optarg); break;
case 'q': d->mapQ = atoi(optarg); break;
case 'Q': d->min_somatic_qual = atoi(optarg); break;
case 'F': output_format = optarg; break;
case 'p': use_priors = 0; break;
case 'J': d->use_joint_priors = 1; break;
case 's': d->somatic_mutation_rate = atof(optarg); d->use_joint_priors = 1; break;
case 'v': version_info(); exit(0); break;
case 't': tumor_sample_id = optarg; break;
case 'n': normal_sample_id = optarg; break;
case 'L': d->include_loh = 0; break;
case 'G': d->include_gor = 0; break;
default: fprintf(stderr, "Unrecognizd option '-%c'.\n", c); return 1;
}
}
if (optind == argc) {
usage(argv[0], d);
sniper_maqcns_destroy(d->c); free(d);
return 1;
}
if (fn_fa) {
d->fai = fai_load(fn_fa);
}
else {
fprintf(stderr, "You MUST specify a reference sequence. It isn't optional.\n");
sniper_maqcns_destroy(d->c);
free(d);
exit(1);
}
if(d->use_joint_priors) {
fprintf(stderr,"Using priors accounting for somatic mutation rate. Prior probability of a somatic mutation is %f\n",d->somatic_mutation_rate);
make_joint_prior(d->somatic_mutation_rate);
}
sniper_maqcns_prepare(d->c);
fprintf(stderr,"Preparing to snipe some somatics\n");
if(use_priors) {
fprintf(stderr,"Using prior probabilities\n");
makeSoloPrior();
}
bamFile fp1, fp2;
qAddTableInit();
fp1 = (strcmp(argv[optind], "-") == 0)? bam_dopen(fileno(stdin), "r") : bam_open(argv[optind], "r");
fprintf(stderr, "Normal bam is %s\n", argv[optind+1]);
fprintf(stderr, "Tumor bam is %s\n", argv[optind]);
d->h1 = bam_header_read(fp1);
sam_header_parse_rg(d->h1);
fp2 = bam_open(argv[optind+1], "r");
d->h2 = bam_header_read(fp2);
sam_header_parse_rg(d->h2);
FILE* snp_fh = fopen(argv[optind+2], "w");
/* this will exit if the format name is invalid */
output_formatter_t fmt = output_formatter_create(output_format, snp_fh);
d->output_formatter = &fmt;
if(snp_fh) {
header_data_t hdr;
hdr.refseq = fn_fa;
hdr.normal_sample_id = normal_sample_id;
hdr.tumor_sample_id = tumor_sample_id;
d->output_formatter->header_fn(snp_fh, &hdr);
bam_sspileup_file(fp1, fp2, d->mask, d->mapQ, glf_somatic, d, snp_fh);
}
else {
fprintf(stderr, "Unable to open snp file!!!!!!!!!\n");
exit(1);
}
bam_close(fp1);
bam_close(fp2);
bam_header_destroy(d->h1);
bam_header_destroy(d->h2);
if (d->fai) fai_destroy(d->fai);
sniper_maqcns_destroy(d->c);
free(d->ref); free(d);
fclose(snp_fh);
return 0;
}
/*!
@abstract Sort an unsorted BAM file based on the chromosome order
and the leftmost position of an alignment
@param is_by_qname whether to sort by query name
@param fn name of the file to be sorted
@param prefix prefix of the output and the temporary files; upon
sucessess, prefix.bam will be written.
@param max_mem approxiate maximum memory (very inaccurate)
@discussion It may create multiple temporary subalignment files
and then merge them by calling bam_merge_core(). This function is
NOT thread safe.
*/
void bam_sort_core_ext(int is_by_qname, const char *fn, const char *prefix, size_t max_mem, int is_stdout)
{
int n, ret, k, i;
size_t mem;
bam_header_t *header;
bamFile fp;
bam1_t *b, **buf;
g_is_by_qname = is_by_qname;
n = k = 0; mem = 0;
fp = strcmp(fn, "-")? bam_open(fn, "r") : bam_dopen(fileno(stdin), "r");
if (fp == 0) {
fprintf(stderr, "[bam_sort_core] fail to open file %s\n", fn);
return;
}
header = bam_header_read(fp);
if (is_by_qname) change_SO(header, "queryname");
else change_SO(header, "coordinate");
buf = (bam1_t**)calloc(max_mem / BAM_CORE_SIZE, sizeof(bam1_t*));
// write sub files
for (;;) {
if (buf[k] == 0) buf[k] = (bam1_t*)calloc(1, sizeof(bam1_t));
b = buf[k];
if ((ret = bam_read1(fp, b)) < 0) break;
mem += ret;
++k;
if (mem >= max_mem) {
sort_blocks(n++, k, buf, prefix, header, 0);
mem = 0; k = 0;
}
}
if (ret != -1)
fprintf(stderr, "[bam_sort_core] truncated file. Continue anyway.\n");
if (n == 0) sort_blocks(-1, k, buf, prefix, header, is_stdout);
else { // then merge
char **fns, *fnout;
fprintf(stderr, "[bam_sort_core] merging from %d files...\n", n+1);
sort_blocks(n++, k, buf, prefix, header, 0);
fnout = (char*)calloc(strlen(prefix) + 20, 1);
if (is_stdout) sprintf(fnout, "-");
else sprintf(fnout, "%s.bam", prefix);
fns = (char**)calloc(n, sizeof(char*));
for (i = 0; i < n; ++i) {
fns[i] = (char*)calloc(strlen(prefix) + 20, 1);
sprintf(fns[i], "%s.%.4d.bam", prefix, i);
}
bam_merge_core(is_by_qname, fnout, 0, n, fns, 0, 0);
free(fnout);
for (i = 0; i < n; ++i) {
unlink(fns[i]);
free(fns[i]);
}
free(fns);
}
for (k = 0; (size_t)k < max_mem / BAM_CORE_SIZE; ++k) {
if (buf[k]) {
free(buf[k]->data);
free(buf[k]);
}
}
free(buf);
bam_header_destroy(header);
bam_close(fp);
}
samfile_t *samopen(const char *fn, const char *mode, const void *aux)
{
samfile_t *fp;
fp = (samfile_t*)calloc(1, sizeof(samfile_t));
if (mode[0] == 'r') { // read
fp->type |= TYPE_READ;
if (mode[1] == 'b') { // binary
fp->type |= TYPE_BAM;
fp->x.bam = strcmp(fn, "-")? bam_open(fn, "r") : bam_dopen(fileno(stdin), "r");
if (fp->x.bam == 0) goto open_err_ret;
fp->header = bam_header_read(fp->x.bam);
} else { // text
fp->x.tamr = sam_open(fn);
if (fp->x.tamr == 0) goto open_err_ret;
fp->header = sam_header_read(fp->x.tamr);
if (fp->header->n_targets == 0) { // no @SQ fields
if (aux) { // check if aux is present
bam_header_destroy(fp->header);
fp->header = sam_header_read2((const char*)aux);
}
if (fp->header->n_targets == 0)
fprintf(stderr, "[samopen] empty header.\n");
} else fprintf(stderr, "[samopen] SAM header is present: %d sequences.\n", fp->header->n_targets);
}
} else if (mode[0] == 'w') { // write
fp->header = bam_header_dup((const bam_header_t*)aux);
if (mode[1] == 'b') { // binary
fp->type |= TYPE_BAM;
fp->x.bam = strcmp(fn, "-")? bam_open(fn, "w") : bam_dopen(fileno(stdout), "w");
if (fp->x.bam == 0) goto open_err_ret;
bam_header_write(fp->x.bam, fp->header);
} else { // text
// open file
fp->x.tamw = strcmp(fn, "-")? fopen(fn, "w") : stdout;
if (fp->x.tamr == 0) goto open_err_ret;
// write header
if (strstr(mode, "h")) {
int i;
bam_header_t *alt;
// parse the header text
alt = bam_header_init();
alt->l_text = fp->header->l_text; alt->text = fp->header->text;
sam_header_parse(alt);
alt->l_text = 0; alt->text = 0;
// check if there are @SQ lines in the header
if (alt->n_targets) { // then write the header text without dumping ->target_{name,len}
if (alt->n_targets != fp->header->n_targets)
fprintf(stderr, "[samopen] inconsistent number of target sequences.\n");
fwrite(fp->header->text, 1, fp->header->l_text, fp->x.tamw);
} else { // then dump ->target_{name,len}
for (i = 0; i < fp->header->n_targets; ++i)
fprintf(fp->x.tamw, "@SQ\tSN:%s\tLN:%d\n", fp->header->target_name[i], fp->header->target_len[i]);
}
bam_header_destroy(alt);
}
}
}
return fp;
open_err_ret:
free(fp);
return 0;
}
static int mpileup(mplp_conf_t *conf, int n, char **fn)
{
extern void *bcf_call_add_rg(void *rghash, const char *hdtext, const char *list);
extern void bcf_call_del_rghash(void *rghash);
mplp_aux_t **data;
int i, tid, pos, *n_plp, tid0 = -1, beg0 = 0, end0 = 1u<<29, ref_len, ref_tid = -1, max_depth, max_indel_depth;
const bam_pileup1_t **plp;
bam_mplp_t iter;
bam_header_t *h = 0;
char *ref;
void *rghash = 0;
bcf_callaux_t *bca = 0;
bcf_callret1_t *bcr = 0;
bcf_call_t bc;
bcf_t *bp = 0;
bcf_hdr_t *bh = 0;
bam_sample_t *sm = 0;
kstring_t buf;
mplp_pileup_t gplp;
memset(&gplp, 0, sizeof(mplp_pileup_t));
memset(&buf, 0, sizeof(kstring_t));
memset(&bc, 0, sizeof(bcf_call_t));
data = calloc(n, sizeof(void*));
plp = calloc(n, sizeof(void*));
n_plp = calloc(n, sizeof(int*));
sm = bam_smpl_init();
// read the header and initialize data
for (i = 0; i < n; ++i) {
bam_header_t *h_tmp;
data[i] = calloc(1, sizeof(mplp_aux_t));
data[i]->fp = strcmp(fn[i], "-") == 0? bam_dopen(fileno(stdin), "r") : bam_open(fn[i], "r");
data[i]->conf = conf;
h_tmp = bam_header_read(data[i]->fp);
data[i]->h = i? h : h_tmp; // for i==0, "h" has not been set yet
bam_smpl_add(sm, fn[i], (conf->flag&MPLP_IGNORE_RG)? 0 : h_tmp->text);
rghash = bcf_call_add_rg(rghash, h_tmp->text, conf->pl_list);
if (conf->reg) {
int beg, end;
bam_index_t *idx;
idx = bam_index_load(fn[i]);
if (idx == 0) {
fprintf(stderr, "[%s] fail to load index for %d-th input.\n", __func__, i+1);
exit(1);
}
if (bam_parse_region(h_tmp, conf->reg, &tid, &beg, &end) < 0) {
fprintf(stderr, "[%s] malformatted region or wrong seqname for %d-th input.\n", __func__, i+1);
exit(1);
}
if (i == 0) tid0 = tid, beg0 = beg, end0 = end;
data[i]->iter = bam_iter_query(idx, tid, beg, end);
bam_index_destroy(idx);
}
if (i == 0) h = h_tmp;
else {
// FIXME: to check consistency
bam_header_destroy(h_tmp);
}
}
gplp.n = sm->n;
gplp.n_plp = calloc(sm->n, sizeof(int));
gplp.m_plp = calloc(sm->n, sizeof(int));
gplp.plp = calloc(sm->n, sizeof(void*));
fprintf(stderr, "[%s] %d samples in %d input files\n", __func__, sm->n, n);
// write the VCF header
if (conf->flag & MPLP_GLF) {
kstring_t s;
bh = calloc(1, sizeof(bcf_hdr_t));
s.l = s.m = 0; s.s = 0;
bp = bcf_open("-", (conf->flag&MPLP_NO_COMP)? "wu" : "w");
for (i = 0; i < h->n_targets; ++i) {
kputs(h->target_name[i], &s);
kputc('\0', &s);
}
bh->l_nm = s.l;
bh->name = malloc(s.l);
memcpy(bh->name, s.s, s.l);
s.l = 0;
for (i = 0; i < sm->n; ++i) {
kputs(sm->smpl[i], &s); kputc('\0', &s);
}
bh->l_smpl = s.l;
bh->sname = malloc(s.l);
memcpy(bh->sname, s.s, s.l);
bh->txt = malloc(strlen(BAM_VERSION) + 64);
bh->l_txt = 1 + sprintf(bh->txt, "##samtoolsVersion=%s\n", BAM_VERSION);
free(s.s);
bcf_hdr_sync(bh);
bcf_hdr_write(bp, bh);
bca = bcf_call_init(-1., conf->min_baseQ);
bcr = calloc(sm->n, sizeof(bcf_callret1_t));
bca->rghash = rghash;
bca->openQ = conf->openQ, bca->extQ = conf->extQ, bca->tandemQ = conf->tandemQ;
bca->min_frac = conf->min_frac;
bca->min_support = conf->min_support;
}
if (tid0 >= 0 && conf->fai) { // region is set
ref = faidx_fetch_seq(conf->fai, h->target_name[tid0], 0, 0x7fffffff, &ref_len);
ref_tid = tid0;
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid0;
} else ref_tid = -1, ref = 0;
iter = bam_mplp_init(n, mplp_func, (void**)data);
max_depth = conf->max_depth;
if (max_depth * sm->n > 1<<20)
fprintf(stderr, "(%s) Max depth is above 1M. Potential memory hog!\n", __func__);
if (max_depth * sm->n < 8000) {
max_depth = 8000 / sm->n;
fprintf(stderr, "<%s> Set max per-file depth to %d\n", __func__, max_depth);
}
max_indel_depth = conf->max_indel_depth * sm->n;
bam_mplp_set_maxcnt(iter, max_depth);
int storeSize = 100;
int delStore[2][100] = {{0},{0}};
typedef char * mstring;
while (bam_mplp_auto(iter, &tid, &pos, n_plp, plp) > 0) {
if (conf->reg && (pos < beg0 || pos >= end0)) continue; // out of the region requested
if (conf->bed && tid >= 0 && !bed_overlap(conf->bed, h->target_name[tid], pos, pos+1)) continue;
if (tid != ref_tid) {
free(ref); ref = 0;
if (conf->fai) ref = faidx_fetch_seq(conf->fai, h->target_name[tid], 0, 0x7fffffff, &ref_len);
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid;
ref_tid = tid;
}
if (conf->flag & MPLP_GLF) {
int total_depth, _ref0, ref16;
bcf1_t *b = calloc(1, sizeof(bcf1_t));
for (i = total_depth = 0; i < n; ++i) total_depth += n_plp[i];
group_smpl(&gplp, sm, &buf, n, fn, n_plp, plp, conf->flag & MPLP_IGNORE_RG);
_ref0 = (ref && pos < ref_len)? ref[pos] : 'N';
ref16 = bam_nt16_table[_ref0];
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], ref16, bca, bcr + i);
bcf_call_combine(gplp.n, bcr, ref16, &bc);
bcf_call2bcf(tid, pos, &bc, b, (conf->flag&(MPLP_FMT_DP|MPLP_FMT_SP))? bcr : 0,
(conf->flag&MPLP_FMT_SP), 0, 0);
bcf_write(bp, bh, b);
bcf_destroy(b);
// call indels
if (!(conf->flag&MPLP_NO_INDEL) && total_depth < max_indel_depth && bcf_call_gap_prep(gplp.n, gplp.n_plp, gplp.plp, pos, bca, ref, rghash) >= 0) {
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], -1, bca, bcr + i);
if (bcf_call_combine(gplp.n, bcr, -1, &bc) >= 0) {
b = calloc(1, sizeof(bcf1_t));
bcf_call2bcf(tid, pos, &bc, b, (conf->flag&(MPLP_FMT_DP|MPLP_FMT_SP))? bcr : 0,
(conf->flag&MPLP_FMT_SP), bca, ref);
bcf_write(bp, bh, b);
bcf_destroy(b);
}
}
} else {
printf("%s\t%d\t%c", h->target_name[tid], pos + 1, (ref && pos < ref_len)? ref[pos] : 'N');
for (i = 0; i < n; ++i) {
int j;
printf("\t%d\t", n_plp[i]);
if (n_plp[i] == 0) {
printf("*\t*"); // FIXME: printf() is very slow...
if (conf->flag & MPLP_PRINT_POS) printf("\t*");
} else {
//MDW start
//for each position in the pileup column
int charLen = 16;
int countChars[ charLen ][2];
int countiChars[ charLen ][2];
int countGap[2]={0,0};
//double qvTotal=0;
int numStruck=0;
int numGood=0;
int tti;
int ttj;
mstring insAllele[100];
int insAlleleCnt[100];
int sf=0;
int flag=0;
//typedef char * string;
char insStr0[10000];
int iCnt0=0;
char insStr1[10000];
int iCnt1=0;
char delStr0[10000];
int dCnt0=0;
char delStr1[10000];
int dCnt1=0;
float qposP[10000];
int qposCnt=0;
//initialize with zeros
for(tti=0;tti<charLen;tti++){
countChars[tti][0]=0;
countChars[tti][1]=0;
}
// define repeat length here; look back up to 10 prior positions
// start one position away.
int replC=0; //
for(tti=1;tti<=15;tti++){
// check for greater than zero
if(toupper(ref[pos-1])==toupper(ref[pos-tti])){
replC++;
}else{ // breaks the chain at first non identical to current position not strict homopolymer
break;
}
}
int reprC=0; //
for(tti=1;tti<=15;tti++){
// check for greater than zero
if(toupper(ref[pos+1])==toupper(ref[pos+tti])){
reprC++;
}else{ // breaks the chain at first non identical to current position not strict homopolymer
break;
}
}
int repT = replC;
if(replC < reprC){
repT=reprC;
}
for (j = 0; j < n_plp[i]; ++j){
const bam_pileup1_t *p = plp[i] + j;
/*
SAME LOGIC AS pileup_seq()
*/
if(p->is_refskip){ // never count intron gaps in numStruck
continue;
}
if(p->is_del){ // skip deletion gap, after first position which is the first aligned char
continue;
}
if( p->b->core.qual < conf->min_mqToCount || // mapping quality
conf->maxrepC < (repT) || // max homopolymer run, this will not
(!p->is_del && bam1_qual(p->b)[p->qpos] < conf->min_baseQ) || // base quality for matches
p->alignedQPosBeg <= (conf->trimEnd ) || p->alignedQPosEnd <= (conf->trimEnd ) || // trimEnd is 1-based
p->zf == 1 || // fusion tag
p->ih > conf->maxIH || // max hit index
(p->nmd > conf->maxNM) || // max mismatch
(conf->flagFilter == 1 && !(p->b->core.flag&BAM_FPROPER_PAIR)) || // optionally keep only proper pairs
(conf->flagFilter == 2 && p->b->core.flag&BAM_FSECONDARY) || // optionally strike secondary
(conf->flagFilter == 3 && p->b->core.flag&BAM_FDUP) || // optionally strike dup
(conf->flagFilter == 4 && (p->b->core.flag&BAM_FDUP || p->b->core.flag&BAM_FSECONDARY)) || // optionally strike secondary or dup
(conf->flagFilter == 5 && (p->b->core.flag&BAM_FDUP || p->b->core.flag&BAM_FSECONDARY || p->b->core.flag&BAM_FQCFAIL || !(p->b->core.flag&BAM_FPROPER_PAIR) )) // optionally strike secondary, dup and QCfail
){
numStruck++;
continue;
}
//printf("repT=%d: %d %c %c %c %c \n",repT,p->indel,ref[pos],ref[pos-1],ref[pos-2],ref[pos-3]);
if(!p->is_del && p->indel==0){
countChars[ bam1_seqi(bam1_seq(p->b), p->qpos) ][ bam1_strand(p->b) ] ++;
numGood++;
}else if(p->is_refskip){
countGap[ bam1_strand(p->b) ]++;
}
if(p->indel<0){
numGood++;
if(bam1_strand(p->b) ==0){
for(tti=1;tti<= -p->indel; tti++) {
// current spot, starting at 0 in store, because indel<0 refers to next position
delStr0[dCnt0] = ref[pos+tti];
dCnt0++;
}
delStr0[dCnt0] = ',';
dCnt0++;
}else{
for(tti=1;tti<= -p->indel; tti++) {
// current spot, starting at 0 in store, because indel<0 refers to next position
delStr1[dCnt1] = ref[pos+tti];
dCnt1++;
}
delStr1[dCnt1] = ',';
dCnt1++;
}
}else if(p->indel>0){
numGood++;
if(bam1_strand(p->b) ==0){
for(tti=1;tti<= p->indel; tti++) {
// current spot, starting at 0 in store, because indel<0 refers to next position
insStr0[iCnt0] = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos + tti)];
iCnt0++;
}
insStr0[iCnt0] = ',';
iCnt0++;
}else{
for(tti=1;tti<= p->indel; tti++) {
// current spot, starting at 0 in store, because indel<0 refers to next position
insStr1[iCnt1] = bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos + tti)];
iCnt1++;
}
insStr1[iCnt1] = ',';
iCnt1++;
}
}
//calculate position of variant within aligned read - no soft clips
if( toupper(ref[pos]) != toupper(bam_nt16_rev_table[bam1_seqi(bam1_seq(p->b), p->qpos)]) || p->indel>0 || p->indel<0 ){
//distance to end; calculate distance to end of aligned read. removes soft clips.
int distToEnd = (p->alignedQPosBeg < p->alignedQPosEnd) ? p->alignedQPosBeg : p->alignedQPosEnd;
qposP[qposCnt] = distToEnd;
qposCnt++;
// printf("id=%s, pos=%d",bam1_qname(p->b),distToEnd);
}
}
//
//print A,C,G,T, by +/-
printf("\t%d\t%d\t%d\t%d\t%d\t%d\t%d\t%d\t%d\t%d", countChars[1][0],countChars[1][1],
countChars[2][0],countChars[2][1],
countChars[4][0],countChars[4][1],
countChars[8][0],countChars[8][1],
countChars[7][0],countChars[7][1]);
putchar('\t');
for(tti=0;tti<dCnt0;tti++){
putchar(delStr0[tti]);
}
putchar('\t');
for(tti=0;tti<dCnt1;tti++){
putchar(delStr1[tti]);
}
putchar('\t');
for(tti=0;tti<iCnt0;tti++){
putchar(insStr0[tti]);
}
putchar('\t');
for(tti=0;tti<iCnt1;tti++){
putchar(insStr1[tti]);
}
printf("\t%d\t%d",numGood,numStruck);
// get non-ref qpos variation
float medqpos = -1;
float medAbsDev = -1;
if(qposCnt>0){
medqpos = median(qposCnt,qposP);
float absDev[qposCnt];
for(tti=0;tti<qposCnt;tti++){
absDev[tti] = abs(medqpos - qposP[tti]);
}
medAbsDev = median(qposCnt-1,absDev);
}
printf("\t%f",medAbsDev);
///END MDW
}
}
putchar('\n');
}
}
bcf_close(bp);
bam_smpl_destroy(sm); free(buf.s);
for (i = 0; i < gplp.n; ++i) free(gplp.plp[i]);
free(gplp.plp); free(gplp.n_plp); free(gplp.m_plp);
bcf_call_del_rghash(rghash);
bcf_hdr_destroy(bh); bcf_call_destroy(bca); free(bc.PL); free(bcr);
bam_mplp_destroy(iter);
bam_header_destroy(h);
for (i = 0; i < n; ++i) {
bam_close(data[i]->fp);
if (data[i]->iter) bam_iter_destroy(data[i]->iter);
free(data[i]);
}
free(data); free(plp); free(ref); free(n_plp);
return 0;
}
int bam_merge_core2(int by_qname, const char *out, const char *headers, int n, char * const *fn, int flag, const char *reg, int level)
#endif
{
bamFile fpout, *fp;
heap1_t *heap;
bam_header_t *hout = 0;
bam_header_t *hheaders = NULL;
int i, j, *RG_len = 0;
uint64_t idx = 0;
char **RG = 0, mode[8];
bam_iter_t *iter = 0;
if (headers) {
tamFile fpheaders = sam_open(headers);
if (fpheaders == 0) {
const char *message = strerror(errno);
fprintf(stderr, "[bam_merge_core] cannot open '%s': %s\n", headers, message);
return -1;
}
hheaders = sam_header_read(fpheaders);
sam_close(fpheaders);
}
g_is_by_qname = by_qname;
fp = (bamFile*)calloc(n, sizeof(bamFile));
heap = (heap1_t*)calloc(n, sizeof(heap1_t));
iter = (bam_iter_t*)calloc(n, sizeof(bam_iter_t));
// prepare RG tag
if (flag & MERGE_RG) {
RG = (char**)calloc(n, sizeof(void*));
RG_len = (int*)calloc(n, sizeof(int));
for (i = 0; i != n; ++i) {
int l = strlen(fn[i]);
const char *s = fn[i];
if (l > 4 && strcmp(s + l - 4, ".bam") == 0) l -= 4;
for (j = l - 1; j >= 0; --j) if (s[j] == '/') break;
++j; l -= j;
RG[i] = calloc(l + 1, 1);
RG_len[i] = l;
strncpy(RG[i], s + j, l);
}
}
// read the first
for (i = 0; i != n; ++i) {
bam_header_t *hin;
fp[i] = bam_open(fn[i], "r");
if (fp[i] == 0) {
int j;
fprintf(stderr, "[bam_merge_core] fail to open file %s\n", fn[i]);
for (j = 0; j < i; ++j) bam_close(fp[j]);
free(fp); free(heap);
// FIXME: possible memory leak
return -1;
}
hin = bam_header_read(fp[i]);
if (i == 0) { // the first BAM
hout = hin;
} else { // validate multiple baf
int min_n_targets = hout->n_targets;
if (hin->n_targets < min_n_targets) min_n_targets = hin->n_targets;
for (j = 0; j < min_n_targets; ++j)
if (strcmp(hout->target_name[j], hin->target_name[j]) != 0) {
fprintf(stderr, "[bam_merge_core] different target sequence name: '%s' != '%s' in file '%s'\n",
hout->target_name[j], hin->target_name[j], fn[i]);
return -1;
}
// If this input file has additional target reference sequences,
// add them to the headers to be output
if (hin->n_targets > hout->n_targets) {
swap_header_targets(hout, hin);
// FIXME Possibly we should also create @SQ text headers
// for the newly added reference sequences
}
bam_header_destroy(hin);
}
}
if (hheaders) {
// If the text headers to be swapped in include any @SQ headers,
// check that they are consistent with the existing binary list
// of reference information.
if (hheaders->n_targets > 0) {
if (hout->n_targets != hheaders->n_targets) {
fprintf(stderr, "[bam_merge_core] number of @SQ headers in '%s' differs from number of target sequences\n", headers);
if (!reg) return -1;
}
for (j = 0; j < hout->n_targets; ++j)
if (strcmp(hout->target_name[j], hheaders->target_name[j]) != 0) {
fprintf(stderr, "[bam_merge_core] @SQ header '%s' in '%s' differs from target sequence\n", hheaders->target_name[j], headers);
if (!reg) return -1;
}
}
swap_header_text(hout, hheaders);
bam_header_destroy(hheaders);
}
if (reg) {
int tid, beg, end;
if (bam_parse_region(hout, reg, &tid, &beg, &end) < 0) {
fprintf(stderr, "[%s] Malformated region string or undefined reference name\n", __func__);
return -1;
}
for (i = 0; i < n; ++i) {
bam_index_t *idx;
idx = bam_index_load(fn[i]);
iter[i] = bam_iter_query(idx, tid, beg, end);
bam_index_destroy(idx);
}
}
for (i = 0; i < n; ++i) {
heap1_t *h = heap + i;
h->i = i;
h->b = (bam1_t*)calloc(1, sizeof(bam1_t));
if (bam_iter_read(fp[i], iter[i], h->b) >= 0) {
h->pos = ((uint64_t)h->b->core.tid<<32) | (uint32_t)((int32_t)h->b->core.pos+1)<<1 | bam1_strand(h->b);
h->idx = idx++;
}
else h->pos = HEAP_EMPTY;
}
if (flag & MERGE_UNCOMP) level = 0;
else if (flag & MERGE_LEVEL1) level = 1;
strcpy(mode, "w");
if (level >= 0) sprintf(mode + 1, "%d", level < 9? level : 9);
if ((fpout = strcmp(out, "-")? bam_open(out, "w") : bam_dopen(fileno(stdout), "w")) == 0) {
fprintf(stderr, "[%s] fail to create the output file.\n", __func__);
return -1;
}
bam_header_write(fpout, hout);
bam_header_destroy(hout);
#ifndef _PBGZF_USE
if (!(flag & MERGE_UNCOMP)) bgzf_mt(fpout, n_threads, 256);
#endif
ks_heapmake(heap, n, heap);
while (heap->pos != HEAP_EMPTY) {
bam1_t *b = heap->b;
if (flag & MERGE_RG) {
uint8_t *rg = bam_aux_get(b, "RG");
if (rg) bam_aux_del(b, rg);
bam_aux_append(b, "RG", 'Z', RG_len[heap->i] + 1, (uint8_t*)RG[heap->i]);
}
bam_write1_core(fpout, &b->core, b->data_len, b->data);
if ((j = bam_iter_read(fp[heap->i], iter[heap->i], b)) >= 0) {
heap->pos = ((uint64_t)b->core.tid<<32) | (uint32_t)((int)b->core.pos+1)<<1 | bam1_strand(b);
heap->idx = idx++;
} else if (j == -1) {
heap->pos = HEAP_EMPTY;
free(heap->b->data); free(heap->b);
heap->b = 0;
} else fprintf(stderr, "[bam_merge_core] '%s' is truncated. Continue anyway.\n", fn[heap->i]);
ks_heapadjust(heap, 0, n, heap);
}
if (flag & MERGE_RG) {
for (i = 0; i != n; ++i) free(RG[i]);
free(RG); free(RG_len);
}
for (i = 0; i != n; ++i) {
bam_iter_destroy(iter[i]);
bam_close(fp[i]);
}
bam_close(fpout);
free(fp); free(heap); free(iter);
return 0;
}
/*!
@abstract Sort an unsorted BAM file based on the chromosome order
and the leftmost position of an alignment
@param is_by_qname whether to sort by query name
@param fn name of the file to be sorted
@param prefix prefix of the output and the temporary files; upon
sucessess, prefix.bam will be written.
@param max_mem approxiate maximum memory (very inaccurate)
@discussion It may create multiple temporary subalignment files
and then merge them by calling bam_merge_core(). This function is
NOT thread safe.
*/
void bam_sort_core_ext(int is_by_qname, const char *fn, const char *prefix, size_t _max_mem, int is_stdout, int n_threads, int level, int sort_type)
{
int ret, i, n_files = 0;
size_t mem, max_k, k, max_mem;
bam_header_t *header;
bamFile fp;
bam1_t *b, **buf;
char *fnout = 0;
if (n_threads < 2) n_threads = 1;
g_is_by_qname = is_by_qname;
max_k = k = 0; mem = 0;
max_mem = _max_mem * n_threads;
buf = 0;
fp = strcmp(fn, "-")? bam_open(fn, "r") : bam_dopen(fileno(stdin), "r");
if (fp == 0) {
fprintf(stderr, "[bam_sort_core] fail to open file %s\n", fn);
return;
}
header = bam_header_read(fp);
if (is_by_qname) change_SO(header, "queryname");
else change_SO(header, "coordinate");
// write sub files
for (;;) {
if (k == max_k) {
size_t old_max = max_k;
max_k = max_k? max_k<<1 : 0x10000;
buf = realloc(buf, max_k * sizeof(void*));
memset(buf + old_max, 0, sizeof(void*) * (max_k - old_max));
}
if (buf[k] == 0) buf[k] = (bam1_t*)calloc(1, sizeof(bam1_t));
b = buf[k];
if ((ret = bam_read1(fp, b)) < 0) break;
if (b->data_len < b->m_data>>2) { // shrink
b->m_data = b->data_len;
kroundup32(b->m_data);
b->data = realloc(b->data, b->m_data);
}
mem += sizeof(bam1_t) + b->m_data + sizeof(void*) + sizeof(void*); // two sizeof(void*) for the data allocated to pointer arrays
++k;
if (mem >= max_mem) {
n_files = sort_blocks(n_files, k, buf, prefix, header, n_threads, sort_type);
mem = k = 0;
}
}
if (ret != -1)
fprintf(stderr, "[bam_sort_core] truncated file. Continue anyway.\n");
// output file name
fnout = calloc(strlen(prefix) + 20, 1);
if (is_stdout) sprintf(fnout, "-");
else sprintf(fnout, "%s.bam", prefix);
// write the final output
if (n_files == 0) { // a single block
char mode[8];
strcpy(mode, "w");
if (level >= 0) sprintf(mode + 1, "%d", level < 9? level : 9);
sort_aux_core(k, buf, sort_type);
#ifndef _PBGZF_USE
write_buffer(fnout, mode, k, buf, header, n_threads);
#else
write_buffer(fnout, mode, k, buf, header);
#endif
} else { // then merge
char **fns;
n_files = sort_blocks(n_files, k, buf, prefix, header, n_threads, sort_type);
fprintf(stderr, "[bam_sort_core] merging from %d files...\n", n_files);
fns = (char**)calloc(n_files, sizeof(char*));
for (i = 0; i < n_files; ++i) {
fns[i] = (char*)calloc(strlen(prefix) + 20, 1);
sprintf(fns[i], "%s.%.4d.bam", prefix, i);
}
#ifndef _PBGZF_USE
bam_merge_core2(is_by_qname, fnout, 0, n_files, fns, 0, 0, n_threads, level);
#else
bam_merge_core2(is_by_qname, fnout, 0, n_files, fns, 0, 0, level);
#endif
for (i = 0; i < n_files; ++i) {
unlink(fns[i]);
free(fns[i]);
}
free(fns);
}
free(fnout);
// free
for (k = 0; k < max_k; ++k) {
if (!buf[k]) continue;
free(buf[k]->data);
free(buf[k]);
}
free(buf);
bam_header_destroy(header);
bam_close(fp);
}
void bam_header_free(bam_header_t *header) {
bam_header_destroy(header);
}
samfile_t *samopen(const char *fn, const char *mode, const void *aux)
{
samfile_t *fp;
fp = (samfile_t*)calloc(1, sizeof(samfile_t));
if (strchr(mode, 'r')) { // read
fp->type |= TYPE_READ;
if (strchr(mode, 'b')) { // binary
fp->type |= TYPE_BAM;
fp->x.bam = strcmp(fn, "-")? bam_open(fn, "r") : bam_dopen(fileno(stdin), "r");
if (fp->x.bam == 0) goto open_err_ret;
fp->header = bam_header_read(fp->x.bam);
} else { // text
fp->x.tamr = sam_open(fn);
if (fp->x.tamr == 0) goto open_err_ret;
fp->header = sam_header_read(fp->x.tamr);
if (fp->header->n_targets == 0) { // no @SQ fields
if (aux) { // check if aux is present
bam_header_t *textheader = fp->header;
fp->header = sam_header_read2((const char*)aux);
if (fp->header == 0) goto open_err_ret;
append_header_text(fp->header, textheader->text, textheader->l_text);
bam_header_destroy(textheader);
}
if (fp->header->n_targets == 0 && bam_verbose >= 1)
fprintf(stderr, "[samopen] no @SQ lines in the header.\n");
} //else if (bam_verbose >= 2) fprintf(stderr, "[samopen] SAM header is present: %d sequences.\n", fp->header->n_targets);
}
} else if (strchr(mode, 'w')) { // write
fp->header = bam_header_dup((const bam_header_t*)aux);
if (strchr(mode, 'b')) { // binary
char bmode[3];
int i, compress_level = -1;
for (i = 0; mode[i]; ++i) if (mode[i] >= '0' && mode[i] <= '9') break;
if (mode[i]) compress_level = mode[i] - '0';
if (strchr(mode, 'u')) compress_level = 0;
bmode[0] = 'w'; bmode[1] = compress_level < 0? 0 : compress_level + '0'; bmode[2] = 0;
fp->type |= TYPE_BAM;
fp->x.bam = strcmp(fn, "-")? bam_open(fn, bmode) : bam_dopen(fileno(stdout), bmode);
if (fp->x.bam == 0) goto open_err_ret;
bam_header_write(fp->x.bam, fp->header);
} else { // text
// open file
fp->x.tamw = strcmp(fn, "-")? fopen(fn, "w") : stdout;
if (fp->x.tamw == 0) goto open_err_ret;
if (strchr(mode, 'X')) fp->type |= BAM_OFSTR<<2;
else if (strchr(mode, 'x')) fp->type |= BAM_OFHEX<<2;
else fp->type |= BAM_OFDEC<<2;
// write header
if (strchr(mode, 'h')) {
int i;
bam_header_t *alt;
// parse the header text
alt = bam_header_init();
alt->l_text = fp->header->l_text; alt->text = fp->header->text;
sam_header_parse(alt);
alt->l_text = 0; alt->text = 0;
// check if there are @SQ lines in the header
fwrite(fp->header->text, 1, fp->header->l_text, fp->x.tamw); // FIXME: better to skip the trailing NULL
if (alt->n_targets) { // then write the header text without dumping ->target_{name,len}
if (alt->n_targets != fp->header->n_targets && bam_verbose >= 1)
fprintf(stderr, "[samopen] inconsistent number of target sequences. Output the text header.\n");
} else { // then dump ->target_{name,len}
for (i = 0; i < fp->header->n_targets; ++i)
fprintf(fp->x.tamw, "@SQ\tSN:%s\tLN:%d\n", fp->header->target_name[i], fp->header->target_len[i]);
}
bam_header_destroy(alt);
}
}
}
return fp;
open_err_ret:
free(fp);
return 0;
}
static int mpileup(mplp_conf_t *conf, int n, char **fn)
{
extern void *bcf_call_add_rg(void *rghash, const char *hdtext, const char *list);
extern void bcf_call_del_rghash(void *rghash);
mplp_aux_t **data;
int i, tid, pos, *n_plp, tid0 = -1, beg0 = 0, end0 = 1u<<29, ref_len, ref_tid = -1, max_depth, max_indel_depth;
const bam_pileup1_t **plp;
bam_mplp_t iter;
bam_header_t *h = 0;
char *ref;
void *rghash = 0;
bcf_callaux_t *bca = 0;
bcf_callret1_t *bcr = 0;
bcf_call_t bc;
bcf_t *bp = 0;
bcf_hdr_t *bh = 0;
bam_sample_t *sm = 0;
kstring_t buf;
mplp_pileup_t gplp;
memset(&gplp, 0, sizeof(mplp_pileup_t));
memset(&buf, 0, sizeof(kstring_t));
memset(&bc, 0, sizeof(bcf_call_t));
data = calloc(n, sizeof(void*));
plp = calloc(n, sizeof(void*));
n_plp = calloc(n, sizeof(int*));
sm = bam_smpl_init();
// read the header and initialize data
for (i = 0; i < n; ++i) {
bam_header_t *h_tmp;
data[i] = calloc(1, sizeof(mplp_aux_t));
data[i]->fp = strcmp(fn[i], "-") == 0? bam_dopen(fileno(stdin), "r") : bam_open(fn[i], "r");
data[i]->conf = conf;
h_tmp = bam_header_read(data[i]->fp);
data[i]->h = i? h : h_tmp; // for i==0, "h" has not been set yet
bam_smpl_add(sm, fn[i], (conf->flag&MPLP_IGNORE_RG)? 0 : h_tmp->text);
rghash = bcf_call_add_rg(rghash, h_tmp->text, conf->pl_list);
if (conf->reg) {
int beg, end;
bam_index_t *idx;
idx = bam_index_load(fn[i]);
if (idx == 0) {
fprintf(stderr, "[%s] fail to load index for %d-th input.\n", __func__, i+1);
exit(1);
}
if (bam_parse_region(h_tmp, conf->reg, &tid, &beg, &end) < 0) {
fprintf(stderr, "[%s] malformatted region or wrong seqname for %d-th input.\n", __func__, i+1);
exit(1);
}
if (i == 0) tid0 = tid, beg0 = beg, end0 = end;
data[i]->iter = bam_iter_query(idx, tid, beg, end);
bam_index_destroy(idx);
}
if (i == 0) h = h_tmp;
else {
// FIXME: to check consistency
bam_header_destroy(h_tmp);
}
}
gplp.n = sm->n;
gplp.n_plp = calloc(sm->n, sizeof(int));
gplp.m_plp = calloc(sm->n, sizeof(int));
gplp.plp = calloc(sm->n, sizeof(void*));
fprintf(stderr, "[%s] %d samples in %d input files\n", __func__, sm->n, n);
// write the VCF header
if (conf->flag & MPLP_GLF) {
kstring_t s;
bh = calloc(1, sizeof(bcf_hdr_t));
s.l = s.m = 0; s.s = 0;
bp = bcf_open("-", (conf->flag&MPLP_NO_COMP)? "wu" : "w");
for (i = 0; i < h->n_targets; ++i) {
kputs(h->target_name[i], &s);
kputc('\0', &s);
}
bh->l_nm = s.l;
bh->name = malloc(s.l);
memcpy(bh->name, s.s, s.l);
s.l = 0;
for (i = 0; i < sm->n; ++i) {
kputs(sm->smpl[i], &s); kputc('\0', &s);
}
bh->l_smpl = s.l;
bh->sname = malloc(s.l);
memcpy(bh->sname, s.s, s.l);
bh->txt = malloc(strlen(BAM_VERSION) + 64);
bh->l_txt = 1 + sprintf(bh->txt, "##samtoolsVersion=%s\n", BAM_VERSION);
free(s.s);
bcf_hdr_sync(bh);
bcf_hdr_write(bp, bh);
bca = bcf_call_init(-1., conf->min_baseQ);
bcr = calloc(sm->n, sizeof(bcf_callret1_t));
bca->rghash = rghash;
bca->openQ = conf->openQ, bca->extQ = conf->extQ, bca->tandemQ = conf->tandemQ;
bca->min_frac = conf->min_frac;
bca->min_support = conf->min_support;
}
if (tid0 >= 0 && conf->fai) { // region is set
ref = faidx_fetch_seq(conf->fai, h->target_name[tid0], 0, 0x7fffffff, &ref_len);
ref_tid = tid0;
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid0;
} else ref_tid = -1, ref = 0;
iter = bam_mplp_init(n, mplp_func, (void**)data);
max_depth = conf->max_depth;
if (max_depth * sm->n > 1<<20)
fprintf(stderr, "(%s) Max depth is above 1M. Potential memory hog!\n", __func__);
if (max_depth * sm->n < 8000) {
max_depth = 8000 / sm->n;
fprintf(stderr, "<%s> Set max per-file depth to %d\n", __func__, max_depth);
}
max_indel_depth = conf->max_indel_depth * sm->n;
bam_mplp_set_maxcnt(iter, max_depth);
while (bam_mplp_auto(iter, &tid, &pos, n_plp, plp) > 0) {
if (conf->reg && (pos < beg0 || pos >= end0)) continue; // out of the region requested
if (conf->bed && tid >= 0 && !bed_overlap(conf->bed, h->target_name[tid], pos, pos+1)) continue;
if (tid != ref_tid) {
free(ref); ref = 0;
if (conf->fai) ref = faidx_fetch_seq(conf->fai, h->target_name[tid], 0, 0x7fffffff, &ref_len);
for (i = 0; i < n; ++i) data[i]->ref = ref, data[i]->ref_id = tid;
ref_tid = tid;
}
if (conf->flag & MPLP_GLF) {
int total_depth, _ref0, ref16;
bcf1_t *b = calloc(1, sizeof(bcf1_t));
for (i = total_depth = 0; i < n; ++i) total_depth += n_plp[i];
group_smpl(&gplp, sm, &buf, n, fn, n_plp, plp, conf->flag & MPLP_IGNORE_RG);
_ref0 = (ref && pos < ref_len)? ref[pos] : 'N';
ref16 = bam_nt16_table[_ref0];
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], ref16, bca, bcr + i);
bcf_call_combine(gplp.n, bcr, ref16, &bc);
bcf_call2bcf(tid, pos, &bc, b, (conf->flag&(MPLP_FMT_DP|MPLP_FMT_SP))? bcr : 0,
(conf->flag&MPLP_FMT_SP), 0, 0);
bcf_write(bp, bh, b);
bcf_destroy(b);
// call indels
if (!(conf->flag&MPLP_NO_INDEL) && total_depth < max_indel_depth && bcf_call_gap_prep(gplp.n, gplp.n_plp, gplp.plp, pos, bca, ref, rghash) >= 0) {
for (i = 0; i < gplp.n; ++i)
bcf_call_glfgen(gplp.n_plp[i], gplp.plp[i], -1, bca, bcr + i);
if (bcf_call_combine(gplp.n, bcr, -1, &bc) >= 0) {
b = calloc(1, sizeof(bcf1_t));
bcf_call2bcf(tid, pos, &bc, b, (conf->flag&(MPLP_FMT_DP|MPLP_FMT_SP))? bcr : 0,
(conf->flag&MPLP_FMT_SP), bca, ref);
bcf_write(bp, bh, b);
bcf_destroy(b);
}
}
} else {
printf("%s\t%d\t%c", h->target_name[tid], pos + 1, (ref && pos < ref_len)? ref[pos] : 'N');
for (i = 0; i < n; ++i) {
int j;
printf("\t%d\t", n_plp[i]);
if (n_plp[i] == 0) {
printf("*\t*"); // FIXME: printf() is very slow...
if (conf->flag & MPLP_PRINT_POS) printf("\t*");
} else {
for (j = 0; j < n_plp[i]; ++j)
pileup_seq(plp[i] + j, pos, ref_len, ref);
putchar('\t');
for (j = 0; j < n_plp[i]; ++j) {
const bam_pileup1_t *p = plp[i] + j;
int c = bam1_qual(p->b)[p->qpos] + 33;
if (c > 126) c = 126;
putchar(c);
}
if (conf->flag & MPLP_PRINT_MAPQ) {
putchar('\t');
for (j = 0; j < n_plp[i]; ++j) {
int c = plp[i][j].b->core.qual + 33;
if (c > 126) c = 126;
putchar(c);
}
}
if (conf->flag & MPLP_PRINT_POS) {
putchar('\t');
for (j = 0; j < n_plp[i]; ++j) {
if (j > 0) putchar(',');
printf("%d", plp[i][j].qpos + 1); // FIXME: printf() is very slow...
}
}
}
}
putchar('\n');
}
}
bcf_close(bp);
bam_smpl_destroy(sm); free(buf.s);
for (i = 0; i < gplp.n; ++i) free(gplp.plp[i]);
free(gplp.plp); free(gplp.n_plp); free(gplp.m_plp);
bcf_call_del_rghash(rghash);
bcf_hdr_destroy(bh); bcf_call_destroy(bca); free(bc.PL); free(bcr);
bam_mplp_destroy(iter);
bam_header_destroy(h);
for (i = 0; i < n; ++i) {
bam_close(data[i]->fp);
if (data[i]->iter) bam_iter_destroy(data[i]->iter);
free(data[i]);
}
free(data); free(plp); free(ref); free(n_plp);
return 0;
}
int main_bedcov(int argc, char *argv[])
{
extern void bam_init_header_hash(bam_header_t*);
gzFile fp;
kstring_t str;
kstream_t *ks;
bam_index_t **idx;
bam_header_t *h = 0;
aux_t **aux;
int *n_plp, dret, i, n, c, min_mapQ = 0;
int64_t *cnt;
const bam_pileup1_t **plp;
while ((c = getopt(argc, argv, "Q:")) >= 0) {
switch (c) {
case 'Q': min_mapQ = atoi(optarg); break;
}
}
if (optind + 2 > argc) {
fprintf(stderr, "Usage: samtools bedcov <in.bed> <in1.bam> [...]\n");
return 1;
}
memset(&str, 0, sizeof(kstring_t));
n = argc - optind - 1;
aux = calloc(n, sizeof(aux_t*));
idx = calloc(n, sizeof(bam_index_t*));
for (i = 0; i < n; ++i) {
aux[i] = calloc(1, sizeof(aux_t));
aux[i]->min_mapQ = min_mapQ;
aux[i]->fp = bam_open(argv[i+optind+1], "r");
idx[i] = bam_index_load(argv[i+optind+1]);
if (aux[i]->fp == 0 || idx[i] == 0) {
fprintf(stderr, "ERROR: fail to open index BAM file '%s'\n", argv[i+optind+1]);
return 2;
}
bgzf_set_cache_size(aux[i]->fp, 20);
if (i == 0) h = bam_header_read(aux[0]->fp);
}
bam_init_header_hash(h);
cnt = calloc(n, 8);
fp = gzopen(argv[optind], "rb");
ks = ks_init(fp);
n_plp = calloc(n, sizeof(int));
plp = calloc(n, sizeof(bam_pileup1_t*));
while (ks_getuntil(ks, KS_SEP_LINE, &str, &dret) >= 0) {
char *p, *q;
int tid, beg, end, pos;
bam_mplp_t mplp;
for (p = q = str.s; *p && *p != '\t'; ++p);
if (*p != '\t') goto bed_error;
*p = 0; tid = bam_get_tid(h, q); *p = '\t';
if (tid < 0) goto bed_error;
for (q = p = p + 1; isdigit(*p); ++p);
if (*p != '\t') goto bed_error;
*p = 0; beg = atoi(q); *p = '\t';
for (q = p = p + 1; isdigit(*p); ++p);
if (*p == '\t' || *p == 0) {
int c = *p;
*p = 0; end = atoi(q); *p = c;
} else goto bed_error;
for (i = 0; i < n; ++i) {
if (aux[i]->iter) bam_iter_destroy(aux[i]->iter);
aux[i]->iter = bam_iter_query(idx[i], tid, beg, end);
}
mplp = bam_mplp_init(n, read_bam, (void**)aux);
bam_mplp_set_maxcnt(mplp, 64000);
memset(cnt, 0, 8 * n);
while (bam_mplp_auto(mplp, &tid, &pos, n_plp, plp) > 0)
if (pos >= beg && pos < end)
for (i = 0; i < n; ++i) cnt[i] += n_plp[i];
for (i = 0; i < n; ++i) {
kputc('\t', &str);
kputl(cnt[i], &str);
}
puts(str.s);
bam_mplp_destroy(mplp);
continue;
bed_error:
fprintf(stderr, "Errors in BED line '%s'\n", str.s);
}
free(n_plp); free(plp);
ks_destroy(ks);
gzclose(fp);
free(cnt);
for (i = 0; i < n; ++i) {
if (aux[i]->iter) bam_iter_destroy(aux[i]->iter);
bam_index_destroy(idx[i]);
bam_close(aux[i]->fp);
free(aux[i]);
}
bam_header_destroy(h);
free(aux); free(idx);
free(str.s);
return 0;
}
