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#include "pysam.h"
#include <stdlib.h>
#include <string.h>
#include <stdio.h>
#include <unistd.h>
#include <math.h>
#include <inttypes.h>
#include "sam_header.h"
#include "sam.h"
#include "faidx.h"
#include "kstring.h"
#include "khash.h"
KHASH_SET_INIT_STR(rg)
// When counting records instead of printing them,
// data passed to the bam_fetch callback is encapsulated in this struct.
typedef struct {
bam_header_t *header;
int64_t *count; // int does overflow for very big BAMs
} count_func_data_t;
typedef khash_t(rg) *rghash_t;
// FIXME: we'd better use no global variables...
static rghash_t g_rghash = 0;
static int g_min_mapQ = 0, g_flag_on = 0, g_flag_off = 0, g_qual_scale = 0, g_min_qlen = 0;
static uint32_t g_subsam_seed = 0;
static double g_subsam_frac = -1.;
static char *g_library, *g_rg;
static void *g_bed;
void *bed_read(const char *fn);
void bed_destroy(void *_h);
int bed_overlap(const void *_h, const char *chr, int beg, int end);
static int process_aln(const bam_header_t *h, bam1_t *b)
{
if (g_qual_scale > 1) {
int i;
uint8_t *qual = bam1_qual(b);
for (i = 0; i < b->core.l_qseq; ++i) {
int c = qual[i] * g_qual_scale;
qual[i] = c < 93? c : 93;
}
}
if (g_min_qlen > 0) {
int k, qlen = 0;
uint32_t *cigar = bam1_cigar(b);
for (k = 0; k < b->core.n_cigar; ++k)
if ((bam_cigar_type(bam_cigar_op(cigar[k]))&1) || bam_cigar_op(cigar[k]) == BAM_CHARD_CLIP)
qlen += bam_cigar_oplen(cigar[k]);
if (qlen < g_min_qlen) return 1;
}
if (b->core.qual < g_min_mapQ || ((b->core.flag & g_flag_on) != g_flag_on) || (b->core.flag & g_flag_off))
return 1;
if (g_bed && b->core.tid >= 0 && !bed_overlap(g_bed, h->target_name[b->core.tid], b->core.pos, bam_calend(&b->core, bam1_cigar(b))))
return 1;
if (g_subsam_frac > 0.) {
uint32_t k = __ac_X31_hash_string(bam1_qname(b)) + g_subsam_seed;
if ((double)(k&0xffffff) / 0x1000000 >= g_subsam_frac) return 1;
}
if (g_rg || g_rghash) {
uint8_t *s = bam_aux_get(b, "RG");
if (s) {
if (g_rg) return (strcmp(g_rg, (char*)(s + 1)) == 0)? 0 : 1;
if (g_rghash) {
khint_t k = kh_get(rg, g_rghash, (char*)(s + 1));
return (k != kh_end(g_rghash))? 0 : 1;
}
}
}
if (g_library) {
const char *p = bam_get_library((bam_header_t*)h, b);
return (p && strcmp(p, g_library) == 0)? 0 : 1;
}
return 0;
}
static char *drop_rg(char *hdtxt, rghash_t h, int *len)
{
char *p = hdtxt, *q, *r, *s;
kstring_t str;
memset(&str, 0, sizeof(kstring_t));
while (1) {
int toprint = 0;
q = strchr(p, '\n');
if (q == 0) q = p + strlen(p);
if (q - p < 3) break; // the line is too short; then stop
if (strncmp(p, "@RG\t", 4) == 0) {
int c;
khint_t k;
if ((r = strstr(p, "\tID:")) != 0) {
r += 4;
for (s = r; *s != '\0' && *s != '\n' && *s != '\t'; ++s);
c = *s; *s = '\0';
k = kh_get(rg, h, r);
*s = c;
if (k != kh_end(h)) toprint = 1;
}
} else toprint = 1;
if (toprint) {
kputsn(p, q - p, &str); kputc('\n', &str);
}
p = q + 1;
}
*len = str.l;
return str.s;
}
// callback function for bam_fetch() that prints nonskipped records
static int view_func(const bam1_t *b, void *data)
{
if (!process_aln(((samfile_t*)data)->header, (bam1_t*)b))
samwrite((samfile_t*)data, b);
return 0;
}
// callback function for bam_fetch() that counts nonskipped records
static int count_func(const bam1_t *b, void *data)
{
if (!process_aln(((count_func_data_t*)data)->header, (bam1_t*)b)) {
(*((count_func_data_t*)data)->count)++;
}
return 0;
}
static int usage(int is_long_help);
int main_samview(int argc, char *argv[])
{
int c, is_header = 0, is_header_only = 0, is_bamin = 1, ret = 0, compress_level = -1, is_bamout = 0, is_count = 0;
int of_type = BAM_OFDEC, is_long_help = 0, n_threads = 0;
int64_t count = 0;
samfile_t *in = 0, *out = 0;
char in_mode[5], out_mode[5], *fn_out = 0, *fn_list = 0, *fn_ref = 0, *fn_rg = 0, *q;
/* parse command-line options */
strcpy(in_mode, "r"); strcpy(out_mode, "w");
while ((c = getopt(argc, argv, "SbBct:h1Ho:q:f:F:ul:r:xX?T:R:L:s:Q:@:m:")) >= 0) {
switch (c) {
case 's':
if ((g_subsam_seed = strtol(optarg, &q, 10)) != 0) {
srand(g_subsam_seed);
g_subsam_seed = rand();
}
g_subsam_frac = strtod(q, &q);
break;
case 'm': g_min_qlen = atoi(optarg); break;
case 'c': is_count = 1; break;
case 'S': is_bamin = 0; break;
case 'b': is_bamout = 1; break;
case 't': fn_list = strdup(optarg); is_bamin = 0; break;
case 'h': is_header = 1; break;
case 'H': is_header_only = 1; break;
case 'o': fn_out = strdup(optarg); break;
case 'f': g_flag_on = strtol(optarg, 0, 0); break;
case 'F': g_flag_off = strtol(optarg, 0, 0); break;
case 'q': g_min_mapQ = atoi(optarg); break;
case 'u': compress_level = 0; break;
case '1': compress_level = 1; break;
case 'l': g_library = strdup(optarg); break;
case 'L': g_bed = bed_read(optarg); break;
case 'r': g_rg = strdup(optarg); break;
case 'R': fn_rg = strdup(optarg); break;
case 'x': of_type = BAM_OFHEX; break;
case 'X': of_type = BAM_OFSTR; break;
case '?': is_long_help = 1; break;
case 'T': fn_ref = strdup(optarg); is_bamin = 0; break;
case 'B': bam_no_B = 1; break;
case 'Q': g_qual_scale = atoi(optarg); break;
case '@': n_threads = strtol(optarg, 0, 0); break;
default: return usage(is_long_help);
}
}
if (compress_level >= 0) is_bamout = 1;
if (is_header_only) is_header = 1;
if (is_bamout) strcat(out_mode, "b");
else {
if (of_type == BAM_OFHEX) strcat(out_mode, "x");
else if (of_type == BAM_OFSTR) strcat(out_mode, "X");
}
if (is_bamin) strcat(in_mode, "b");
if (is_header) strcat(out_mode, "h");
if (compress_level >= 0) {
char tmp[2];
tmp[0] = compress_level + '0'; tmp[1] = '\0';
strcat(out_mode, tmp);
}
if (argc == optind) return usage(is_long_help); // potential memory leak...
// read the list of read groups
if (fn_rg) {
FILE *fp_rg;
char buf[1024];
int ret;
g_rghash = kh_init(rg);
fp_rg = fopen(fn_rg, "r");
while (!feof(fp_rg) && fscanf(fp_rg, "%s", buf) > 0) // this is not a good style, but bear me...
kh_put(rg, g_rghash, strdup(buf), &ret); // we'd better check duplicates...
fclose(fp_rg);
}
// generate the fn_list if necessary
if (fn_list == 0 && fn_ref) fn_list = samfaipath(fn_ref);
// open file handlers
if ((in = samopen(argv[optind], in_mode, fn_list)) == 0) {
fprintf(pysamerr, "[main_samview] fail to open \"%s\" for reading.\n", argv[optind]);
ret = 1;
goto view_end;
}
if (in->header == 0) {
fprintf(pysamerr, "[main_samview] fail to read the header from \"%s\".\n", argv[optind]);
ret = 1;
goto view_end;
}
if (g_rghash) { // FIXME: I do not know what "bam_header_t::n_text" is for...
char *tmp;
int l;
tmp = drop_rg(in->header->text, g_rghash, &l);
free(in->header->text);
in->header->text = tmp;
in->header->l_text = l;
}
if (!is_count && (out = samopen(fn_out? fn_out : "-", out_mode, in->header)) == 0) {
fprintf(pysamerr, "[main_samview] fail to open \"%s\" for writing.\n", fn_out? fn_out : "standard output");
ret = 1;
goto view_end;
}
if (n_threads > 1) samthreads(out, n_threads, 256);
if (is_header_only) goto view_end; // no need to print alignments
if (argc == optind + 1) { // convert/print the entire file
bam1_t *b = bam_init1();
int r;
while ((r = samread(in, b)) >= 0) { // read one alignment from `in'
if (!process_aln(in->header, b)) {
if (!is_count) samwrite(out, b); // write the alignment to `out'
count++;
}
}
if (r < -1) {
fprintf(pysamerr, "[main_samview] truncated file.\n");
ret = 1;
}
bam_destroy1(b);
} else { // retrieve alignments in specified regions
int i;
bam_index_t *idx = 0;
if (is_bamin) idx = bam_index_load(argv[optind]); // load BAM index
if (idx == 0) { // index is unavailable
fprintf(pysamerr, "[main_samview] random alignment retrieval only works for indexed BAM files.\n");
ret = 1;
goto view_end;
}
for (i = optind + 1; i < argc; ++i) {
int tid, beg, end, result;
bam_parse_region(in->header, argv[i], &tid, &beg, &end); // parse a region in the format like `chr2:100-200'
if (tid < 0) { // reference name is not found
fprintf(pysamerr, "[main_samview] region \"%s\" specifies an unknown reference name. Continue anyway.\n", argv[i]);
continue;
}
// fetch alignments
if (is_count) {
count_func_data_t count_data = { in->header, &count };
result = bam_fetch(in->x.bam, idx, tid, beg, end, &count_data, count_func);
} else
result = bam_fetch(in->x.bam, idx, tid, beg, end, out, view_func);
if (result < 0) {
fprintf(pysamerr, "[main_samview] retrieval of region \"%s\" failed due to truncated file or corrupt BAM index file\n", argv[i]);
ret = 1;
break;
}
}
bam_index_destroy(idx); // destroy the BAM index
}
view_end:
if (is_count && ret == 0)
printf("%" PRId64 "\n", count);
// close files, free and return
free(fn_list); free(fn_ref); free(fn_out); free(g_library); free(g_rg); free(fn_rg);
if (g_bed) bed_destroy(g_bed);
if (g_rghash) {
khint_t k;
for (k = 0; k < kh_end(g_rghash); ++k)
if (kh_exist(g_rghash, k)) free((char*)kh_key(g_rghash, k));
kh_destroy(rg, g_rghash);
}
samclose(in);
if (!is_count)
samclose(out);
return ret;
}
static int usage(int is_long_help)
{
fprintf(pysamerr, "\n");
fprintf(pysamerr, "Usage: samtools view [options] <in.bam>|<in.sam> [region1 [...]]\n\n");
fprintf(pysamerr, "Options: -b output BAM\n");
fprintf(pysamerr, " -h print header for the SAM output\n");
fprintf(pysamerr, " -H print header only (no alignments)\n");
fprintf(pysamerr, " -S input is SAM\n");
fprintf(pysamerr, " -u uncompressed BAM output (force -b)\n");
fprintf(pysamerr, " -1 fast compression (force -b)\n");
fprintf(pysamerr, " -x output FLAG in HEX (samtools-C specific)\n");
fprintf(pysamerr, " -X output FLAG in string (samtools-C specific)\n");
fprintf(pysamerr, " -c print only the count of matching records\n");
fprintf(pysamerr, " -B collapse the backward CIGAR operation\n");
fprintf(pysamerr, " -@ INT number of BAM compression threads [0]\n");
fprintf(pysamerr, " -L FILE output alignments overlapping the input BED FILE [null]\n");
fprintf(pysamerr, " -t FILE list of reference names and lengths (force -S) [null]\n");
fprintf(pysamerr, " -T FILE reference sequence file (force -S) [null]\n");
fprintf(pysamerr, " -o FILE output file name [stdout]\n");
fprintf(pysamerr, " -R FILE list of read groups to be outputted [null]\n");
fprintf(pysamerr, " -f INT required flag, 0 for unset [0]\n");
fprintf(pysamerr, " -F INT filtering flag, 0 for unset [0]\n");
fprintf(pysamerr, " -q INT minimum mapping quality [0]\n");
fprintf(pysamerr, " -l STR only output reads in library STR [null]\n");
fprintf(pysamerr, " -r STR only output reads in read group STR [null]\n");
fprintf(pysamerr, " -s FLOAT fraction of templates to subsample; integer part as seed [-1]\n");
fprintf(pysamerr, " -? longer help\n");
fprintf(pysamerr, "\n");
if (is_long_help)
fprintf(pysamerr, "Notes:\n\
\n\
1. By default, this command assumes the file on the command line is in\n\
the BAM format and it prints the alignments in SAM. If `-t' is\n\
applied, the input file is assumed to be in the SAM format. The\n\
file supplied with `-t' is SPACE/TAB delimited with the first two\n\
fields of each line consisting of the reference name and the\n\
corresponding sequence length. The `.fai' file generated by `faidx'\n\
can be used here. This file may be empty if reads are unaligned.\n\
\n\
2. SAM->BAM conversion: `samtools view -bT ref.fa in.sam.gz'.\n\
\n\
3. BAM->SAM conversion: `samtools view in.bam'.\n\
\n\
4. A region should be presented in one of the following formats:\n\
`chr1', `chr2:1,000' and `chr3:1000-2,000'. When a region is\n\
specified, the input alignment file must be an indexed BAM file.\n\
\n\
5. Option `-u' is preferred over `-b' when the output is piped to\n\
another samtools command.\n\
\n\
6. In a string FLAG, each character represents one bit with\n\
p=0x1 (paired), P=0x2 (properly paired), u=0x4 (unmapped),\n\
U=0x8 (mate unmapped), r=0x10 (reverse), R=0x20 (mate reverse)\n\
1=0x40 (first), 2=0x80 (second), s=0x100 (not primary), \n\
f=0x200 (failure) and d=0x400 (duplicate). Note that `-x' and\n\
`-X' are samtools-C specific. Picard and older samtools do not\n\
support HEX or string flags.\n\
\n");
return 1;
}
int main_import(int argc, char *argv[])
{
int argc2, ret;
char **argv2;
if (argc != 4) {
fprintf(pysamerr, "Usage: bamtk import <in.ref_list> <in.sam> <out.bam>\n");
return 1;
}
argc2 = 6;
argv2 = calloc(6, sizeof(char*));
argv2[0] = "import", argv2[1] = "-o", argv2[2] = argv[3], argv2[3] = "-bt", argv2[4] = argv[1], argv2[5] = argv[2];
ret = main_samview(argc2, argv2);
free(argv2);
return ret;
}
int8_t seq_comp_table[16] = { 0, 8, 4, 12, 2, 10, 9, 14, 1, 6, 5, 13, 3, 11, 7, 15 };
int main_bam2fq(int argc, char *argv[])
{
bamFile fp;
bam_header_t *h;
bam1_t *b;
int8_t *buf;
int max_buf, c, no12 = 0;
while ((c = getopt(argc, argv, "n")) > 0)
if (c == 'n') no12 = 1;
if (argc == 1) {
fprintf(pysamerr, "Usage: samtools bam2fq <in.bam>\n");
return 1;
}
fp = strcmp(argv[optind], "-")? bam_open(argv[optind], "r") : bam_dopen(fileno(stdin), "r");
if (fp == 0) return 1;
h = bam_header_read(fp);
b = bam_init1();
buf = 0;
max_buf = 0;
while (bam_read1(fp, b) >= 0) {
int i, qlen = b->core.l_qseq;
uint8_t *seq;
putchar('@'); fputs(bam1_qname(b), stdout);
if (no12) putchar('\n');
else {
if ((b->core.flag & 0x40) && !(b->core.flag & 0x80)) puts("/1");
else if ((b->core.flag & 0x80) && !(b->core.flag & 0x40)) puts("/2");
else putchar('\n');
}
if (max_buf < qlen + 1) {
max_buf = qlen + 1;
kroundup32(max_buf);
buf = realloc(buf, max_buf);
}
buf[qlen] = 0;
seq = bam1_seq(b);
for (i = 0; i < qlen; ++i)
buf[i] = bam1_seqi(seq, i);
if (b->core.flag & 16) { // reverse complement
for (i = 0; i < qlen>>1; ++i) {
int8_t t = seq_comp_table[buf[qlen - 1 - i]];
buf[qlen - 1 - i] = seq_comp_table[buf[i]];
buf[i] = t;
}
if (qlen&1) buf[i] = seq_comp_table[buf[i]];
}
for (i = 0; i < qlen; ++i)
buf[i] = bam_nt16_rev_table[buf[i]];
puts((char*)buf);
puts("+");
seq = bam1_qual(b);
for (i = 0; i < qlen; ++i)
buf[i] = 33 + seq[i];
if (b->core.flag & 16) { // reverse
for (i = 0; i < qlen>>1; ++i) {
int8_t t = buf[qlen - 1 - i];
buf[qlen - 1 - i] = buf[i];
buf[i] = t;
}
}
puts((char*)buf);
}
free(buf);
bam_destroy1(b);
bam_header_destroy(h);
bam_close(fp);
return 0;
}
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