void into_menu()
{
char imagename[16];
switch(main_menu_point)
{
case 0: work = BOOK_MENU;
workdircase = ONLY_FILE;
strcpy(workdirname, "book");
strcpy(imagename, "sys/book.api");
break;
case 1: work = MUSIC_MENU;
workdircase = ONLY_FILE;
strcpy(workdirname, "/");
strcpy(imagename, "sys/music.api");
break;
case 2: work = IMAGE_MENU;
workdircase = ONLY_FILE;
strcpy(workdirname, "image");
strcpy(imagename, "sys/image.api");
break;
case 3: work = GAME_MENU;
workdircase = ONLY_DIR;
strcpy(workdirname, "game");
strcpy(imagename, "sys/game.api");
break;
case 4: work = FILE_MENU;
workdircase = BOTH_FILE_DIR;
strcpy(workdirname, "/");
strcpy(imagename, "sys/file.api");
break;
case 5: into_about();
return;
}
myGLCD.fillScr(255, 255, 255);
show_apimg(23, 23, imagename); //显示图标
file_amount = get_file_amount(workdirname, workdircase); //文件数量
file_offset = 0;
read_file_list(workdirname, file_offset, workdircase); //载入文件列表
if(file_amount)
{
show_file_menu(); //显示文件列表
file_list_point = 0;
myGLCD.setColor(0, 0, 255);
myGLCD.drawRect(210, 2, 218, 173);
draw_file_list_point(file_list_point, 255, 0, 0); //光标
}
else
show_chinese_sentence(111, 4, "\xCE\xDE\xCE\xC4\xBC\xFE", 0, 0, 0, 1); //无文件
return;
}
int bam_mpileup(int argc, char *argv[])
{
int c;
const char *file_list = NULL;
char **fn = NULL;
int nfiles = 0, use_orphan = 0;
mplp_conf_t mplp;
memset(&mplp, 0, sizeof(mplp_conf_t));
#define MPLP_PRINT_POS 0x4000
mplp.max_mq = 60;
mplp.min_baseQ = 13;
mplp.capQ_thres = 0;
mplp.max_depth = 250; mplp.max_indel_depth = 250;
mplp.openQ = 40; mplp.extQ = 20; mplp.tandemQ = 100;
mplp.min_frac = 0.002; mplp.min_support = 1;
mplp.flag = MPLP_NO_ORPHAN | MPLP_REALN;
while ((c = getopt(argc, argv, "Agf:r:l:M:q:Q:uaRC:BDSd:L:b:P:o:e:h:Im:F:EG:6Os")) >= 0) {
switch (c) {
case 'f':
mplp.fai = fai_load(optarg);
if (mplp.fai == 0) return 1;
break;
case 'd': mplp.max_depth = atoi(optarg); break;
case 'r': mplp.reg = strdup(optarg); break;
case 'l': mplp.bed = bed_read(optarg); break;
case 'P': mplp.pl_list = strdup(optarg); break;
case 'g': mplp.flag |= MPLP_GLF; break;
case 'u': mplp.flag |= MPLP_NO_COMP | MPLP_GLF; break;
case 'a': mplp.flag |= MPLP_NO_ORPHAN | MPLP_REALN; break;
case 'B': mplp.flag &= ~MPLP_REALN; break;
case 'D': mplp.flag |= MPLP_FMT_DP; break;
case 'S': mplp.flag |= MPLP_FMT_SP; break;
case 'I': mplp.flag |= MPLP_NO_INDEL; break;
case 'E': mplp.flag |= MPLP_EXT_BAQ; break;
case '6': mplp.flag |= MPLP_ILLUMINA13; break;
case 'R': mplp.flag |= MPLP_IGNORE_RG; break;
case 's': mplp.flag |= MPLP_PRINT_MAPQ; break;
case 'O': mplp.flag |= MPLP_PRINT_POS; break;
case 'C': mplp.capQ_thres = atoi(optarg); break;
case 'M': mplp.max_mq = atoi(optarg); break;
case 'q': mplp.min_mq = atoi(optarg); break;
case 'Q': mplp.min_baseQ = atoi(optarg); break;
case 'b': file_list = optarg; break;
case 'o': mplp.openQ = atoi(optarg); break;
case 'e': mplp.extQ = atoi(optarg); break;
case 'h': mplp.tandemQ = atoi(optarg); break;
case 'A': use_orphan = 1; break;
case 'F': mplp.min_frac = atof(optarg); break;
case 'm': mplp.min_support = atoi(optarg); break;
case 'L': mplp.max_indel_depth = atoi(optarg); break;
case 'G': {
FILE *fp_rg;
char buf[1024];
mplp.rghash = bcf_str2id_init();
if ((fp_rg = fopen(optarg, "r")) == 0)
fprintf(stderr, "(%s) Fail to open file %s. Continue anyway.\n", __func__, optarg);
while (!feof(fp_rg) && fscanf(fp_rg, "%s", buf) > 0) // this is not a good style, but forgive me...
bcf_str2id_add(mplp.rghash, strdup(buf));
fclose(fp_rg);
}
break;
}
}
if (use_orphan) mplp.flag &= ~MPLP_NO_ORPHAN;
if (argc == 1) {
fprintf(stderr, "\n");
fprintf(stderr, "Usage: samtools mpileup [options] in1.bam [in2.bam [...]]\n\n");
fprintf(stderr, "Input options:\n\n");
fprintf(stderr, " -6 assume the quality is in the Illumina-1.3+ encoding\n");
fprintf(stderr, " -A count anomalous read pairs\n");
fprintf(stderr, " -B disable BAQ computation\n");
fprintf(stderr, " -b FILE list of input BAM files [null]\n");
fprintf(stderr, " -C INT parameter for adjusting mapQ; 0 to disable [0]\n");
fprintf(stderr, " -d INT max per-BAM depth to avoid excessive memory usage [%d]\n", mplp.max_depth);
fprintf(stderr, " -E extended BAQ for higher sensitivity but lower specificity\n");
fprintf(stderr, " -f FILE faidx indexed reference sequence file [null]\n");
fprintf(stderr, " -G FILE exclude read groups listed in FILE [null]\n");
fprintf(stderr, " -l FILE list of positions (chr pos) or regions (BED) [null]\n");
fprintf(stderr, " -M INT cap mapping quality at INT [%d]\n", mplp.max_mq);
fprintf(stderr, " -r STR region in which pileup is generated [null]\n");
fprintf(stderr, " -R ignore RG tags\n");
fprintf(stderr, " -q INT skip alignments with mapQ smaller than INT [%d]\n", mplp.min_mq);
fprintf(stderr, " -Q INT skip bases with baseQ/BAQ smaller than INT [%d]\n", mplp.min_baseQ);
fprintf(stderr, "\nOutput options:\n\n");
fprintf(stderr, " -D output per-sample DP in BCF (require -g/-u)\n");
fprintf(stderr, " -g generate BCF output (genotype likelihoods)\n");
fprintf(stderr, " -O output base positions on reads (disabled by -g/-u)\n");
fprintf(stderr, " -s output mapping quality (disabled by -g/-u)\n");
fprintf(stderr, " -S output per-sample strand bias P-value in BCF (require -g/-u)\n");
fprintf(stderr, " -u generate uncompress BCF output\n");
fprintf(stderr, "\nSNP/INDEL genotype likelihoods options (effective with `-g' or `-u'):\n\n");
fprintf(stderr, " -e INT Phred-scaled gap extension seq error probability [%d]\n", mplp.extQ);
fprintf(stderr, " -F FLOAT minimum fraction of gapped reads for candidates [%g]\n", mplp.min_frac);
fprintf(stderr, " -h INT coefficient for homopolymer errors [%d]\n", mplp.tandemQ);
fprintf(stderr, " -I do not perform indel calling\n");
fprintf(stderr, " -L INT max per-sample depth for INDEL calling [%d]\n", mplp.max_indel_depth);
fprintf(stderr, " -m INT minimum gapped reads for indel candidates [%d]\n", mplp.min_support);
fprintf(stderr, " -o INT Phred-scaled gap open sequencing error probability [%d]\n", mplp.openQ);
fprintf(stderr, " -P STR comma separated list of platforms for indels [all]\n");
fprintf(stderr, "\n");
fprintf(stderr, "Notes: Assuming diploid individuals.\n\n");
return 1;
}
if (file_list) {
if ( read_file_list(file_list,&nfiles,&fn) ) return 1;
mpileup(&mplp,nfiles,fn);
for (c=0; c<nfiles; c++) free(fn[c]);
free(fn);
} else mpileup(&mplp, argc - optind, argv + optind);
if (mplp.rghash) bcf_str2id_thorough_destroy(mplp.rghash);
free(mplp.reg); free(mplp.pl_list);
if (mplp.fai) fai_destroy(mplp.fai);
if (mplp.bed) bed_destroy(mplp.bed);
return 0;
}
int main_depth(int argc, char *argv[])
{
int i, n, tid, reg_tid, beg, end, pos, *n_plp, baseQ = 0, mapQ = 0, min_len = 0;
int all = 0, status = EXIT_SUCCESS, nfiles, max_depth = -1;
const bam_pileup1_t **plp;
char *reg = 0; // specified region
void *bed = 0; // BED data structure
char *file_list = NULL, **fn = NULL;
bam_hdr_t *h = NULL; // BAM header of the 1st input
aux_t **data;
bam_mplp_t mplp;
int last_pos = -1, last_tid = -1, ret;
sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
static const struct option lopts[] = {
SAM_OPT_GLOBAL_OPTIONS('-', 0, '-', '-', 0),
{ NULL, 0, NULL, 0 }
};
// parse the command line
while ((n = getopt_long(argc, argv, "r:b:q:Q:l:f:am:d:", lopts, NULL)) >= 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); // BED or position list file can be parsed now
if (!bed) {
print_error_errno("depth", "Could not read file \"%s\"", optarg);
return 1;
}
break;
case 'q':
baseQ = atoi(optarg);
break; // base quality threshold
case 'Q':
mapQ = atoi(optarg);
break; // mapping quality threshold
case 'f':
file_list = optarg;
break;
case 'a':
all++;
break;
case 'd':
case 'm':
max_depth = atoi(optarg);
break; // maximum coverage depth
default:
if (parse_sam_global_opt(n, optarg, lopts, &ga) == 0) break;
/* else fall-through */
case '?':
return usage();
}
}
if (optind == argc && !file_list)
return usage();
// 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(aux_t*)); // data[i] for the i-th input
reg_tid = 0;
beg = 0;
end = INT_MAX; // set the default region
for (i = 0; i < n; ++i) {
int rf;
data[i] = calloc(1, sizeof(aux_t));
data[i]->fp = sam_open_format(argv[optind+i], "r", &ga.in); // open BAM
if (data[i]->fp == NULL) {
print_error_errno("depth", "Could not open \"%s\"", argv[optind+i]);
status = EXIT_FAILURE;
goto depth_end;
}
rf = SAM_FLAG | SAM_RNAME | SAM_POS | SAM_MAPQ | SAM_CIGAR | SAM_SEQ;
if (baseQ) rf |= SAM_QUAL;
if (hts_set_opt(data[i]->fp, CRAM_OPT_REQUIRED_FIELDS, rf)) {
fprintf(stderr, "Failed to set CRAM_OPT_REQUIRED_FIELDS value\n");
return 1;
}
if (hts_set_opt(data[i]->fp, CRAM_OPT_DECODE_MD, 0)) {
fprintf(stderr, "Failed to set CRAM_OPT_DECODE_MD value\n");
return 1;
}
data[i]->min_mapQ = mapQ; // set the mapQ filter
data[i]->min_len = min_len; // set the qlen filter
data[i]->hdr = sam_hdr_read(data[i]->fp); // read the BAM header
if (data[i]->hdr == NULL) {
fprintf(stderr, "Couldn't read header for \"%s\"\n",
argv[optind+i]);
status = EXIT_FAILURE;
goto depth_end;
}
if (reg) { // if a region is specified
hts_idx_t *idx = sam_index_load(data[i]->fp, argv[optind+i]); // load the index
if (idx == NULL) {
print_error("depth", "can't load index for \"%s\"", argv[optind+i]);
status = EXIT_FAILURE;
goto depth_end;
}
data[i]->iter = sam_itr_querys(idx, data[i]->hdr, reg); // set the iterator
hts_idx_destroy(idx); // the index is not needed any more; free the memory
if (data[i]->iter == NULL) {
print_error("depth", "can't parse region \"%s\"", reg);
status = EXIT_FAILURE;
goto depth_end;
}
}
}
h = data[0]->hdr; // easy access to the header of the 1st BAM
if (reg) {
beg = data[0]->iter->beg; // and to the parsed region coordinates
end = data[0]->iter->end;
reg_tid = data[0]->iter->tid;
}
// the core multi-pileup loop
mplp = bam_mplp_init(n, read_bam, (void**)data); // initialization
if (0 < max_depth)
bam_mplp_set_maxcnt(mplp,max_depth); // set maximum coverage depth
n_plp = calloc(n, sizeof(int)); // n_plp[i] is the number of covering reads from the i-th BAM
plp = calloc(n, sizeof(bam_pileup1_t*)); // plp[i] points to the array of covering reads (internal in mplp)
while ((ret=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 (tid >= h->n_targets) continue; // diff number of @SQ lines per file?
if (all) {
while (tid > last_tid) {
if (last_tid >= 0 && !reg) {
// Deal with remainder or entirety of last tid.
while (++last_pos < h->target_len[last_tid]) {
// Horribly inefficient, but the bed API is an obfuscated black box.
if (bed && bed_overlap(bed, h->target_name[last_tid], last_pos, last_pos + 1) == 0)
continue;
fputs(h->target_name[last_tid], stdout);
printf("\t%d", last_pos+1);
for (i = 0; i < n; i++)
putchar('\t'), putchar('0');
putchar('\n');
}
}
last_tid++;
last_pos = -1;
if (all < 2)
break;
}
// Deal with missing portion of current tid
while (++last_pos < pos) {
if (last_pos < beg) continue; // out of range; skip
if (bed && bed_overlap(bed, h->target_name[tid], last_pos, last_pos + 1) == 0)
continue;
fputs(h->target_name[tid], stdout);
printf("\t%d", last_pos+1);
for (i = 0; i < n; i++)
putchar('\t'), putchar('0');
putchar('\n');
}
last_tid = tid;
last_pos = pos;
}
if (bed && bed_overlap(bed, h->target_name[tid], pos, pos + 1) == 0) continue;
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 (bam_get_qual(p->b)[p->qpos] < baseQ) ++m; // low base quality
}
printf("\t%d", n_plp[i] - m); // this the depth to output
}
putchar('\n');
}
if (ret < 0) status = EXIT_FAILURE;
free(n_plp);
free(plp);
bam_mplp_destroy(mplp);
if (all) {
// Handle terminating region
if (last_tid < 0 && reg && all > 1) {
last_tid = reg_tid;
last_pos = beg-1;
}
while (last_tid >= 0 && last_tid < h->n_targets) {
while (++last_pos < h->target_len[last_tid]) {
if (last_pos >= end) break;
if (bed && bed_overlap(bed, h->target_name[last_tid], last_pos, last_pos + 1) == 0)
continue;
fputs(h->target_name[last_tid], stdout);
printf("\t%d", last_pos+1);
for (i = 0; i < n; i++)
putchar('\t'), putchar('0');
putchar('\n');
}
last_tid++;
last_pos = -1;
if (all < 2 || reg)
break;
}
}
depth_end:
for (i = 0; i < n && data[i]; ++i) {
bam_hdr_destroy(data[i]->hdr);
if (data[i]->fp) sam_close(data[i]->fp);
hts_itr_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);
}
sam_global_args_free(&ga);
return status;
}
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_mpileup(int argc, char *argv[])
{
int c;
const char *file_list = NULL;
char **fn = NULL;
int nfiles = 0, use_orphan = 0, noref = 0;
mplp_conf_t mplp;
memset(&mplp, 0, sizeof(mplp_conf_t));
mplp.min_baseQ = 13;
mplp.capQ_thres = 0;
mplp.max_depth = 250; mplp.max_indel_depth = 250;
mplp.openQ = 40; mplp.extQ = 20; mplp.tandemQ = 100;
mplp.min_frac = 0.002; mplp.min_support = 1;
mplp.flag = MPLP_NO_ORPHAN | MPLP_REALN | MPLP_SMART_OVERLAPS;
mplp.argc = argc; mplp.argv = argv;
mplp.rflag_filter = BAM_FUNMAP | BAM_FSECONDARY | BAM_FQCFAIL | BAM_FDUP;
mplp.output_fname = NULL;
mplp.output_type = FT_VCF;
mplp.record_cmd_line = 1;
mplp.n_threads = 0;
mplp.bsmpl = bam_smpl_init();
static const struct option lopts[] =
{
{"rf", required_argument, NULL, 1}, // require flag
{"ff", required_argument, NULL, 2}, // filter flag
{"incl-flags", required_argument, NULL, 1},
{"excl-flags", required_argument, NULL, 2},
{"output", required_argument, NULL, 3},
{"open-prob", required_argument, NULL, 4},
{"ignore-RG", no_argument, NULL, 5},
{"ignore-rg", no_argument, NULL, 5},
{"gvcf", required_argument, NULL, 'g'},
{"no-reference", no_argument, NULL, 7},
{"no-version", no_argument, NULL, 8},
{"threads",required_argument,NULL,9},
{"illumina1.3+", no_argument, NULL, '6'},
{"count-orphans", no_argument, NULL, 'A'},
{"bam-list", required_argument, NULL, 'b'},
{"no-BAQ", no_argument, NULL, 'B'},
{"no-baq", no_argument, NULL, 'B'},
{"adjust-MQ", required_argument, NULL, 'C'},
{"adjust-mq", required_argument, NULL, 'C'},
{"max-depth", required_argument, NULL, 'd'},
{"redo-BAQ", no_argument, NULL, 'E'},
{"redo-baq", no_argument, NULL, 'E'},
{"fasta-ref", required_argument, NULL, 'f'},
{"read-groups", required_argument, NULL, 'G'},
{"region", required_argument, NULL, 'r'},
{"regions", required_argument, NULL, 'r'},
{"regions-file", required_argument, NULL, 'R'},
{"targets", required_argument, NULL, 't'},
{"targets-file", required_argument, NULL, 'T'},
{"min-MQ", required_argument, NULL, 'q'},
{"min-mq", required_argument, NULL, 'q'},
{"min-BQ", required_argument, NULL, 'Q'},
{"min-bq", required_argument, NULL, 'Q'},
{"ignore-overlaps", no_argument, NULL, 'x'},
{"output-type", required_argument, NULL, 'O'},
{"samples", required_argument, NULL, 's'},
{"samples-file", required_argument, NULL, 'S'},
{"annotate", required_argument, NULL, 'a'},
{"ext-prob", required_argument, NULL, 'e'},
{"gap-frac", required_argument, NULL, 'F'},
{"tandem-qual", required_argument, NULL, 'h'},
{"skip-indels", no_argument, NULL, 'I'},
{"max-idepth", required_argument, NULL, 'L'},
{"min-ireads ", required_argument, NULL, 'm'},
{"per-sample-mF", no_argument, NULL, 'p'},
{"per-sample-mf", no_argument, NULL, 'p'},
{"platforms", required_argument, NULL, 'P'},
{NULL, 0, NULL, 0}
};
while ((c = getopt_long(argc, argv, "Ag:f:r:R:q:Q:C:Bd:L:b:P:po:e:h:Im:F:EG:6O:xa:s:S:t:T:",lopts,NULL)) >= 0) {
switch (c) {
case 'x': mplp.flag &= ~MPLP_SMART_OVERLAPS; break;
case 1 :
mplp.rflag_require = bam_str2flag(optarg);
if ( mplp.rflag_require<0 ) { fprintf(stderr,"Could not parse --rf %s\n", optarg); return 1; }
break;
case 2 :
mplp.rflag_filter = bam_str2flag(optarg);
if ( mplp.rflag_filter<0 ) { fprintf(stderr,"Could not parse --ff %s\n", optarg); return 1; }
break;
case 3 : mplp.output_fname = optarg; break;
case 4 : mplp.openQ = atoi(optarg); break;
case 5 : bam_smpl_ignore_readgroups(mplp.bsmpl); break;
case 'g':
mplp.gvcf = gvcf_init(optarg);
if ( !mplp.gvcf ) error("Could not parse: --gvcf %s\n", optarg);
break;
case 'f':
mplp.fai = fai_load(optarg);
if (mplp.fai == NULL) return 1;
mplp.fai_fname = optarg;
break;
case 7 : noref = 1; break;
case 8 : mplp.record_cmd_line = 0; break;
case 9 : mplp.n_threads = strtol(optarg, 0, 0); break;
case 'd': mplp.max_depth = atoi(optarg); break;
case 'r': mplp.reg_fname = strdup(optarg); break;
case 'R': mplp.reg_fname = strdup(optarg); mplp.reg_is_file = 1; break;
case 't':
// In the original version the whole BAM was streamed which is inefficient
// with few BED intervals and big BAMs. Todo: devise a heuristic to determine
// best strategy, that is streaming or jumping.
if ( optarg[0]=='^' ) optarg++;
else mplp.bed_logic = 1;
mplp.bed = regidx_init(NULL,regidx_parse_reg,NULL,0,NULL);
mplp.bed_itr = regitr_init(mplp.bed);
if ( regidx_insert_list(mplp.bed,optarg,',') !=0 )
{
fprintf(stderr,"Could not parse the targets: %s\n", optarg);
exit(EXIT_FAILURE);
}
break;
case 'T':
if ( optarg[0]=='^' ) optarg++;
else mplp.bed_logic = 1;
mplp.bed = regidx_init(optarg,NULL,NULL,0,NULL);
if (!mplp.bed) { fprintf(stderr, "bcftools mpileup: Could not read file \"%s\"", optarg); return 1; }
break;
case 'P': mplp.pl_list = strdup(optarg); break;
case 'p': mplp.flag |= MPLP_PER_SAMPLE; break;
case 'B': mplp.flag &= ~MPLP_REALN; break;
case 'I': mplp.flag |= MPLP_NO_INDEL; break;
case 'E': mplp.flag |= MPLP_REDO_BAQ; break;
case '6': mplp.flag |= MPLP_ILLUMINA13; break;
case 's': if ( bam_smpl_add_samples(mplp.bsmpl,optarg,0)<0 ) error("Could not read samples: %s\n",optarg); break;
case 'S': if ( bam_smpl_add_samples(mplp.bsmpl,optarg,1)<0 ) error("Could not read samples: %s\n",optarg); break;
case 'O':
switch (optarg[0]) {
case 'b': mplp.output_type = FT_BCF_GZ; break;
case 'u': mplp.output_type = FT_BCF; break;
case 'z': mplp.output_type = FT_VCF_GZ; break;
case 'v': mplp.output_type = FT_VCF; break;
default: error("[error] The option \"-O\" changed meaning when mpileup moved to bcftools. Did you mean: \"bcftools mpileup --output-type\" or \"samtools mpileup --output-BP\"?\n");
}
break;
case 'C': mplp.capQ_thres = atoi(optarg); break;
case 'q': mplp.min_mq = atoi(optarg); break;
case 'Q': mplp.min_baseQ = atoi(optarg); break;
case 'b': file_list = optarg; break;
case 'o': {
char *end;
long value = strtol(optarg, &end, 10);
// Distinguish between -o INT and -o FILE (a bit of a hack!)
if (*end == '\0') mplp.openQ = value;
else mplp.output_fname = optarg;
}
break;
case 'e': mplp.extQ = atoi(optarg); break;
case 'h': mplp.tandemQ = atoi(optarg); break;
case 'A': use_orphan = 1; break;
case 'F': mplp.min_frac = atof(optarg); break;
case 'm': mplp.min_support = atoi(optarg); break;
case 'L': mplp.max_indel_depth = atoi(optarg); break;
case 'G': bam_smpl_add_readgroups(mplp.bsmpl, optarg, 1); break;
case 'a':
if (optarg[0]=='?') {
list_annotations(stderr);
return 1;
}
mplp.fmt_flag |= parse_format_flag(optarg);
break;
default:
fprintf(stderr,"Invalid option: '%c'\n", c);
return 1;
}
}
if ( mplp.gvcf && !(mplp.fmt_flag&B2B_FMT_DP) )
{
fprintf(stderr,"[warning] The -t DP option is required with --gvcf, switching on.\n");
mplp.fmt_flag |= B2B_FMT_DP;
}
if ( mplp.flag&(MPLP_BCF|MPLP_VCF|MPLP_NO_COMP) )
{
if ( mplp.flag&MPLP_VCF )
{
if ( mplp.flag&MPLP_NO_COMP ) mplp.output_type = FT_VCF;
else mplp.output_type = FT_VCF_GZ;
}
else if ( mplp.flag&MPLP_BCF )
{
if ( mplp.flag&MPLP_NO_COMP ) mplp.output_type = FT_BCF;
else mplp.output_type = FT_BCF_GZ;
}
}
if ( !(mplp.flag&MPLP_REALN) && mplp.flag&MPLP_REDO_BAQ )
{
fprintf(stderr,"Error: The -B option cannot be combined with -E\n");
return 1;
}
if (use_orphan) mplp.flag &= ~MPLP_NO_ORPHAN;
if (argc == 1)
{
print_usage(stderr, &mplp);
return 1;
}
if (!mplp.fai && !noref) {
fprintf(stderr,"Error: mpileup requires the --fasta-ref option by default; use --no-reference to run without a fasta reference\n");
return 1;
}
int ret,i;
if (file_list)
{
if ( read_file_list(file_list,&nfiles,&fn) ) return 1;
mplp.files = fn;
mplp.nfiles = nfiles;
}
else
{
mplp.nfiles = argc - optind;
mplp.files = (char**) malloc(mplp.nfiles*sizeof(char*));
for (i=0; i<mplp.nfiles; i++) mplp.files[i] = strdup(argv[optind+i]);
}
ret = mpileup(&mplp);
for (i=0; i<mplp.nfiles; i++) free(mplp.files[i]);
free(mplp.files);
free(mplp.reg_fname); free(mplp.pl_list);
if (mplp.fai) fai_destroy(mplp.fai);
if (mplp.bed) regidx_destroy(mplp.bed);
if (mplp.bed_itr) regitr_destroy(mplp.bed_itr);
if (mplp.reg) regidx_destroy(mplp.reg);
bam_smpl_destroy(mplp.bsmpl);
return ret;
}
int bam_mpileup(int argc, char *argv[])
{
int c;
const char *file_list = NULL;
char **fn = NULL;
int nfiles = 0, use_orphan = 0;
mplp_conf_t mplp;
memset(&mplp, 0, sizeof(mplp_conf_t));
mplp.min_baseQ = 13;
mplp.capQ_thres = 0;
mplp.max_depth = 250; mplp.max_indel_depth = 250;
mplp.openQ = 40; mplp.extQ = 20; mplp.tandemQ = 100;
mplp.min_frac = 0.002; mplp.min_support = 1;
mplp.flag = MPLP_NO_ORPHAN | MPLP_REALN | MPLP_SMART_OVERLAPS;
mplp.argc = argc; mplp.argv = argv;
mplp.rflag_filter = BAM_FUNMAP | BAM_FSECONDARY | BAM_FQCFAIL | BAM_FDUP;
mplp.output_fname = NULL;
static const struct option lopts[] =
{
{"rf", required_argument, NULL, 1}, // require flag
{"ff", required_argument, NULL, 2}, // filter flag
{"incl-flags", required_argument, NULL, 1},
{"excl-flags", required_argument, NULL, 2},
{"output", required_argument, NULL, 3},
{"open-prob", required_argument, NULL, 4},
{"illumina1.3+", no_argument, NULL, '6'},
{"count-orphans", no_argument, NULL, 'A'},
{"bam-list", required_argument, NULL, 'b'},
{"no-BAQ", no_argument, NULL, 'B'},
{"no-baq", no_argument, NULL, 'B'},
{"adjust-MQ", required_argument, NULL, 'C'},
{"adjust-mq", required_argument, NULL, 'C'},
{"max-depth", required_argument, NULL, 'd'},
{"redo-BAQ", no_argument, NULL, 'E'},
{"redo-baq", no_argument, NULL, 'E'},
{"fasta-ref", required_argument, NULL, 'f'},
{"exclude-RG", required_argument, NULL, 'G'},
{"exclude-rg", required_argument, NULL, 'G'},
{"positions", required_argument, NULL, 'l'},
{"region", required_argument, NULL, 'r'},
{"ignore-RG", no_argument, NULL, 'R'},
{"ignore-rg", no_argument, NULL, 'R'},
{"min-MQ", required_argument, NULL, 'q'},
{"min-mq", required_argument, NULL, 'q'},
{"min-BQ", required_argument, NULL, 'Q'},
{"min-bq", required_argument, NULL, 'Q'},
{"ignore-overlaps", no_argument, NULL, 'x'},
{"BCF", no_argument, NULL, 'g'},
{"bcf", no_argument, NULL, 'g'},
{"VCF", no_argument, NULL, 'v'},
{"vcf", no_argument, NULL, 'v'},
{"output-BP", no_argument, NULL, 'O'},
{"output-bp", no_argument, NULL, 'O'},
{"output-MQ", no_argument, NULL, 's'},
{"output-mq", no_argument, NULL, 's'},
{"output-tags", required_argument, NULL, 't'},
{"uncompressed", no_argument, NULL, 'u'},
{"ext-prob", required_argument, NULL, 'e'},
{"gap-frac", required_argument, NULL, 'F'},
{"tandem-qual", required_argument, NULL, 'h'},
{"skip-indels", no_argument, NULL, 'I'},
{"max-idepth", required_argument, NULL, 'L'},
{"min-ireads ", required_argument, NULL, 'm'},
{"per-sample-mF", no_argument, NULL, 'p'},
{"per-sample-mf", no_argument, NULL, 'p'},
{"platforms", required_argument, NULL, 'P'},
{NULL, 0, NULL, 0}
};
while ((c = getopt_long(argc, argv, "Agf:r:l:q:Q:uRC:BDSd:L:b:P:po:e:h:Im:F:EG:6OsVvxt:",lopts,NULL)) >= 0) {
switch (c) {
case 'x': mplp.flag &= ~MPLP_SMART_OVERLAPS; break;
case 1 :
mplp.rflag_require = bam_str2flag(optarg);
if ( mplp.rflag_require<0 ) { fprintf(stderr,"Could not parse --rf %s\n", optarg); return 1; }
break;
case 2 :
mplp.rflag_filter = bam_str2flag(optarg);
if ( mplp.rflag_filter<0 ) { fprintf(stderr,"Could not parse --ff %s\n", optarg); return 1; }
break;
case 3 : mplp.output_fname = optarg; break;
case 4 : mplp.openQ = atoi(optarg); break;
case 'f':
mplp.fai = fai_load(optarg);
if (mplp.fai == 0) return 1;
mplp.fai_fname = optarg;
break;
case 'd': mplp.max_depth = atoi(optarg); break;
case 'r': mplp.reg = strdup(optarg); break;
case 'l':
// In the original version the whole BAM was streamed which is inefficient
// with few BED intervals and big BAMs. Todo: devise a heuristic to determine
// best strategy, that is streaming or jumping.
mplp.bed = bed_read(optarg);
if (!mplp.bed) { print_error_errno("Could not read file \"%s\"", optarg); return 1; }
break;
case 'P': mplp.pl_list = strdup(optarg); break;
case 'p': mplp.flag |= MPLP_PER_SAMPLE; break;
case 'g': mplp.flag |= MPLP_BCF; break;
case 'v': mplp.flag |= MPLP_BCF | MPLP_VCF; break;
case 'u': mplp.flag |= MPLP_NO_COMP | MPLP_BCF; break;
case 'B': mplp.flag &= ~MPLP_REALN; break;
case 'D': mplp.fmt_flag |= B2B_FMT_DP; fprintf(stderr, "[warning] samtools mpileup option `-D` is functional, but deprecated. Please switch to `-t DP` in future.\n"); break;
case 'S': mplp.fmt_flag |= B2B_FMT_SP; fprintf(stderr, "[warning] samtools mpileup option `-S` is functional, but deprecated. Please switch to `-t SP` in future.\n"); break;
case 'V': mplp.fmt_flag |= B2B_FMT_DV; fprintf(stderr, "[warning] samtools mpileup option `-V` is functional, but deprecated. Please switch to `-t DV` in future.\n"); break;
case 'I': mplp.flag |= MPLP_NO_INDEL; break;
case 'E': mplp.flag |= MPLP_REDO_BAQ; break;
case '6': mplp.flag |= MPLP_ILLUMINA13; break;
case 'R': mplp.flag |= MPLP_IGNORE_RG; break;
case 's': mplp.flag |= MPLP_PRINT_MAPQ; break;
case 'O': mplp.flag |= MPLP_PRINT_POS; break;
case 'C': mplp.capQ_thres = atoi(optarg); break;
case 'q': mplp.min_mq = atoi(optarg); break;
case 'Q': mplp.min_baseQ = atoi(optarg); break;
case 'b': file_list = optarg; break;
case 'o': {
char *end;
long value = strtol(optarg, &end, 10);
// Distinguish between -o INT and -o FILE (a bit of a hack!)
if (*end == '\0') mplp.openQ = value;
else mplp.output_fname = optarg;
}
break;
case 'e': mplp.extQ = atoi(optarg); break;
case 'h': mplp.tandemQ = atoi(optarg); break;
case 'A': use_orphan = 1; break;
case 'F': mplp.min_frac = atof(optarg); break;
case 'm': mplp.min_support = atoi(optarg); break;
case 'L': mplp.max_indel_depth = atoi(optarg); break;
case 'G': {
FILE *fp_rg;
char buf[1024];
mplp.rghash = khash_str2int_init();
if ((fp_rg = fopen(optarg, "r")) == 0)
fprintf(stderr, "(%s) Fail to open file %s. Continue anyway.\n", __func__, optarg);
while (!feof(fp_rg) && fscanf(fp_rg, "%s", buf) > 0) // this is not a good style, but forgive me...
khash_str2int_inc(mplp.rghash, strdup(buf));
fclose(fp_rg);
}
break;
case 't': mplp.fmt_flag |= parse_format_flag(optarg); break;
default:
fprintf(stderr,"Invalid option: '%c'\n", c);
return 1;
}
}
if ( !(mplp.flag&MPLP_REALN) && mplp.flag&MPLP_REDO_BAQ )
{
fprintf(stderr,"Error: The -B option cannot be combined with -E\n");
return 1;
}
if (use_orphan) mplp.flag &= ~MPLP_NO_ORPHAN;
if (argc == 1)
{
print_usage(stderr, &mplp);
return 1;
}
int ret;
if (file_list) {
if ( read_file_list(file_list,&nfiles,&fn) ) return 1;
ret = mpileup(&mplp,nfiles,fn);
for (c=0; c<nfiles; c++) free(fn[c]);
free(fn);
}
else
ret = mpileup(&mplp, argc - optind, argv + optind);
if (mplp.rghash) khash_str2int_destroy_free(mplp.rghash);
free(mplp.reg); free(mplp.pl_list);
if (mplp.fai) fai_destroy(mplp.fai);
if (mplp.bed) bed_destroy(mplp.bed);
return ret;
}
int main_depth(int argc, char *argv[])
{
int i, n, tid, beg, end, pos, *n_plp, baseQ = 0, mapQ = 0, min_len = 0, status = EXIT_SUCCESS, nfiles;
const bam_pileup1_t **plp;
char *reg = 0; // specified region
void *bed = 0; // BED data structure
char *file_list = NULL, **fn = NULL;
bam_hdr_t *h = NULL; // 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); // BED or position list file can be parsed now
if (!bed) { print_error_errno("Could not read file \"%s\"", optarg); return 1; }
break;
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(pysamerr, "\n");
fprintf(pysamerr, "Usage: samtools depth [options] in1.bam [in2.bam [...]]\n");
fprintf(pysamerr, "Options:\n");
fprintf(pysamerr, " -b <bed> list of positions or regions\n");
fprintf(pysamerr, " -f <list> list of input BAM filenames, one per line [null]\n");
fprintf(pysamerr, " -l <int> read length threshold (ignore reads shorter than <int>)\n");
fprintf(pysamerr, " -q <int> base quality threshold\n");
fprintf(pysamerr, " -Q <int> mapping quality threshold\n");
fprintf(pysamerr, " -r <chr:from-to> region\n");
fprintf(pysamerr, "\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(aux_t*)); // data[i] for the i-th input
beg = 0; end = 1<<30; // set the default region
for (i = 0; i < n; ++i) {
data[i] = calloc(1, sizeof(aux_t));
data[i]->fp = sam_open(argv[optind+i], "r"); // open BAM
if (data[i]->fp == NULL) {
print_error_errno("Could not open \"%s\"", argv[optind+i]);
status = EXIT_FAILURE;
goto depth_end;
}
if (hts_set_opt(data[i]->fp, CRAM_OPT_REQUIRED_FIELDS,
SAM_FLAG | SAM_RNAME | SAM_POS | SAM_MAPQ | SAM_CIGAR |
SAM_SEQ)) {
fprintf(pysamerr, "Failed to set CRAM_OPT_REQUIRED_FIELDS value\n");
return 1;
}
if (hts_set_opt(data[i]->fp, CRAM_OPT_DECODE_MD, 0)) {
fprintf(pysamerr, "Failed to set CRAM_OPT_DECODE_MD value\n");
return 1;
}
data[i]->min_mapQ = mapQ; // set the mapQ filter
data[i]->min_len = min_len; // set the qlen filter
data[i]->hdr = sam_hdr_read(data[i]->fp); // read the BAM header
if (reg) { // if a region is specified
hts_idx_t *idx = sam_index_load(data[i]->fp, argv[optind+i]); // load the index
if (idx == NULL) {
print_error("can't load index for \"%s\"", argv[optind+i]);
status = EXIT_FAILURE;
goto depth_end;
}
data[i]->iter = sam_itr_querys(idx, data[i]->hdr, reg); // set the iterator
hts_idx_destroy(idx); // the index is not needed any more; free the memory
if (data[i]->iter == NULL) {
print_error("can't parse region \"%s\"", reg);
status = EXIT_FAILURE;
goto depth_end;
}
}
}
h = data[0]->hdr; // easy access to the header of the 1st BAM
if (reg) {
beg = data[0]->iter->beg; // and to the parsed region coordinates
end = data[0]->iter->end;
}
// the core multi-pileup loop
mplp = bam_mplp_init(n, read_bam, (void**)data); // initialization
n_plp = calloc(n, sizeof(int)); // n_plp[i] is the number of covering reads from the i-th BAM
plp = calloc(n, sizeof(bam_pileup1_t*)); // 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 (bam_get_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);
depth_end:
for (i = 0; i < n && data[i]; ++i) {
bam_hdr_destroy(data[i]->hdr);
if (data[i]->fp) sam_close(data[i]->fp);
hts_itr_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 status;
}
static int read_directory(const char * dname)
{
int ret = 0;
solist_t files;
solist_t dirs;
strobj_t sobj;
//d_dbg("%s: dname = '%s'\n",__func__,dname);
do
{
files = solist_new();
dirs = solist_new();
if (!files || !dirs)
{
ret = -1;
verbose("solist_new() return error!\n");
break;
}
ret = read_file_list(dname, files, dirs);
if (ret < 0)
{
verbose("error reading file list @ '%s' !\n", dname);
break;
}
/* handle files first */
ret = 0;
while ((sobj=solist_get_next(files)) != NULL)
{
ret = read_file(sobj_get_string(sobj));
if (ret < 0)
{
verbose("error handling file - '%s'\n",sobj_get_string(sobj));
break;
}
}
if (ret < 0) break;
/* handle diretories if recursive. */
if (o_recursive)
{
ret = 0;
while ((sobj=solist_get_next(dirs)) != NULL)
{
ret = read_directory(sobj_get_string(sobj));
if (ret < 0)
{
verbose("error handling directory - '%s'\n",sobj_get_string(sobj));
break;
}
}
if (ret < 0) break;
}
} while (0);
if (files) solist_del(files);
if (dirs) solist_del(dirs);
return ret;
}