static void copy_check_alignment(const char *infname, const char *informat,
const char *outfname, const char *outmode, const char *outref)
{
samFile *in = sam_open(infname, "r");
samFile *out = sam_open(outfname, outmode);
bam1_t *aln = bam_init1();
bam_hdr_t *header = NULL;
int res;
if (!in) {
fail("couldn't open %s", infname);
goto err;
}
if (!out) {
fail("couldn't open %s with mode %s", outfname, outmode);
goto err;
}
if (!aln) {
fail("bam_init1() failed");
goto err;
}
if (outref) {
if (hts_set_opt(out, CRAM_OPT_REFERENCE, outref) < 0) {
fail("setting reference %s for %s", outref, outfname);
goto err;
}
}
header = sam_hdr_read(in);
if (!header) {
fail("reading header from %s", infname);
goto err;
}
if (sam_hdr_write(out, header) < 0) fail("writing headers to %s", outfname);
while ((res = sam_read1(in, header, aln)) >= 0) {
int mod4 = ((intptr_t) bam_get_cigar(aln)) % 4;
if (mod4 != 0)
fail("%s CIGAR not 4-byte aligned; offset is 4k+%d for \"%s\"",
informat, mod4, bam_get_qname(aln));
if (sam_write1(out, header, aln) < 0) fail("writing to %s", outfname);
}
if (res < -1) {
fail("failed to read alignment from %s", infname);
}
err:
bam_destroy1(aln);
bam_hdr_destroy(header);
if (in) sam_close(in);
if (out) sam_close(out);
}
static bool readgroupise(state_t* state)
{
if (sam_hdr_write(state->output_file, state->output_header) != 0) {
print_error_errno("addreplacerg", "[%s] Could not write header to output file", __func__);
return false;
}
bam1_t* file_read = bam_init1();
int ret;
while ((ret = sam_read1(state->input_file, state->input_header, file_read)) >= 0) {
state->mode_func(state, file_read);
if (sam_write1(state->output_file, state->output_header, file_read) < 0) {
print_error_errno("addreplacerg", "[%s] Could not write read to output file", __func__);
bam_destroy1(file_read);
return false;
}
}
bam_destroy1(file_read);
if (ret != -1) {
print_error_errno("addreplacerg", "[%s] Error reading from input file", __func__);
return false;
} else {
return true;
}
}
int main(int argc, char **argv) {
dlib::BamHandle in = dlib::BamHandle("bed_test.bam");
dlib::ParsedBed bed = dlib::ParsedBed("bed_test.bed", in.header);
bam1_t *b = bam_init1();
size_t diffs = 0;
void *lh3bed = bed_read("bed_test.bed");
samFile *so = sam_open("disagreed.bam", "wb9");
sam_hdr_write(so, in.header);
size_t disagrees = 0, agrees = 0;
int dbr = 0, lh3r = 0;
while(in.read(b) != -1) {
if(b->core.flag & (BAM_FUNMAP)) continue;
if((dbr = bed.bam1_test(b)) != (lh3r = bed_overlap(lh3bed, in.header->target_name[b->core.tid], b->core.pos, bam_endpos(b)))) {
LOG_EXIT("dbr: %i. lh3r: %i. Contig: %s. Position: %i. endpos; %i\n", dbr, lh3r, in.header->target_name[b->core.tid], b->core.pos, bam_endpos(b));
if(++disagrees % 100 == 0) LOG_DEBUG("disagrees: %lu.\n", disagrees);
sam_write1(so, in.header, b);
} else {
if(++agrees % 500000 == 0) LOG_DEBUG("agrees: %lu.\n", agrees);
}
}
sam_close(so);
bam_destroy1(b);
bed_destroy(lh3bed);
return EXIT_SUCCESS;
}
void ingestHeader(std::shared_ptr<bam_hdr_t> &header) {
auto version = bamql::version();
std::stringstream name;
name << "bamql-chain ";
for (auto chains = known_chains.begin(); chains != known_chains.end();
chains++) {
if (chains->second == chain) {
name << chains->first;
break;
}
}
uuid_t uuid;
uuid_generate(uuid);
char id_str[sizeof(uuid_t) * 2 + 1];
uuid_unparse(uuid, id_str);
auto copy = bamql::appendProgramToHeader(
header.get(), name.str(), std::string(id_str), version, query);
if (output_file) {
sam_hdr_write(output_file.get(), copy.get());
}
if (next)
next->ingestHeader(chain == 3 ? header : copy);
}
/*
* Reads a file and outputs a new CRAM file to stdout with 'h'
* replaced as the header. No checks are made to the validity.
*
* FIXME: error checking
*/
int cram_reheader(cram_fd *in, bam_hdr_t *h, const char *arg_list, int add_PG)
{
htsFile *h_out = hts_open("-", "wc");
cram_fd *out = h_out->fp.cram;
cram_container *c = NULL;
int ret = -1;
// Attempt to fill out a cram->refs[] array from @SQ headers
cram_fd_set_header(out, sam_hdr_parse_(h->text, h->l_text));
if (add_PG) {
if (sam_hdr_add_PG(cram_fd_get_header(out), "samtools",
"VN", samtools_version(),
arg_list ? "CL": NULL,
arg_list ? arg_list : NULL,
NULL) != 0)
goto err;
// Covert back to bam_hdr_t struct
free(h->text);
h->text = strdup(sam_hdr_str(cram_fd_get_header(out)));
h->l_text = sam_hdr_length(cram_fd_get_header(out));
if (!h->text)
goto err;
}
if (sam_hdr_write(h_out, h) != 0)
goto err;
cram_set_option(out, CRAM_OPT_REFERENCE, NULL);
while ((c = cram_read_container(in))) {
int32_t i, num_blocks = cram_container_get_num_blocks(c);
if (cram_write_container(out, c) != 0)
goto err;
for (i = 0; i < num_blocks; i++) {
cram_block *blk = cram_read_block(in);
if (!blk || cram_write_block(out, blk) != 0) {
if (blk) cram_free_block(blk);
goto err;
}
cram_free_block(blk);
}
cram_free_container(c);
}
ret = 0;
err:
if (hts_close(h_out) != 0)
ret = -1;
return ret;
}
static void write_buffer(const char *fn, const char *mode, size_t l, bam1_p *buf, const bam_hdr_t *h, int n_threads)
{
size_t i;
samFile* fp;
fp = sam_open(fn, mode);
if (fp == NULL) return;
sam_hdr_write(fp, h);
if (n_threads > 1) hts_set_threads(fp, n_threads);
for (i = 0; i < l; ++i)
sam_write1(fp, h, buf[i]);
sam_close(fp);
}
samfile_t *samopen(const char *fn, const char *mode, const void *aux)
{
// hts_open() is really sam_open(), except for #define games
samFile *hts_fp = hts_open(fn, mode);
if (hts_fp == NULL) return NULL;
samfile_t *fp = malloc(sizeof (samfile_t));
if (!fp) {
sam_close(hts_fp);
return NULL;
}
fp->file = hts_fp;
fp->x.bam = hts_fp->fp.bgzf;
if (strchr(mode, 'r')) {
if (aux) {
if (hts_set_fai_filename(fp->file, aux) != 0) {
sam_close(hts_fp);
free(fp);
return NULL;
}
}
fp->header = sam_hdr_read(fp->file); // samclose() will free this
if (fp->header == NULL) {
sam_close(hts_fp);
free(fp);
return NULL;
}
fp->is_write = 0;
if (fp->header->n_targets == 0 && bam_verbose >= 1)
fprintf(samtools_stderr, "[samopen] no @SQ lines in the header.\n");
}
else {
enum htsExactFormat fmt = hts_get_format(fp->file)->format;
fp->header = (bam_hdr_t *)aux; // For writing, we won't free it
fp->is_write = 1;
if (!(fmt == text_format || fmt == sam) || strchr(mode, 'h')) {
if (sam_hdr_write(fp->file, fp->header) < 0) {
if (bam_verbose >= 1)
fprintf(samtools_stderr, "[samopen] Couldn't write header\n");
sam_close(hts_fp);
free(fp);
return NULL;
}
}
}
return fp;
}
static int view_sam(hFILE *hfp, const char *filename)
{
samFile *in = hts_hopen(hfp, filename, "r");
if (in == NULL) return 0;
samFile *out = dup_stdout("w");
bam_hdr_t *hdr = sam_hdr_read(in);
if (show_headers) sam_hdr_write(out, hdr);
if (mode == view_all) {
bam1_t *b = bam_init1();
while (sam_read1(in, hdr, b) >= 0)
sam_write1(out, hdr, b);
bam_destroy1(b);
}
bam_hdr_destroy(hdr);
hts_close(out);
hts_close(in);
return 1;
}
void parse() {
if (sam_hdr_write(out_file, replace_header) != 0) {
throw new std::runtime_error("IEEEE!");
}
bam1_t* file_read = bam_init1();
while (sam_read1(file_iter, file_header, file_read) >= 0) {
if (file_read->core.tid != -1) {
file_read->core.tid = trans[file_read->core.tid];
}
if (file_read->core.mtid != -1) {
file_read->core.mtid = trans[file_read->core.mtid];
}
sam_write1(out_file, file_header, file_read);
}
// Clean up
if (file_read) { bam_destroy1(file_read); }
}
SamWriterPrivate(const std::string& filename,
const PBBAM_SHARED_PTR<bam_hdr_t> rawHeader)
: internal::FileProducer(filename)
, file_(nullptr)
, header_(rawHeader)
{
if (!header_)
throw std::runtime_error("null header");
// open file
const string& usingFilename = TempFilename();
const string& mode = string("w");
file_.reset(sam_open(usingFilename.c_str(), mode.c_str()));
if (!file_)
throw std::runtime_error("could not open file for writing");
// write header
const int ret = sam_hdr_write(file_.get(), header_.get());
if (ret != 0)
throw std::runtime_error("could not write header");
}
void SamWriter::write_header() const {
sam_hdr_write(m_out_file, m_header.m_header.get());
}
int main_samview(int argc, char *argv[])
{
int index;
for(index = 0; index < argc; index++) {
printf("The %d is %s\n",index,argv[index]);
}
getchar();return 0;
int c, is_header = 0, is_header_only = 0, ret = 0, compress_level = -1, is_count = 0;
int is_long_help = 0, n_threads = 0;
int64_t count = 0;
samFile *in = 0, *out = 0, *un_out=0;
bam_hdr_t *header = NULL;
char out_mode[5], out_un_mode[5], *out_format = "";
char *fn_in = 0, *fn_out = 0, *fn_list = 0, *q, *fn_un_out = 0;
sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
samview_settings_t settings = {
.rghash = NULL,
.min_mapQ = 0,
.flag_on = 0,
.flag_off = 0,
.min_qlen = 0,
.remove_B = 0,
.subsam_seed = 0,
.subsam_frac = -1.,
.library = NULL,
.bed = NULL,
};
static const struct option lopts[] = {
SAM_OPT_GLOBAL_OPTIONS('-', 0, 'O', 0, 'T'),
{ "threads", required_argument, NULL, '@' },
{ NULL, 0, NULL, 0 }
};
/* parse command-line options */
strcpy(out_mode, "w");
strcpy(out_un_mode, "w");
while ((c = getopt_long(argc, argv,
"SbBcCt:h1Ho:O:q:f:F:ul:r:?T:R:L:s:@:m:x:U:",
lopts, NULL)) >= 0) {
switch (c) {
case 's':
if ((settings.subsam_seed = strtol(optarg, &q, 10)) != 0) {
srand(settings.subsam_seed);
settings.subsam_seed = rand();
}
settings.subsam_frac = strtod(q, &q);
break;
case 'm': settings.min_qlen = atoi(optarg); break;
case 'c': is_count = 1; break;
case 'S': break;
case 'b': out_format = "b"; break;
case 'C': out_format = "c"; break;
case 't': fn_list = strdup(optarg); break;
case 'h': is_header = 1; break;
case 'H': is_header_only = 1; break;
case 'o': fn_out = strdup(optarg); break;
case 'U': fn_un_out = strdup(optarg); break;
case 'f': settings.flag_on |= strtol(optarg, 0, 0); break;
case 'F': settings.flag_off |= strtol(optarg, 0, 0); break;
case 'q': settings.min_mapQ = atoi(optarg); break;
case 'u': compress_level = 0; break;
case '1': compress_level = 1; break;
case 'l': settings.library = strdup(optarg); break;
case 'L':
if ((settings.bed = bed_read(optarg)) == NULL) {
print_error_errno("view", "Could not read file \"%s\"", optarg);
ret = 1;
goto view_end;
}
break;
case 'r':
if (add_read_group_single("view", &settings, optarg) != 0) {
ret = 1;
goto view_end;
}
break;
case 'R':
if (add_read_groups_file("view", &settings, optarg) != 0) {
ret = 1;
goto view_end;
}
break;
/* REMOVED as htslib doesn't support this
//case 'x': out_format = "x"; break;
//case 'X': out_format = "X"; break;
*/
case '?': is_long_help = 1; break;
case 'B': settings.remove_B = 1; break;
case '@': n_threads = strtol(optarg, 0, 0); break;
case 'x':
{
if (strlen(optarg) != 2) {
fprintf(stderr, "main_samview: Error parsing -x auxiliary tags should be exactly two characters long.\n");
return usage(stderr, EXIT_FAILURE, is_long_help);
}
settings.remove_aux = (char**)realloc(settings.remove_aux, sizeof(char*) * (++settings.remove_aux_len));
settings.remove_aux[settings.remove_aux_len-1] = optarg;
}
break;
default:
if (parse_sam_global_opt(c, optarg, lopts, &ga) != 0)
return usage(stderr, EXIT_FAILURE, is_long_help);
break;
}
}
if (compress_level >= 0 && !*out_format) out_format = "b";
if (is_header_only) is_header = 1;
// File format auto-detection first
if (fn_out) sam_open_mode(out_mode+1, fn_out, NULL);
if (fn_un_out) sam_open_mode(out_un_mode+1, fn_un_out, NULL);
// Overridden by manual -b, -C
if (*out_format)
out_mode[1] = out_un_mode[1] = *out_format;
out_mode[2] = out_un_mode[2] = '\0';
// out_(un_)mode now 1 or 2 bytes long, followed by nul.
if (compress_level >= 0) {
char tmp[2];
tmp[0] = compress_level + '0'; tmp[1] = '\0';
strcat(out_mode, tmp);
strcat(out_un_mode, tmp);
}
if (argc == optind && isatty(STDIN_FILENO)) return usage(stdout, EXIT_SUCCESS, is_long_help); // potential memory leak...
fn_in = (optind < argc)? argv[optind] : "-";
// generate the fn_list if necessary
if (fn_list == 0 && ga.reference) fn_list = samfaipath(ga.reference);
// open file handlers
if ((in = sam_open_format(fn_in, "r", &ga.in)) == 0) {
print_error_errno("view", "failed to open \"%s\" for reading", fn_in);
ret = 1;
goto view_end;
}
if (fn_list) {
if (hts_set_fai_filename(in, fn_list) != 0) {
fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_list);
ret = 1;
goto view_end;
}
}
if ((header = sam_hdr_read(in)) == 0) {
fprintf(stderr, "[main_samview] fail to read the header from \"%s\".\n", fn_in);
ret = 1;
goto view_end;
}
if (settings.rghash) { // FIXME: I do not know what "bam_header_t::n_text" is for...
char *tmp;
int l;
tmp = drop_rg(header->text, settings.rghash, &l);
free(header->text);
header->text = tmp;
header->l_text = l;
}
if (!is_count) {
if ((out = sam_open_format(fn_out? fn_out : "-", out_mode, &ga.out)) == 0) {
print_error_errno("view", "failed to open \"%s\" for writing", fn_out? fn_out : "standard output");
ret = 1;
goto view_end;
}
if (fn_list) {
if (hts_set_fai_filename(out, fn_list) != 0) {
fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_list);
ret = 1;
goto view_end;
}
}
if (*out_format || is_header ||
out_mode[1] == 'b' || out_mode[1] == 'c' ||
(ga.out.format != sam && ga.out.format != unknown_format)) {
if (sam_hdr_write(out, header) != 0) {
fprintf(stderr, "[main_samview] failed to write the SAM header\n");
ret = 1;
goto view_end;
}
}
if (fn_un_out) {
if ((un_out = sam_open_format(fn_un_out, out_un_mode, &ga.out)) == 0) {
print_error_errno("view", "failed to open \"%s\" for writing", fn_un_out);
ret = 1;
goto view_end;
}
if (fn_list) {
if (hts_set_fai_filename(un_out, fn_list) != 0) {
fprintf(stderr, "[main_samview] failed to use reference \"%s\".\n", fn_list);
ret = 1;
goto view_end;
}
}
if (*out_format || is_header ||
out_un_mode[1] == 'b' || out_un_mode[1] == 'c' ||
(ga.out.format != sam && ga.out.format != unknown_format)) {
if (sam_hdr_write(un_out, header) != 0) {
fprintf(stderr, "[main_samview] failed to write the SAM header\n");
ret = 1;
goto view_end;
}
}
}
}
if (n_threads > 1) { if (out) hts_set_threads(out, n_threads); }
if (is_header_only) goto view_end; // no need to print alignments
if (optind + 1 >= argc) { // convert/print the entire file
bam1_t *b = bam_init1();
int r;
while ((r = sam_read1(in, header, b)) >= 0) { // read one alignment from `in'
if (!process_aln(header, b, &settings)) {
if (!is_count) { if (check_sam_write1(out, header, b, fn_out, &ret) < 0) break; }
count++;
} else {
if (un_out) { if (check_sam_write1(un_out, header, b, fn_un_out, &ret) < 0) break; }
}
}
if (r < -1) {
fprintf(stderr, "[main_samview] truncated file.\n");
ret = 1;
}
bam_destroy1(b);
} else { // retrieve alignments in specified regions
int i;
bam1_t *b;
hts_idx_t *idx = sam_index_load(in, fn_in); // load index
if (idx == 0) { // index is unavailable
fprintf(stderr, "[main_samview] random alignment retrieval only works for indexed BAM or CRAM files.\n");
ret = 1;
goto view_end;
}
b = bam_init1();
for (i = optind + 1; i < argc; ++i) {
int result;
hts_itr_t *iter = sam_itr_querys(idx, header, argv[i]); // parse a region in the format like `chr2:100-200'
if (iter == NULL) { // region invalid or reference name not found
int beg, end;
if (hts_parse_reg(argv[i], &beg, &end))
fprintf(stderr, "[main_samview] region \"%s\" specifies an unknown reference name. Continue anyway.\n", argv[i]);
else
fprintf(stderr, "[main_samview] region \"%s\" could not be parsed. Continue anyway.\n", argv[i]);
continue;
}
// fetch alignments
while ((result = sam_itr_next(in, iter, b)) >= 0) {
if (!process_aln(header, b, &settings)) {
if (!is_count) { if (check_sam_write1(out, header, b, fn_out, &ret) < 0) break; }
count++;
} else {
if (un_out) { if (check_sam_write1(un_out, header, b, fn_un_out, &ret) < 0) break; }
}
}
hts_itr_destroy(iter);
if (result < -1) {
fprintf(stderr, "[main_samview] retrieval of region \"%s\" failed due to truncated file or corrupt BAM index file\n", argv[i]);
ret = 1;
break;
}
}
bam_destroy1(b);
hts_idx_destroy(idx); // destroy the BAM index
}
view_end:
if (is_count && ret == 0)
printf("%" PRId64 "\n", count);
// close files, free and return
if (in) check_sam_close("view", in, fn_in, "standard input", &ret);
if (out) check_sam_close("view", out, fn_out, "standard output", &ret);
if (un_out) check_sam_close("view", un_out, fn_un_out, "file", &ret);
free(fn_list); free(fn_out); free(settings.library); free(fn_un_out);
sam_global_args_free(&ga);
if ( header ) bam_hdr_destroy(header);
if (settings.bed) bed_destroy(settings.bed);
if (settings.rghash) {
khint_t k;
for (k = 0; k < kh_end(settings.rghash); ++k)
if (kh_exist(settings.rghash, k)) free((char*)kh_key(settings.rghash, k));
kh_destroy(rg, settings.rghash);
}
if (settings.remove_aux_len) {
free(settings.remove_aux);
}
return ret;
}
static int usage(FILE *fp, int exit_status, int is_long_help)
{
fprintf(fp,
"\n"
"Usage: samtools view [options] <in.bam>|<in.sam>|<in.cram> [region ...]\n"
"\n"
"Options:\n"
// output options
" -b output BAM\n"
" -C output CRAM (requires -T)\n"
" -1 use fast BAM compression (implies -b)\n"
" -u uncompressed BAM output (implies -b)\n"
" -h include header in SAM output\n"
" -H print SAM header only (no alignments)\n"
" -c print only the count of matching records\n"
" -o FILE output file name [stdout]\n"
" -U FILE output reads not selected by filters to FILE [null]\n"
// extra input
" -t FILE FILE listing reference names and lengths (see long help) [null]\n"
// read filters
" -L FILE only include reads overlapping this BED FILE [null]\n"
" -r STR only include reads in read group STR [null]\n"
" -R FILE only include reads with read group listed in FILE [null]\n"
" -q INT only include reads with mapping quality >= INT [0]\n"
" -l STR only include reads in library STR [null]\n"
" -m INT only include reads with number of CIGAR operations consuming\n"
" query sequence >= INT [0]\n"
" -f INT only include reads with all bits set in INT set in FLAG [0]\n"
" -F INT only include reads with none of the bits set in INT set in FLAG [0]\n"
// read processing
" -x STR read tag to strip (repeatable) [null]\n"
" -B collapse the backward CIGAR operation\n"
" -s FLOAT integer part sets seed of random number generator [0];\n"
" rest sets fraction of templates to subsample [no subsampling]\n"
// general options
" -@, --threads INT\n"
" number of BAM/CRAM compression threads [0]\n"
" -? print long help, including note about region specification\n"
" -S ignored (input format is auto-detected)\n");
sam_global_opt_help(fp, "-.O.T");
fprintf(fp, "\n");
if (is_long_help)
fprintf(fp,
"Notes:\n"
"\n"
"1. This command now auto-detects the input format (BAM/CRAM/SAM).\n"
" Further control over the CRAM format can be specified by using the\n"
" --output-fmt-option, e.g. to specify the number of sequences per slice\n"
" and to use avoid reference based compression:\n"
"\n"
"\tsamtools view -C --output-fmt-option seqs_per_slice=5000 \\\n"
"\t --output-fmt-option no_ref -o out.cram in.bam\n"
"\n"
" Options can also be specified as a comma separated list within the\n"
" --output-fmt value too. For example this is equivalent to the above\n"
"\n"
"\tsamtools view --output-fmt cram,seqs_per_slice=5000,no_ref \\\n"
"\t -o out.cram in.bam\n"
"\n"
"2. The file supplied with `-t' is SPACE/TAB delimited with the first\n"
" two fields of each line consisting of the reference name and the\n"
" corresponding sequence length. The `.fai' file generated by \n"
" `samtools faidx' is suitable for use as this file. This may be an\n"
" empty file if reads are unaligned.\n"
"\n"
"3. SAM->BAM conversion: samtools view -bT ref.fa in.sam.gz\n"
"\n"
"4. BAM->SAM conversion: samtools view -h in.bam\n"
"\n"
"5. 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 a sorted and indexed\n"
" alignment (BAM/CRAM) file.\n"
"\n"
"6. Option `-u' is preferred over `-b' when the output is piped to\n"
" another samtools command.\n"
"\n");
return exit_status;
}
static bool split(state_t* state)
{
if (state->unaccounted_file && sam_hdr_write(state->unaccounted_file, state->unaccounted_header) != 0) {
fprintf(pysamerr, "Could not write output file header\n");
return false;
}
size_t i;
for (i = 0; i < state->output_count; i++) {
if (sam_hdr_write(state->rg_output_file[i], state->rg_output_header[i]) != 0) {
fprintf(pysamerr, "Could not write output file header\n");
return false;
}
}
bam1_t* file_read = bam_init1();
// Read the first record
if (sam_read1(state->merged_input_file, state->merged_input_header, file_read) < 0) {
// Nothing more to read? Ignore this file
bam_destroy1(file_read);
file_read = NULL;
}
while (file_read != NULL) {
// Get RG tag from read and look it up in hash to find file to output it to
uint8_t* tag = bam_aux_get(file_read, "RG");
khiter_t iter;
if ( tag != NULL ) {
char* rg = bam_aux2Z(tag);
iter = kh_get_c2i(state->rg_hash, rg);
} else {
iter = kh_end(state->rg_hash);
}
// Write the read out to correct file
if (iter != kh_end(state->rg_hash)) {
// if found write to the appropriate untangled bam
int i = kh_val(state->rg_hash,iter);
sam_write1(state->rg_output_file[i], state->rg_output_header[i], file_read);
} else {
// otherwise write to the unaccounted bam if there is one or fail
if (state->unaccounted_file == NULL) {
if (tag) {
fprintf(pysamerr, "Read \"%s\" with unaccounted for tag \"%s\".\n", bam_get_qname(file_read), bam_aux2Z(tag));
} else {
fprintf(pysamerr, "Read \"%s\" has no RG tag.\n", bam_get_qname(file_read));
}
bam_destroy1(file_read);
return false;
} else {
sam_write1(state->unaccounted_file, state->unaccounted_header, file_read);
}
}
// Replace written read with the next one to process
if (sam_read1(state->merged_input_file, state->merged_input_header, file_read) < 0) {
// Nothing more to read? Ignore this file in future
bam_destroy1(file_read);
file_read = NULL;
}
}
return true;
}
int main(int argc, char *argv[])
{
samFile *in;
char *fn_ref = 0;
int flag = 0, c, clevel = -1, ignore_sam_err = 0;
char moder[8];
bam_hdr_t *h;
bam1_t *b;
htsFile *out;
char modew[8];
int r = 0, exit_code = 0;
while ((c = getopt(argc, argv, "IbDCSl:t:")) >= 0) {
switch (c) {
case 'S': flag |= 1; break;
case 'b': flag |= 2; break;
case 'D': flag |= 4; break;
case 'C': flag |= 8; break;
case 'l': clevel = atoi(optarg); flag |= 2; break;
case 't': fn_ref = optarg; break;
case 'I': ignore_sam_err = 1; break;
}
}
if (argc == optind) {
fprintf(stderr, "Usage: samview [-bSCSI] [-l level] <in.bam>|<in.sam>|<in.cram> [region]\n");
return 1;
}
strcpy(moder, "r");
if (flag&4) strcat(moder, "c");
else if ((flag&1) == 0) strcat(moder, "b");
in = sam_open(argv[optind], moder);
h = sam_hdr_read(in);
h->ignore_sam_err = ignore_sam_err;
b = bam_init1();
strcpy(modew, "w");
if (clevel >= 0 && clevel <= 9) sprintf(modew + 1, "%d", clevel);
if (flag&8) strcat(modew, "c");
else if (flag&2) strcat(modew, "b");
out = hts_open("-", modew);
/* CRAM output */
if (flag & 8) {
// Parse input header and use for CRAM output
out->fp.cram->header = sam_hdr_parse_(h->text, h->l_text);
// Create CRAM references arrays
if (fn_ref)
cram_set_option(out->fp.cram, CRAM_OPT_REFERENCE, fn_ref);
else
// Attempt to fill out a cram->refs[] array from @SQ headers
cram_set_option(out->fp.cram, CRAM_OPT_REFERENCE, NULL);
}
sam_hdr_write(out, h);
if (optind + 1 < argc && !(flag&1)) { // BAM input and has a region
int i;
hts_idx_t *idx;
if ((idx = bam_index_load(argv[optind])) == 0) {
fprintf(stderr, "[E::%s] fail to load the BAM index\n", __func__);
return 1;
}
for (i = optind + 1; i < argc; ++i) {
hts_itr_t *iter;
if ((iter = bam_itr_querys(idx, h, argv[i])) == 0) {
fprintf(stderr, "[E::%s] fail to parse region '%s'\n", __func__, argv[i]);
continue;
}
while ((r = bam_itr_next(in, iter, b)) >= 0) {
if (sam_write1(out, h, b) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
}
}
hts_itr_destroy(iter);
}
hts_idx_destroy(idx);
} else while ((r = sam_read1(in, h, b)) >= 0) {
if (sam_write1(out, h, b) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
}
}
sam_close(out);
if (r < -1) {
fprintf(stderr, "Error parsing input.\n");
exit_code = 1;
}
bam_destroy1(b);
bam_hdr_destroy(h);
sam_close(in);
return exit_code;
}
int main(int argc, char *argv[])
{
samFile *in;
char *fn_ref = 0;
int flag = 0, c, clevel = -1, ignore_sam_err = 0;
char moder[8];
bam_hdr_t *h;
bam1_t *b;
htsFile *out;
char modew[8];
int r = 0, exit_code = 0;
hts_opt *in_opts = NULL, *out_opts = NULL, *last = NULL;
int nreads = 0;
int benchmark = 0;
while ((c = getopt(argc, argv, "IbDCSl:t:i:o:N:B")) >= 0) {
switch (c) {
case 'S': flag |= 1; break;
case 'b': flag |= 2; break;
case 'D': flag |= 4; break;
case 'C': flag |= 8; break;
case 'B': benchmark = 1; break;
case 'l': clevel = atoi(optarg); flag |= 2; break;
case 't': fn_ref = optarg; break;
case 'I': ignore_sam_err = 1; break;
case 'i': if (add_option(&in_opts, optarg)) return 1; break;
case 'o': if (add_option(&out_opts, optarg)) return 1; break;
case 'N': nreads = atoi(optarg);
}
}
if (argc == optind) {
fprintf(stderr, "Usage: samview [-bSCSIB] [-N num_reads] [-l level] [-o option=value] <in.bam>|<in.sam>|<in.cram> [region]\n");
return 1;
}
strcpy(moder, "r");
if (flag&4) strcat(moder, "c");
else if ((flag&1) == 0) strcat(moder, "b");
in = sam_open(argv[optind], moder);
if (in == NULL) {
fprintf(stderr, "Error opening \"%s\"\n", argv[optind]);
return EXIT_FAILURE;
}
h = sam_hdr_read(in);
h->ignore_sam_err = ignore_sam_err;
b = bam_init1();
strcpy(modew, "w");
if (clevel >= 0 && clevel <= 9) sprintf(modew + 1, "%d", clevel);
if (flag&8) strcat(modew, "c");
else if (flag&2) strcat(modew, "b");
out = hts_open("-", modew);
if (out == NULL) {
fprintf(stderr, "Error opening standard output\n");
return EXIT_FAILURE;
}
/* CRAM output */
if (flag & 8) {
int ret;
// Parse input header and use for CRAM output
out->fp.cram->header = sam_hdr_parse_(h->text, h->l_text);
// Create CRAM references arrays
if (fn_ref)
ret = cram_set_option(out->fp.cram, CRAM_OPT_REFERENCE, fn_ref);
else
// Attempt to fill out a cram->refs[] array from @SQ headers
ret = cram_set_option(out->fp.cram, CRAM_OPT_REFERENCE, NULL);
if (ret != 0)
return EXIT_FAILURE;
}
// Process any options; currently cram only.
for (; in_opts; in_opts = (last=in_opts)->next, free(last)) {
hts_set_opt(in, in_opts->opt, in_opts->val);
if (in_opts->opt == CRAM_OPT_REFERENCE)
if (hts_set_opt(out, in_opts->opt, in_opts->val) != 0)
return EXIT_FAILURE;
}
for (; out_opts; out_opts = (last=out_opts)->next, free(last))
if (hts_set_opt(out, out_opts->opt, out_opts->val) != 0)
return EXIT_FAILURE;
if (!benchmark)
sam_hdr_write(out, h);
if (optind + 1 < argc && !(flag&1)) { // BAM input and has a region
int i;
hts_idx_t *idx;
if ((idx = sam_index_load(in, argv[optind])) == 0) {
fprintf(stderr, "[E::%s] fail to load the BAM index\n", __func__);
return 1;
}
for (i = optind + 1; i < argc; ++i) {
hts_itr_t *iter;
if ((iter = sam_itr_querys(idx, h, argv[i])) == 0) {
fprintf(stderr, "[E::%s] fail to parse region '%s'\n", __func__, argv[i]);
continue;
}
while ((r = sam_itr_next(in, iter, b)) >= 0) {
if (!benchmark && sam_write1(out, h, b) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
}
if (nreads && --nreads == 0)
break;
}
hts_itr_destroy(iter);
}
hts_idx_destroy(idx);
} else while ((r = sam_read1(in, h, b)) >= 0) {
if (!benchmark && sam_write1(out, h, b) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
}
if (nreads && --nreads == 0)
break;
}
if (r < -1) {
fprintf(stderr, "Error parsing input.\n");
exit_code = 1;
}
r = sam_close(out);
if (r < 0) {
fprintf(stderr, "Error closing output.\n");
exit_code = 1;
}
bam_destroy1(b);
bam_hdr_destroy(h);
r = sam_close(in);
if (r < 0) {
fprintf(stderr, "Error closing input.\n");
exit_code = 1;
}
return exit_code;
}
int bam_fillmd(int argc, char *argv[])
{
int c, flt_flag, tid = -2, ret, len, is_bam_out, is_uncompressed, max_nm, is_realn, capQ, baq_flag;
samFile *fp = NULL, *fpout = NULL;
bam_hdr_t *header = NULL;
faidx_t *fai = NULL;
char *ref = NULL, mode_w[8], *ref_file;
bam1_t *b = NULL;
sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
static const struct option lopts[] = {
SAM_OPT_GLOBAL_OPTIONS('-', 0, 0, 0, 0),
{ NULL, 0, NULL, 0 }
};
flt_flag = UPDATE_NM | UPDATE_MD;
is_bam_out = is_uncompressed = is_realn = max_nm = capQ = baq_flag = 0;
strcpy(mode_w, "w");
while ((c = getopt_long(argc, argv, "EqreuNhbSC:n:Ad", lopts, NULL)) >= 0) {
switch (c) {
case 'r':
is_realn = 1;
break;
case 'e':
flt_flag |= USE_EQUAL;
break;
case 'd':
flt_flag |= DROP_TAG;
break;
case 'q':
flt_flag |= BIN_QUAL;
break;
case 'h':
flt_flag |= HASH_QNM;
break;
case 'N':
flt_flag &= ~(UPDATE_MD|UPDATE_NM);
break;
case 'b':
is_bam_out = 1;
break;
case 'u':
is_uncompressed = is_bam_out = 1;
break;
case 'S':
break;
case 'n':
max_nm = atoi(optarg);
break;
case 'C':
capQ = atoi(optarg);
break;
case 'A':
baq_flag |= 1;
break;
case 'E':
baq_flag |= 2;
break;
default:
if (parse_sam_global_opt(c, optarg, lopts, &ga) == 0) break;
fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n\n", c);
/* else fall-through */
case '?':
return calmd_usage();
}
}
if (is_bam_out) strcat(mode_w, "b");
else strcat(mode_w, "h");
if (is_uncompressed) strcat(mode_w, "0");
if (optind + (ga.reference == NULL) >= argc)
return calmd_usage();
fp = sam_open_format(argv[optind], "r", &ga.in);
if (fp == NULL) {
print_error_errno("calmd", "Failed to open input file '%s'", argv[optind]);
return 1;
}
header = sam_hdr_read(fp);
if (header == NULL || header->n_targets == 0) {
fprintf(stderr, "[bam_fillmd] input SAM does not have header. Abort!\n");
goto fail;
}
fpout = sam_open_format("-", mode_w, &ga.out);
if (fpout == NULL) {
print_error_errno("calmd", "Failed to open output");
goto fail;
}
if (sam_hdr_write(fpout, header) < 0) {
print_error_errno("calmd", "Failed to write sam header");
goto fail;
}
ref_file = argc > optind + 1 ? argv[optind+1] : ga.reference;
fai = fai_load(ref_file);
if (!fai) {
print_error_errno("calmd", "Failed to open reference file '%s'", ref_file);
goto fail;
}
b = bam_init1();
if (!b) {
fprintf(stderr, "[bam_fillmd] Failed to allocate bam struct\n");
goto fail;
}
while ((ret = sam_read1(fp, header, b)) >= 0) {
if (b->core.tid >= 0) {
if (tid != b->core.tid) {
free(ref);
ref = fai_fetch(fai, header->target_name[b->core.tid], &len);
tid = b->core.tid;
if (ref == 0) { // FIXME: Should this always be fatal?
fprintf(stderr, "[bam_fillmd] fail to find sequence '%s' in the reference.\n",
header->target_name[tid]);
if (is_realn || capQ > 10) goto fail; // Would otherwise crash
}
}
if (is_realn) sam_prob_realn(b, ref, len, baq_flag);
if (capQ > 10) {
int q = sam_cap_mapq(b, ref, len, capQ);
if (b->core.qual > q) b->core.qual = q;
}
if (ref) bam_fillmd1_core(b, ref, len, flt_flag, max_nm);
}
if (sam_write1(fpout, header, b) < 0) {
print_error_errno("calmd", "failed to write to output file");
goto fail;
}
}
if (ret < -1) {
fprintf(stderr, "[bam_fillmd] Error reading input.\n");
goto fail;
}
bam_destroy1(b);
bam_hdr_destroy(header);
free(ref);
fai_destroy(fai);
sam_close(fp);
if (sam_close(fpout) < 0) {
fprintf(stderr, "[bam_fillmd] error when closing output file\n");
return 1;
}
return 0;
fail:
free(ref);
if (b) bam_destroy1(b);
if (header) bam_hdr_destroy(header);
if (fai) fai_destroy(fai);
if (fp) sam_close(fp);
if (fpout) sam_close(fpout);
return 1;
}
int main_pad2unpad(int argc, char *argv[])
{
samFile *in = 0, *out = 0;
bam_hdr_t *h = 0, *h_fix = 0;
faidx_t *fai = 0;
int c, compress_level = -1, is_long_help = 0;
char in_mode[5], out_mode[6], *fn_out = 0, *fn_list = 0;
int ret=0;
sam_global_args ga = SAM_GLOBAL_ARGS_INIT;
static const struct option lopts[] = {
SAM_OPT_GLOBAL_OPTIONS('-', 0, 0, 0, 'T'),
{ NULL, 0, NULL, 0 }
};
/* parse command-line options */
strcpy(in_mode, "r"); strcpy(out_mode, "w");
while ((c = getopt_long(argc, argv, "SCso:u1T:?", lopts, NULL)) >= 0) {
switch (c) {
case 'S': break;
case 'C': hts_parse_format(&ga.out, "cram"); break;
case 's': assert(compress_level == -1); hts_parse_format(&ga.out, "sam"); break;
case 'o': fn_out = strdup(optarg); break;
case 'u':
compress_level = 0;
if (ga.out.format == unknown_format)
hts_parse_format(&ga.out, "bam");
break;
case '1':
compress_level = 1;
if (ga.out.format == unknown_format)
hts_parse_format(&ga.out, "bam");
break;
case '?': is_long_help = 1; break;
default: if (parse_sam_global_opt(c, optarg, lopts, &ga) == 0) break;
fprintf(stderr, "[bam_fillmd] unrecognized option '-%c'\n\n", c);
return usage(is_long_help);
}
}
if (argc == optind) return usage(is_long_help);
strcat(out_mode, "h");
if (compress_level >= 0) {
char tmp[2];
tmp[0] = compress_level + '0'; tmp[1] = '\0';
strcat(out_mode, tmp);
}
// Load FASTA reference (also needed for SAM -> BAM if missing header)
if (ga.reference) {
fn_list = samfaipath(ga.reference);
fai = fai_load(ga.reference);
}
// open file handlers
if ((in = sam_open_format(argv[optind], in_mode, &ga.in)) == 0) {
fprintf(stderr, "[depad] failed to open \"%s\" for reading.\n", argv[optind]);
ret = 1;
goto depad_end;
}
if (fn_list && hts_set_fai_filename(in, fn_list) != 0) {
fprintf(stderr, "[depad] failed to load reference file \"%s\".\n", fn_list);
ret = 1;
goto depad_end;
}
if ((h = sam_hdr_read(in)) == 0) {
fprintf(stderr, "[depad] failed to read the header from \"%s\".\n", argv[optind]);
ret = 1;
goto depad_end;
}
if (fai) {
h_fix = fix_header(h, fai);
} else {
fprintf(stderr, "[depad] Warning - reference lengths will not be corrected without FASTA reference\n");
h_fix = h;
}
char wmode[2];
strcat(out_mode, sam_open_mode(wmode, fn_out, NULL)==0 ? wmode : "b");
if ((out = sam_open_format(fn_out? fn_out : "-", out_mode, &ga.out)) == 0) {
fprintf(stderr, "[depad] failed to open \"%s\" for writing.\n", fn_out? fn_out : "standard output");
ret = 1;
goto depad_end;
}
// Reference-based CRAM won't work unless we also create a new reference.
// We could embed this, but for now we take the easy option.
if (ga.out.format == cram)
hts_set_opt(out, CRAM_OPT_NO_REF, 1);
if (sam_hdr_write(out, h_fix) != 0) {
fprintf(stderr, "[depad] failed to write header.\n");
ret = 1;
goto depad_end;
}
// Do the depad
ret = bam_pad2unpad(in, out, h, fai);
depad_end:
// close files, free and return
if (fai) fai_destroy(fai);
if (h) bam_hdr_destroy(h);
sam_close(in);
sam_close(out);
free(fn_list); free(fn_out);
return ret;
}
/*
* CRAM files don't store the RG:Z:ID per read in the aux field.
* Instead they have a numerical data series (RG) to point each read
* back to the Nth @RG line in the file. This means that we may need
* to edit the RG data series (if the files were produced from
* "samtools split" for example).
*
* The encoding method is stored in the compression header. Typical
* examples:
*
* RG => EXTERNAL {18} # Block content-id 18 holds RG values
* # as a series of ITF8 encoded values
*
* RG => HUFFMAN {1, 255, 255, 255, 255, 255, 1, 0}
* # One RG value #-1. (No RG)
*
* RG => HUFFMAN {1, 0, 1, 0} # One RG value #0 (always first RG)
*
* RG => HUFFMAN {2, 0, 1, 2, 1, 1}
* # Two RG values, #0 and #1, written
* # to the CORE block and possibly
* # mixed with other data series.
*
* A single value can (but may not be) implemented as a zero bit
* huffman code. In this situation we can change the meta-data in the
* compression header to renumber an RG value..
*/
int cram_cat(int nfn, char * const *fn, const bam_hdr_t *h, const char* outcram)
{
samFile *out;
cram_fd *out_c;
int i, vers_maj, vers_min;
khash_s2i *rg2id = NULL;
bam_hdr_t *new_h = NULL;
/* Check consistent versioning and compatible headers */
if (!(new_h = cram_cat_check_hdr(nfn, fn, h, &rg2id, &vers_maj, &vers_min)))
return -1;
/* Open the file with cram_vers */
char vers[100];
sprintf(vers, "%d.%d", vers_maj, vers_min);
out = sam_open(outcram, "wc");
if (out == 0) {
fprintf(stderr, "[%s] ERROR: fail to open output file '%s'.\n", __func__, outcram);
return 1;
}
out_c = out->fp.cram;
cram_set_option(out_c, CRAM_OPT_VERSION, vers);
//fprintf(stderr, "Creating cram vers %s\n", vers);
cram_fd_set_header(out_c, sam_hdr_parse_(new_h->text, new_h->l_text)); // needed?
sam_hdr_write(out, new_h);
for (i = 0; i < nfn; ++i) {
samFile *in;
cram_fd *in_c;
cram_container *c;
bam_hdr_t *old;
int new_rg = -1;
in = sam_open(fn[i], "rc");
if (in == 0) {
fprintf(stderr, "[%s] ERROR: fail to open file '%s'.\n", __func__, fn[i]);
return -1;
}
in_c = in->fp.cram;
old = sam_hdr_read(in);
khash_s2i *rg2id_in = hash_rg(old);
// Compute RG mapping if suitable for changing.
if (rg2id_in->n_id == 1) {
int _;
new_rg = hash_s2i_inc(rg2id, rg2id_in->id[0], NULL, &_);
} else {
new_rg = 0;
}
hash_s2i_free(rg2id_in);
// Copy contains and blocks within them
while ((c = cram_read_container(in_c))) {
cram_block *blk;
if (cram_container_is_empty(in_c)) {
if (cram_write_container(out_c, c) != 0)
return -1;
// Container compression header
if (!(blk = cram_read_block(in_c)))
return -1;
if (cram_write_block(out_c, blk) != 0) {
cram_free_block(blk);
return -1;
}
cram_free_block(blk);
cram_free_container(c);
continue;
}
// If we have just one RG key and new_rg != 0 then
// we need to edit the compression header. IF WE CAN.
if (new_rg) {
int zero = 0;
//fprintf(stderr, "Transcode RG %d to %d\n", 0, new_rg);
cram_transcode_rg(in_c, out_c, c, 1, &zero, &new_rg);
} else {
int32_t num_slices;
// Not switching rg so do the usual read/write loop
if (cram_write_container(out_c, c) != 0)
return -1;
// Container compression header
if (!(blk = cram_read_block(in_c)))
return -1;
if (cram_write_block(out_c, blk) != 0) {
cram_free_block(blk);
return -1;
}
cram_free_block(blk);
// Container num_blocks can be invalid, due to a bug.
// Instead we iterate in slice context instead.
(void)cram_container_get_landmarks(c, &num_slices);
cram_copy_slice(in_c, out_c, num_slices);
}
cram_free_container(c);
}
bam_hdr_destroy(old);
sam_close(in);
}
sam_close(out);
hash_s2i_free(rg2id);
bam_hdr_destroy(new_h);
return 0;
}
int main(int argc, char **argv)
{
if (argc < 4)
errx(1,
"usage\t:%s <bam> <split out> <discord out> (optional #threads)",
argv[0]);
char *bam_file_name = argv[1];
char *split_file_name = argv[2];
char *disc_file_name = argv[3];
int threads = 2;
if (argc == 5) {
threads = atoi(argv[4]);
}
samFile *disc = sam_open(disc_file_name, "wb");
samFile *split = sam_open(split_file_name, "wb");
samFile *in = sam_open(bam_file_name, "rb");
if(in == NULL)
errx(1, "Unable to open BAM/SAM file.");
// TODO: handle cram.
if (threads > 1) {
bgzf_mt(in->fp.bgzf, threads, 256);
}
hts_idx_t *idx = sam_index_load(in, bam_file_name);
if(idx == NULL)
errx(1,"Unable to open BAM/SAM index.");
bam_hdr_t *hdr = sam_hdr_read(in);
int r = sam_hdr_write(disc, hdr);
r = sam_hdr_write(split, hdr);
bam1_t *aln = bam_init1();
int ret;
while(ret = sam_read1(in, hdr, aln) >= 0) {
if (((aln->core.flag) & 1294) == 0)
r = sam_write1(disc, hdr, aln);
uint8_t *sa = bam_aux_get(aln, "SA");
if (sa != 0) {
char *sa_tag = strdup(bam_aux2Z(sa));
if ( count_tags(sa_tag) == 1) {
char *chrm, strand, *cigar;
uint32_t pos;
split_sa_tag(sa_tag,
&chrm,
&pos,
&strand,
&cigar);
struct line sa, al;
calcOffsets(cigar,
pos,
strand,
&sa);
sa.chrm = chrm;
sa.strand = strand;
calcAlnOffsets(bam_get_cigar(aln),
aln->core.n_cigar,
aln->core.pos,
bam_is_rev(aln) ? '-' : '+',
&al);
al.chrm = hdr->target_name[aln->core.tid];
al.strand = bam_is_rev(aln) ? '-' : '+';
struct line *left = &al, *right = &sa;
if (left->SQO > right->SQO) {
left = &sa;
right = &al;
}
int overlap = MAX(1 + MIN(left->EQO, right->EQO) -
MAX(left->SQO, right->SQO), 0);
int alen1 = 1 + left->EQO - left->SQO;
int alen2 = 1 + right->EQO - right->SQO;
int mno = MIN(alen1-overlap, alen2-overlap);
if (mno < MIN_NON_OVERLAP)
continue;
if ( (strcmp(left->chrm, right->chrm) == 0) &&
(left->strand == right->strand) ) {
int leftDiag, rightDiag, insSize;
if (left->strand == '-') {
leftDiag = left->rapos - left->sclip;
rightDiag = (right->rapos + right->raLen) -
(right->sclip + right->qaLen);
insSize = rightDiag - leftDiag;
} else {
leftDiag = (left->rapos + left->raLen) -
(left->sclip + left->qaLen);
rightDiag = right->rapos - right->sclip;
insSize = leftDiag - rightDiag;
}
int desert = right->SQO - left->EQO - 1;
if ((abs(insSize) < MIN_INDEL_SIZE) ||
((desert > 0) && (
(desert - (int)MAX(0, insSize)) >
MAX_UNMAPPED_BASES)))
continue;
}
char *qname = bam_get_qname(aln);
if ((aln->core.flag & 64) == 64)
qname[0] = 'A';
else
qname[0] = 'B';
r = sam_write1(split, hdr, aln);
}
free(sa_tag);
}
}
bam_destroy1(aln);
hts_idx_destroy(idx);
bam_hdr_destroy(hdr);
sam_close(in);
sam_close(disc);
sam_close(split);
if(ret < -1) {
errx(1, "lumpy_filter: error reading bam: %s\n", bam_file_name);
}
}
// currently, this function ONLY works if each read has one hit
static void bam_mating_core(samFile* in, samFile* out, int remove_reads, int proper_pair_check, int add_ct)
{
bam_hdr_t *header;
bam1_t *b[2];
int curr, has_prev, pre_end = 0, cur_end = 0;
kstring_t str;
str.l = str.m = 0; str.s = 0;
header = sam_hdr_read(in);
if (header == NULL) {
fprintf(stderr, "[bam_mating_core] ERROR: Couldn't read header\n");
exit(1);
}
// Accept unknown, unsorted, or queryname sort order, but error on coordinate sorted.
if ((header->l_text > 3) && (strncmp(header->text, "@HD", 3) == 0)) {
char *p, *q;
p = strstr(header->text, "\tSO:coordinate");
q = strchr(header->text, '\n');
// Looking for SO:coordinate within the @HD line only
// (e.g. must ignore in a @CO comment line later in header)
if ((p != 0) && (p < q)) {
fprintf(stderr, "[bam_mating_core] ERROR: Coordinate sorted, require grouped/sorted by queryname.\n");
exit(1);
}
}
sam_hdr_write(out, header);
b[0] = bam_init1();
b[1] = bam_init1();
curr = 0; has_prev = 0;
while (sam_read1(in, header, b[curr]) >= 0) {
bam1_t *cur = b[curr], *pre = b[1-curr];
if (cur->core.flag & BAM_FSECONDARY)
{
if ( !remove_reads ) sam_write1(out, header, cur);
continue; // skip secondary alignments
}
if (cur->core.flag & BAM_FSUPPLEMENTARY)
{
sam_write1(out, header, cur);
continue; // pass supplementary alignments through unchanged (TODO:make them match read they came from)
}
if (cur->core.tid < 0 || cur->core.pos < 0) // If unmapped set the flag
{
cur->core.flag |= BAM_FUNMAP;
}
if ((cur->core.flag&BAM_FUNMAP) == 0) // If mapped calculate end
{
cur_end = bam_endpos(cur);
// Check cur_end isn't past the end of the contig we're on, if it is set the UNMAP'd flag
if (cur_end > (int)header->target_len[cur->core.tid]) cur->core.flag |= BAM_FUNMAP;
}
if (has_prev) { // do we have a pair of reads to examine?
if (strcmp(bam_get_qname(cur), bam_get_qname(pre)) == 0) { // identical pair name
pre->core.flag |= BAM_FPAIRED;
cur->core.flag |= BAM_FPAIRED;
sync_mate(pre, cur);
if (pre->core.tid == cur->core.tid && !(cur->core.flag&(BAM_FUNMAP|BAM_FMUNMAP))
&& !(pre->core.flag&(BAM_FUNMAP|BAM_FMUNMAP))) // if safe 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 (add_ct) bam_template_cigar(pre, cur, &str);
// TODO: Add code to properly check if read is in a proper pair based on ISIZE distribution
if (proper_pair_check && !plausibly_properly_paired(pre,cur)) {
pre->core.flag &= ~BAM_FPROPER_PAIR;
cur->core.flag &= ~BAM_FPROPER_PAIR;
}
// Write out result
if ( !remove_reads ) {
sam_write1(out, header, pre);
sam_write1(out, header, cur);
} else {
// If we have to remove reads make sure we do it in a way that doesn't create orphans with bad flags
if(pre->core.flag&BAM_FUNMAP) cur->core.flag &= ~(BAM_FPAIRED|BAM_FMREVERSE|BAM_FPROPER_PAIR);
if(cur->core.flag&BAM_FUNMAP) pre->core.flag &= ~(BAM_FPAIRED|BAM_FMREVERSE|BAM_FPROPER_PAIR);
if(!(pre->core.flag&BAM_FUNMAP)) sam_write1(out, header, pre);
if(!(cur->core.flag&BAM_FUNMAP)) sam_write1(out, header, cur);
}
has_prev = 0;
} else { // unpaired? clear bad info and write it out
if (pre->core.tid < 0 || pre->core.pos < 0 || pre->core.flag&BAM_FUNMAP) { // If unmapped
pre->core.flag |= BAM_FUNMAP;
pre->core.tid = -1;
pre->core.pos = -1;
}
pre->core.mtid = -1; pre->core.mpos = -1; pre->core.isize = 0;
pre->core.flag &= ~(BAM_FPAIRED|BAM_FMREVERSE|BAM_FPROPER_PAIR);
if ( !remove_reads || !(pre->core.flag&BAM_FUNMAP) ) sam_write1(out, header, pre);
}
} else has_prev = 1;
curr = 1 - curr;
pre_end = cur_end;
}
if (has_prev && !remove_reads) { // If we still have a BAM in the buffer it must be unpaired
bam1_t *pre = b[1-curr];
if (pre->core.tid < 0 || pre->core.pos < 0 || pre->core.flag&BAM_FUNMAP) { // If unmapped
pre->core.flag |= BAM_FUNMAP;
pre->core.tid = -1;
pre->core.pos = -1;
}
pre->core.mtid = -1; pre->core.mpos = -1; pre->core.isize = 0;
pre->core.flag &= ~(BAM_FPAIRED|BAM_FMREVERSE|BAM_FPROPER_PAIR);
sam_write1(out, header, pre);
}
bam_hdr_destroy(header);
bam_destroy1(b[0]);
bam_destroy1(b[1]);
free(str.s);
}
int do_grep() {
#ifdef DEBUGa
printf("[!]do_grep\n");
#endif
BamInfo_t *pbam;
kh_cstr_t BamID;
khiter_t ki, bami;
kstring_t ks1 = { 0, 0, NULL };
kstring_t ks2 = { 0, 0, NULL };
kstring_t ks3 = { 0, 0, NULL };
kstring_t kstr = { 0, 0, NULL };
//ksprintf(kstr, "%s/%s_grep/", myConfig.WorkDir, myConfig.ProjectID);
//const char *filePrefix = strdup(ks_str(kstr));
//kputs(myConfig.WorkDir,kstr);
samFile *in;
bam_hdr_t *h;
hts_idx_t *idx;
bam1_t *b, *d, *d2, *bR1, *bR2, *bR3;
bR1 = bam_init1(); bR2 = bam_init1(); bR3 = bam_init1();
htsFile *out;
//hts_opt *in_opts = NULL, *out_opts = NULL;
int r = 0, exit_code = 0;
kvec_t(bam1_t) R1, R2, RV;
pierCluster_t *pierCluster;
//samdat_t tmp_samdat;
#ifdef DEBUGa
kstr.l = 0;
ksprintf(&kstr, "%s/%s_grep/Greped.dump", myConfig.WorkDir, myConfig.ProjectID);
FILE *fsdump = fopen(ks_str(&kstr),"w");
#endif
kstr.l = 0;
ksprintf(&kstr, "%s/%s_grep/Greped.ini", myConfig.WorkDir, myConfig.ProjectID);
FILE *fs = fopen(ks_str(&kstr),"w");
uint32_t blockid = 0;
for (bami = kh_begin(bamNFOp); bami != kh_end(bamNFOp); ++bami) {
//printf(">[%d]:\n",bami);
if (kh_exist(bamNFOp, bami)) {
kv_init(R1); kv_init(R2); kv_init(RV);
//tmp_samdat = (const samdat_t){ 0 };
//memset(&tmp_samdat,0,sizeof(samdat_t));
//printf("-[%d]:\n",bami);
BamID = kh_key(bamNFOp, bami);
pbam = &kh_value(bamNFOp, bami);
fprintf(stderr, "%u [%s]=%s\t%u %u\n",bami,BamID,pbam->fileName,pbam->insertSize,pbam->SD);
in = sam_open(pbam->fileName, "r");
if (in == NULL) {
fprintf(stderr, "[x]Error opening \"%s\"\n", pbam->fileName);
return EXIT_FAILURE;
}
h = sam_hdr_read(in);
kstr.l = 0;
ksprintf(&kstr, "%s/%s_grep/%s.bam", myConfig.WorkDir, myConfig.ProjectID, BamID);
out = hts_open(ks_str(&kstr), "wb");
if (out == NULL) {
fprintf(stderr, "[x]Error opening [%s]\n",ks_str(&kstr));
return EXIT_FAILURE;
}
if (sam_hdr_write(out, h) < 0) {
fprintf(stderr, "[!]Error writing output header.\n");
exit_code = 1;
}
int8_t *ChrIsHum;
if (h == NULL) {
fprintf(stderr, "[x]Couldn't read header for \"%s\"\n", pbam->fileName);
return EXIT_FAILURE;
} else {
ChrIsHum = (int8_t *) malloc(h->n_targets * sizeof(int8_t));
for (int32_t i=0; i < h->n_targets; ++i) {
//ChrIsHum[i] = -1;
ki = kh_get(chrNFO, chrNFOp, h->target_name[i]);
if (ki == kh_end(chrNFOp)) {
errx(4,"[x]Cannot find ChrID for [%s] !",h->target_name[i]);
} else {
ChrInfo_t * tmp = &kh_value(chrNFOp, ki);
ChrIsHum[i] = tmp->isHum;
//printf(">>> %d Chr:%s %d\n",i,h->target_name[i],ChrIsHum[i]);
}
}
}
h->ignore_sam_err = 0;
b = bam_init1();
d = bam_init1();
d2 = bam_init1();
if ((idx = sam_index_load(in, pbam->fileName)) == 0) {
fprintf(stderr, "[E::%s] fail to load the BAM index\n", __func__);
return 1;
}
pierCluster = sam_plp_init();
while ((r = sam_read1(in, h, b)) >= 0) {
int8_t flag = false;
const bam1_core_t *c = &b->core;
if (c->qual < myConfig.minBamQual) {
continue;
}
if (c->n_cigar) {
uint32_t *cigar = bam_get_cigar(b);
int i = c->n_cigar; --i;
if ( (bam_cigar_opchr(cigar[0])=='S' && bam_cigar_oplen(cigar[0]) >= myConfig.minGrepSlen) ||
(bam_cigar_opchr(cigar[i])=='S' && bam_cigar_oplen(cigar[i]) >= myConfig.minGrepSlen) ) {
flag = true;
}
/* We only need /\d+S/ on both terminal, NOT inside.
for (int i = 0; i < c->n_cigar; ++i) {
if (bam_cigar_opchr(cigar[i])=='S') { // soft clipping
if ( bam_cigar_oplen(cigar[i]) >= myConfig.minGrepSlen ) {
flag = true;
}
}
}
*/
}
if (flag && ChrIsHum[c->tid]) { // Now, skip Virus items.
//bam_copy1(bR1, b);
flag = 0; // recycle
//int enoughMapQ = 0;
//kstring_t ks = { 0, 0, NULL };
/*if (sam_format1(h, b, &ks1) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
} else */if ((c->mtid == c->tid && ChrIsHum[c->tid]) || (ChrIsHum[c->tid] ^ ChrIsHum[c->mtid])) { // Only grep those mapped on same Human ChrID, or diff species/一方在病毒的情况.
//printf(">[%s]\n",ks_str(&ks1));
flag |= 1;
//tmp_samdat.b = bam_dup1(b);
//kv_push(samdat_t,R1,tmp_samdat);
/*if (checkMapQ(ChrIsHum, b, true)) {
++enoughMapQ;
}*/
}
if (getPairedSam(in, idx, b, d) != 0) {
flag &= ~1;
continue;
} else {
flag |= 2;
/*if (checkMapQ(ChrIsHum, d, false)) {
++enoughMapQ;
}*/
if (c->flag & BAM_FSECONDARY) {
if (getPairedSam(in, idx, d, d2) == 0) {
//sam_format1(h, d2, &ks3);
flag |= 4;
/*if (checkMapQ(ChrIsHum, d2, false)) {
++enoughMapQ;
}*/
}
}
}
/*
对于 BAM_FSECONDARY(256) 的 Read，跳两次 与 读 SA 项，效果一样。
>[sf95_Ref_48245009_48245108_48245208_Vir_-_2000_2044_R_100_90 353 chr2 13996555 0 50S40M chr18 48245109 0ACACAACAATGTTCCGGAGACTCTAAGGCCTCCCGATACAGAGCAGAGGCCACACACACACACACCATGGAATACTATTCAGCCAAAAAA CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC NM:i:0 MD:Z:40 AS:i:40 XS:i:40 RG:Z:Fsimout_mB SA:Z:rgi|59585|emb|X04615.1|,2000,-,40S46M4S,60,0; YC:Z:CT YD:Z:f]
-[sf95_Ref_48245009_48245108_48245208_Vir_-_2000_2044_R_100_90 177 chr18 48245109 9 40S50M gi|59585|emb|X04615.1|2000 0 GTTCCGGAGACTCTAAGGCCTCCCGATACAGAGCAGAGGCCACACACACACACACCATGGAATACTATTCAGCCAAAAAAAGGAATTCAA CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC NM:i:0 MD:Z:50 AS:i:50 XS:i:46 RG:Z:Fsimout_mB SA:Z:rgi|59585|emb|X04615.1|,2000,+,50S40M,9,0; YC:Z:GA YD:Z:f]
+[sf95_Ref_48245009_48245108_48245208_Vir_-_2000_2044_R_100_90 113 gi|59585|emb|X04615.1| 2000 60 40S46M4S chr18 48245109 0 TTTTTTGGCTGAATAGTATTCCATGGTGTGTGTGTGTGTGGCCTCTGCTCTGTATCGGGAGGCCTTAGAGTCTCCGGAACATTGTTGTGT CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC NM:i:0 MD:Z:46 AS:i:46 XS:i:27 RG:Z:Fsimout_mB SA:Z:fchr2,13996555,+,50S40M,0,0; YC:Z:CT YD:Z:r]
*/
/*if (sam_format1(h, d, &ks2) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
}*/
if (((flag & 3) == 3) /*&& enoughMapQ >= myConfig.samples*/) {
/*printf(">%d[%s]\n",checkMapQ(ChrIsHum, b, true),ks_str(&ks1));
printf("-%d[%s]\n",checkMapQ(ChrIsHum, d, false),ks_str(&ks2));
if (flag & 4) {
printf("+%d[%s]\n",checkMapQ(ChrIsHum, d2, false),ks_str(&ks3));
}
printf("<--%d\n",enoughMapQ);*/
bam_aux_append(b, "Zd", 'Z', 2, (uint8_t*)"H");
if (sam_plp_push(ChrIsHum, pierCluster, b) == 0) {
//printf("--HumRange=%s:%d-%d\n", h->target_name[(pierCluster->HumanRange).tid], (pierCluster->HumanRange).pos, (pierCluster->HumanRange).endpos);
if ((!ChrIsHum[(d->core).tid]) && (flag & 2)) {
bam_aux_append(d, "Zd", 'Z', 2, (uint8_t*)"V");
sam_plp_push(ChrIsHum, pierCluster, d);
}
if ((!ChrIsHum[(d2->core).tid]) && (flag & 4)) {
bam_aux_append(d2, "Zd", 'Z', 2, (uint8_t*)"V");
sam_plp_push(ChrIsHum, pierCluster, d2);
}
} else {
++blockid;
//print
#ifdef DEBUGa
fprintf(fsdump,"[%u %s]\nHumRange=%s:%d-%d\n", blockid, BamID, h->target_name[(pierCluster->HumanRange).tid], (pierCluster->HumanRange).pos, (pierCluster->HumanRange).endpos);
fprintf(fsdump,"VirRange=%s:%d-%d\n", h->target_name[(pierCluster->VirusRange).tid], (pierCluster->VirusRange).pos, (pierCluster->VirusRange).endpos);
#endif
fprintf(fs,"[%u]\nBamID=%s\nHumRange=%s:%d-%d\n",blockid, BamID,
h->target_name[(pierCluster->HumanRange).tid], (pierCluster->HumanRange).pos, (pierCluster->HumanRange).endpos);
if ( (pierCluster->VirusRange).pos == 0 && (pierCluster->VirusRange).endpos == 0 ) {
fprintf(fs,"VirRange=NA\n");
} else {
fprintf(fs,"VirRange=%s:%d-%d\n", h->target_name[(pierCluster->VirusRange).tid],
(pierCluster->VirusRange).pos, (pierCluster->VirusRange).endpos);
}
for (size_t i=0; i<kv_size(pierCluster->Reads);++i) {
bam1_t *bi = kv_A(pierCluster->Reads, i);
bam_aux_append(bi, "Zc", 'i', sizeof(uint32_t), (uint8_t*)&blockid);
#ifdef DEBUGa
if (sam_format1(h, bi, &ks1) < 0) {
fprintf(stderr, "Error writing output.\n");
exit_code = 1;
break;
} else {
fprintf(fsdump,"%s\n",ks1.s);
}
#endif
if (sam_write1(out, h, bi) < 0) {
fprintf(stderr, "[x]Error writing output.\n");
exit_code = 1;
break;
}
}
#ifdef DEBUGa
fprintf(fsdump,"\n");
printf("HumRange=%s:%d-%d\n", h->target_name[(pierCluster->HumanRange).tid], (pierCluster->HumanRange).pos, (pierCluster->HumanRange).endpos);
//fflush(fs);
#endif
sam_plp_dectroy(pierCluster);
pierCluster = sam_plp_init();
}
}
}
//char *qname = bam_get_qname(b);
}
r = sam_close(out); // stdout can only be closed once
if (r < 0) {
fprintf(stderr, "Error closing output.\n");
exit_code = 1;
}
hts_idx_destroy(idx);
bam_destroy1(b);
bam_destroy1(d);
bam_destroy1(d2);
bam_hdr_destroy(h);
r = sam_close(in);
free(ChrIsHum);
#ifdef DEBUGa
//fflush(NULL);
//pressAnyKey();
#endif
sam_plp_dectroy(pierCluster);
//printf("<[%d]:\n",bami);
}
}
#ifdef DEBUGa
fclose(fsdump);
#endif
fclose(fs);
getPairedSam(NULL, NULL, NULL, NULL); // sam_close(fp2);
//printf("---[%d]---\n",exit_code);
bam_destroy1(bR1); bam_destroy1(bR2); bam_destroy1(bR3);
ks_release(&ks1);
ks_release(&ks2);
ks_release(&ks3);
ks_release(&kstr);
//free((char*)filePrefix);
return exit_code;
}
int main_samview(int argc, char *argv[])
{
samFile *in;
char *fn_ref = 0;
int flag = 0, c, clevel = -1, ignore_sam_err = 0;
char moder[8];
bam_hdr_t *h;
bam1_t *b;
while ((c = getopt(argc, argv, "IbSl:t:")) >= 0) {
switch (c) {
case 'S': flag |= 1; break;
case 'b': flag |= 2; break;
case 'l': clevel = atoi(optarg); flag |= 2; break;
case 't': fn_ref = optarg; break;
case 'I': ignore_sam_err = 1; break;
}
}
if (argc == optind) {
fprintf(stderr, "Usage: samview [-bSI] [-l level] <in.bam>|<in.sam> [region]\n");
return 1;
}
strcpy(moder, "r");
if ((flag&1) == 0) strcat(moder, "b");
in = sam_open(argv[optind], moder, fn_ref);
h = sam_hdr_read(in);
h->ignore_sam_err = ignore_sam_err;
b = bam_init1();
if ((flag&4) == 0) { // SAM/BAM output
htsFile *out;
char modew[8];
strcpy(modew, "w");
if (clevel >= 0 && clevel <= 9) sprintf(modew + 1, "%d", clevel);
if (flag&2) strcat(modew, "b");
out = hts_open("-", modew, 0);
sam_hdr_write(out, h);
if (optind + 1 < argc && !(flag&1)) { // BAM input and has a region
int i;
hts_idx_t *idx;
if ((idx = bam_index_load(argv[optind])) == 0) {
fprintf(stderr, "[E::%s] fail to load the BAM index\n", __func__);
return 1;
}
for (i = optind + 1; i < argc; ++i) {
hts_itr_t *iter;
if ((iter = bam_itr_querys(idx, h, argv[i])) == 0) {
fprintf(stderr, "[E::%s] fail to parse region '%s'\n", __func__, argv[i]);
continue;
}
while (bam_itr_next((BGZF*)in->fp, iter, b) >= 0) sam_write1(out, h, b);
hts_itr_destroy(iter);
}
hts_idx_destroy(idx);
} else while (sam_read1(in, h, b) >= 0) sam_write1(out, h, b);
sam_close(out);
}
bam_destroy1(b);
bam_hdr_destroy(h);
sam_close(in);
return 0;
}
int main(int argc, char **argv) {
htsFile *in = NULL;
htsFile *out = NULL;
char *in_name = "-";
char *out_name = "-";
char *ref_name = NULL;
char *ref_seq = NULL;
char modew[8] = "w";
faidx_t *fai = NULL;
bam_hdr_t *hdr = NULL;
bam1_t *rec = NULL;
int c, res, last_ref = -1, ref_len = 0;
int adjust = 0, extended = 0, recalc = 0, flags = 0;
while ((c = getopt(argc, argv, "aef:hi:o:r")) >= 0) {
switch (c) {
case 'a': adjust = 1; break;
case 'e': extended = 1; break;
case 'f': ref_name = optarg; break;
case 'h': usage(argv[0]); return EXIT_SUCCESS;
case 'i': in_name = optarg; break;
case 'o': out_name = optarg; break;
case 'r': recalc = 1; break;
default: usage(argv[0]); return EXIT_FAILURE;
}
}
if (!ref_name) {
usage(argv[0]);
return EXIT_FAILURE;
}
flags = (adjust ? 1 : 0) | (extended ? 2 : 0) | (recalc ? 4 : 0);
fai = fai_load(ref_name);
if (!fai) {
fprintf(stderr, "Couldn't load reference %s\n", ref_name);
goto fail;
}
rec = bam_init1();
if (!rec) {
perror(NULL);
goto fail;
}
in = hts_open(in_name, "r");
if (!in) {
fprintf(stderr, "Couldn't open %s : %s\n", in_name, strerror(errno));
goto fail;
}
hdr = sam_hdr_read(in);
if (!hdr) {
fprintf(stderr, "Couldn't read header for %s\n", in_name);
goto fail;
}
out = hts_open(out_name, modew);
if (!out) {
fprintf(stderr, "Couldn't open %s : %s\n", out_name, strerror(errno));
goto fail;
}
if (sam_hdr_write(out, hdr) < 0) {
fprintf(stderr, "Couldn't write header to %s : %s\n",
out_name, strerror(errno));
goto fail;
}
while ((res = sam_read1(in, hdr, rec)) >= 0) {
if (rec->core.tid >= hdr->n_targets) {
fprintf(stderr, "Invalid BAM reference id %d\n", rec->core.tid);
goto fail;
}
if (last_ref != rec->core.tid && rec->core.tid >= 0) {
free(ref_seq);
ref_seq = faidx_fetch_seq(fai, hdr->target_name[rec->core.tid],
0, INT_MAX, &ref_len);
if (!ref_seq) {
fprintf(stderr, "Couldn't get reference %s\n",
hdr->target_name[rec->core.tid]);
goto fail;
}
last_ref = rec->core.tid;
}
if (rec->core.tid >= 0) {
res = sam_prob_realn(rec, ref_seq, ref_len, flags);
if (res <= -4) {
fprintf(stderr, "Error running sam_prob_realn : %s\n",
strerror(errno));
goto fail;
}
}
if (sam_write1(out, hdr, rec) < 0) {
fprintf(stderr, "Error writing to %s\n", out_name);
goto fail;
}
}
res = hts_close(in);
in = NULL;
if (res < 0) {
fprintf(stderr, "Error closing %s\n", in_name);
goto fail;
}
res = hts_close(out);
out = NULL;
if (res < 0) {
fprintf(stderr, "Error closing %s\n", out_name);
goto fail;
}
bam_hdr_destroy(hdr);
bam_destroy1(rec);
free(ref_seq);
fai_destroy(fai);
return EXIT_SUCCESS;
fail:
if (hdr) bam_hdr_destroy(hdr);
if (rec) bam_destroy1(rec);
if (in) hts_close(in);
if (out) hts_close(out);
free(ref_seq);
fai_destroy(fai);
return EXIT_FAILURE;
}
/*!
@abstract Merge multiple sorted BAM.
@param is_by_qname whether to sort by query name
@param out output BAM file name
@param mode sam_open() mode to be used to create the final output file
(overrides level settings from UNCOMP and LEVEL1 flags)
@param headers name of SAM file from which to copy '@' header lines,
or NULL to copy them from the first file to be merged
@param n number of files to be merged
@param fn names of files to be merged
@param flag flags that control how the merge is undertaken
@param reg region to merge
@param n_threads number of threads to use (passed to htslib)
@discussion Padding information may NOT correctly maintained. This
function is NOT thread safe.
*/
int bam_merge_core2(int by_qname, const char *out, const char *mode, const char *headers, int n, char * const *fn, int flag, const char *reg, int n_threads)
{
samFile *fpout, **fp;
heap1_t *heap;
bam_hdr_t *hout = NULL;
int i, j, *RG_len = NULL;
uint64_t idx = 0;
char **RG = NULL;
hts_itr_t **iter = NULL;
bam_hdr_t **hdr = NULL;
trans_tbl_t *translation_tbl = NULL;
// Is there a specified pre-prepared header to use for output?
if (headers) {
samFile* fpheaders = sam_open(headers, "r");
if (fpheaders == NULL) {
const char *message = strerror(errno);
fprintf(pysamerr, "[bam_merge_core] cannot open '%s': %s\n", headers, message);
return -1;
}
hout = sam_hdr_read(fpheaders);
sam_close(fpheaders);
}
g_is_by_qname = by_qname;
fp = (samFile**)calloc(n, sizeof(samFile*));
heap = (heap1_t*)calloc(n, sizeof(heap1_t));
iter = (hts_itr_t**)calloc(n, sizeof(hts_itr_t*));
hdr = (bam_hdr_t**)calloc(n, sizeof(bam_hdr_t*));
translation_tbl = (trans_tbl_t*)calloc(n, sizeof(trans_tbl_t));
// prepare RG tag from file names
if (flag & MERGE_RG) {
RG = (char**)calloc(n, sizeof(char*));
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] = (char*)calloc(l + 1, 1);
RG_len[i] = l;
strncpy(RG[i], s + j, l);
}
}
// open and read the header from each file
for (i = 0; i < n; ++i) {
bam_hdr_t *hin;
fp[i] = sam_open(fn[i], "r");
if (fp[i] == NULL) {
int j;
fprintf(pysamerr, "[bam_merge_core] fail to open file %s\n", fn[i]);
for (j = 0; j < i; ++j) sam_close(fp[j]);
free(fp); free(heap);
// FIXME: possible memory leak
return -1;
}
hin = sam_hdr_read(fp[i]);
if (hout)
trans_tbl_init(hout, hin, translation_tbl+i, flag & MERGE_COMBINE_RG, flag & MERGE_COMBINE_PG);
else {
// As yet, no headers to merge into...
hout = bam_hdr_dup(hin);
// ...so no need to translate header into itself
trans_tbl_init(hout, hin, translation_tbl+i, true, true);
}
// TODO sam_itr_next() doesn't yet work for SAM files,
// so for those keep the headers around for use with sam_read1()
if (hts_get_format(fp[i])->format == sam) hdr[i] = hin;
else { bam_hdr_destroy(hin); hdr[i] = NULL; }
if ((translation_tbl+i)->lost_coord_sort && !by_qname) {
fprintf(pysamerr, "[bam_merge_core] Order of targets in file %s caused coordinate sort to be lost\n", fn[i]);
}
}
// Transform the header into standard form
pretty_header(&hout->text,hout->l_text);
// If we're only merging a specified region move our iters to start at that point
if (reg) {
int* rtrans = rtrans_build(n, hout->n_targets, translation_tbl);
int tid, beg, end;
const char *name_lim = hts_parse_reg(reg, &beg, &end);
char *name = malloc(name_lim - reg + 1);
memcpy(name, reg, name_lim - reg);
name[name_lim - reg] = '\0';
tid = bam_name2id(hout, name);
free(name);
if (tid < 0) {
fprintf(pysamerr, "[%s] Malformated region string or undefined reference name\n", __func__);
return -1;
}
for (i = 0; i < n; ++i) {
hts_idx_t *idx = sam_index_load(fp[i], fn[i]);
// (rtrans[i*n+tid]) Look up what hout tid translates to in input tid space
int mapped_tid = rtrans[i*hout->n_targets+tid];
if (mapped_tid != INT32_MIN) {
iter[i] = sam_itr_queryi(idx, mapped_tid, beg, end);
} else {
iter[i] = sam_itr_queryi(idx, HTS_IDX_NONE, 0, 0);
}
hts_idx_destroy(idx);
if (iter[i] == NULL) break;
}
free(rtrans);
} else {
for (i = 0; i < n; ++i) {
if (hdr[i] == NULL) {
iter[i] = sam_itr_queryi(NULL, HTS_IDX_REST, 0, 0);
if (iter[i] == NULL) break;
}
else iter[i] = NULL;
}
}
if (i < n) {
fprintf(pysamerr, "[%s] Memory allocation failed\n", __func__);
return -1;
}
// Load the first read from each file into the heap
for (i = 0; i < n; ++i) {
heap1_t *h = heap + i;
h->i = i;
h->b = bam_init1();
if ((iter[i]? sam_itr_next(fp[i], iter[i], h->b) : sam_read1(fp[i], hdr[i], h->b)) >= 0) {
bam_translate(h->b, translation_tbl + i);
h->pos = ((uint64_t)h->b->core.tid<<32) | (uint32_t)((int32_t)h->b->core.pos+1)<<1 | bam_is_rev(h->b);
h->idx = idx++;
}
else {
h->pos = HEAP_EMPTY;
bam_destroy1(h->b);
h->b = NULL;
}
}
// Open output file and write header
if ((fpout = sam_open(out, mode)) == 0) {
fprintf(pysamerr, "[%s] fail to create the output file.\n", __func__);
return -1;
}
sam_hdr_write(fpout, hout);
if (!(flag & MERGE_UNCOMP)) hts_set_threads(fpout, n_threads);
// Begin the actual merge
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]);
}
sam_write1(fpout, hout, b);
if ((j = (iter[heap->i]? sam_itr_next(fp[heap->i], iter[heap->i], b) : sam_read1(fp[heap->i], hdr[heap->i], b))) >= 0) {
bam_translate(b, translation_tbl + heap->i);
heap->pos = ((uint64_t)b->core.tid<<32) | (uint32_t)((int)b->core.pos+1)<<1 | bam_is_rev(b);
heap->idx = idx++;
} else if (j == -1) {
heap->pos = HEAP_EMPTY;
bam_destroy1(heap->b);
heap->b = NULL;
} else fprintf(pysamerr, "[bam_merge_core] '%s' is truncated. Continue anyway.\n", fn[heap->i]);
ks_heapadjust(heap, 0, n, heap);
}
// Clean up and close
if (flag & MERGE_RG) {
for (i = 0; i != n; ++i) free(RG[i]);
free(RG); free(RG_len);
}
for (i = 0; i < n; ++i) {
trans_tbl_destroy(translation_tbl + i);
hts_itr_destroy(iter[i]);
bam_hdr_destroy(hdr[i]);
sam_close(fp[i]);
}
bam_hdr_destroy(hout);
sam_close(fpout);
free(translation_tbl); free(fp); free(heap); free(iter); free(hdr);
return 0;
}
/******************************************************************************
*
* The main worker node function.
*
* int thread_id: the thread_id
* char *fastq1: FIFO from which bowtie2 can get read1
* char *fastq2: FIFO from which bowtie2 can get read2 (if it exists)
*
*******************************************************************************/
void herd_worker_node(int thread_id, char *fastq1, char *fastq2) {
int cmd_length = 1, max_qname = 0, status, strand;
char *cmd, *last_qname = calloc(1, sizeof(char));
MPI_Header *packed_header;
MPI_read *packed_read = calloc(1, sizeof(MPI_read));
bam_hdr_t *header;
bam1_t *read1 = bam_init1();
bam1_t *read2 = bam_init1();
samFile *fp;
#ifdef DEBUG
MPI_Status stat;
int current_p_size = 100;
htsFile *of;
bam_hdr_t *debug_header = bam_hdr_init();
bam1_t *debug_read = bam_init1();
global_header = bam_hdr_init();
void *p = calloc(100,1);
char *oname = NULL;
#else
int i = 0;
#endif
time_t t0, t1;
int swapped = 0;
assert(last_qname);
assert(packed_read);
//Which strand should we be aligning to?
if(config.directional) {
strand = (thread_id-1) % 2;
} else {
strand = (thread_id-1) % 4;
}
packed_read->size = 0;
packed_read->packed = NULL;
//construct the bowtie2 command
cmd_length += (int) strlen("bowtie2 -q --reorder") + 1;
cmd_length += (int) strlen(config.bowtie2_options) + 1;
cmd_length += (int) strlen("--norc -x") + 1;
cmd_length += (int) strlen(config.genome_dir) + strlen("bisulfite_genome/CT_conversion/BS_CT") + 1;
cmd_length += (int) 2*(strlen("-1 ") + strlen(fastq1)) + 3;
if(config.paired) cmd_length += (int) strlen(fastq2); //This is likely unneeded.
#ifdef DEBUG
oname = malloc(sizeof(char) *(1+strlen(config.odir)+strlen(config.basename)+strlen("_X.bam")));
assert(oname);
sprintf(oname, "%s%s_%i.bam", config.odir, config.basename, thread_id);
if(!config.quiet) fprintf(stderr, "Writing output to %s\n", oname);
of = sam_open(oname, "wb");
free(oname);
#endif
cmd = (char *) malloc(sizeof(char) * cmd_length);
assert(cmd);
if(strand == 0) { //OT Read#1 C->T, Read#2 G->A, Genome C->T only the + strand
if(config.paired) {
sprintf(cmd, "bowtie2 -q --reorder %s --norc -x %sbisulfite_genome/CT_conversion/BS_CT -1 %s -2 %s", config.bowtie2_options, config.genome_dir, fastq1, fastq2);
} else {
sprintf(cmd, "bowtie2 -q --reorder %s --norc -x %sbisulfite_genome/CT_conversion/BS_CT -U %s", config.bowtie2_options, config.genome_dir, fastq1);
}
} else if(strand == 1) { //OB Read#1 C->T, Read#2 G->A, Genome G->A only the - strand
if(config.paired) {
sprintf(cmd, "bowtie2 -q --reorder %s --nofw -x %sbisulfite_genome/GA_conversion/BS_GA -1 %s -2 %s", config.bowtie2_options, config.genome_dir, fastq1, fastq2);
} else {
sprintf(cmd, "bowtie2 -q --reorder %s --nofw -x %sbisulfite_genome/GA_conversion/BS_GA -U %s", config.bowtie2_options, config.genome_dir, fastq1);
}
} else if(strand == 2) { //CTOT Read#1 G->A, Read#2 C->T, Genome C->T, only the - strand
if(config.paired) {
sprintf(cmd, "bowtie2 -q --reorder %s --nofw -x %sbisulfite_genome/CT_conversion/BS_CT -1 %s -2 %s", config.bowtie2_options, config.genome_dir, fastq1, fastq2);
} else {
sprintf(cmd, "bowtie2 -q --reorder %s --nofw -x %sbisulfite_genome/CT_conversion/BS_CT -U %s", config.bowtie2_options, config.genome_dir, fastq1);
}
} else if(strand == 3) { //CTOB Read#1 G->A, Read#2 C->T, Genome G->A, only the + strand
if(config.paired) {
sprintf(cmd, "bowtie2 -q --reorder %s --norc -x %sbisulfite_genome/GA_conversion/BS_GA -1 %s -2 %s", config.bowtie2_options, config.genome_dir, fastq1, fastq2);
} else {
sprintf(cmd, "bowtie2 -q --reorder %s --norc -x %sbisulfite_genome/GA_conversion/BS_GA -U %s", config.bowtie2_options, config.genome_dir, fastq1);
}
} else {
fprintf(stderr, "Oh shit, got strand %i!\n", strand);
return;
}
//Start the process
if(!config.quiet) fprintf(stderr, "Node %i executing: %s\n", thread_id, cmd); fflush(stderr);
fp = sam_popen(cmd);
header = sam_hdr_read(fp);
#ifdef DEBUG
sam_hdr_write(of, header);
#endif
#ifndef DEBUG
packed_header = pack_header(header);
if(thread_id == 1) {
//Send the header
MPI_Send((void *) &(packed_header->size), 1, MPI_INT, 0, 1, MPI_COMM_WORLD);
status = MPI_Send((void *) packed_header->packed, packed_header->size, MPI_BYTE, 0, 2, MPI_COMM_WORLD);
if(status != MPI_SUCCESS) {
fprintf(stderr, "MPI_Send returned %i\n", status);
fflush(stderr);
}
}
#else
packed_header = pack_header(header);
void *tmp_pointer = malloc(packed_header->size);
assert(tmp_pointer);
MPI_Request request;
MPI_Isend((void *) packed_header->packed, packed_header->size, MPI_BYTE, 0, 2, MPI_COMM_WORLD, &request);
status = MPI_Recv(tmp_pointer, packed_header->size, MPI_BYTE, 0, 2, MPI_COMM_WORLD, &stat);
if(status != MPI_SUCCESS) fprintf(stderr, "We seem to have not been able to send the message to ourselves!\n");
MPI_Wait(&request, &stat);
unpack_header(debug_header, tmp_pointer);
global_header = debug_header;
free(tmp_pointer);
#endif
t0 = time(NULL);
if(!config.quiet) fprintf(stderr, "Node %i began sending reads @%s", thread_id, ctime(&t0)); fflush(stderr);
while(sam_read1(fp, header, read1) >= 0) {
#ifdef DEBUG
sam_write1(of, global_header, read1);
#endif
if(strcmp(bam_get_qname(read1), last_qname) == 0) { //Multimapper
if(config.paired) {
sam_read1(fp, header, read2);
#ifdef DEBUG
sam_write1(of, global_header, read2);
#endif
}
continue;
} else {
if(read1->core.l_qname > max_qname) {
max_qname = read1->core.l_qname + 10;
last_qname = realloc(last_qname, sizeof(char) * max_qname);
assert(last_qname);
}
strcpy(last_qname, bam_get_qname(read1));
}
//Are paired-end reads in the wrong order?
swapped = 0;
if(config.paired) {
if(read1->core.flag & BAM_FREAD2) {
swapped = 1;
sam_read1(fp, header, read2);
packed_read = pack_read(read2, packed_read);
#ifndef DEBUG
MPI_Send((void *) packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD);
#else
sam_write1(of, global_header, read2);
if(packed_read->size > current_p_size) {
p = realloc(p, packed_read->size);
assert(p);
}
MPI_Isend((void *) packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &request);
status = MPI_Recv(p, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &stat);
MPI_Wait(&request, &stat);
debug_read = unpack_read(debug_read, p);
#endif
}
}
//Send the read
packed_read = pack_read(read1, packed_read);
#ifndef DEBUG
MPI_Send((void *) packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD);
#else
if(packed_read->size > current_p_size) {
p = realloc(p, packed_read->size);
assert(p);
}
MPI_Isend(packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &request);
status = MPI_Recv(p, packed_header->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &stat);
MPI_Wait(&request, &stat);
#endif
//Deal with paired-end reads
if(config.paired && !swapped) {
sam_read1(fp, header, read2);
packed_read = pack_read(read2, packed_read);
#ifndef DEBUG
MPI_Send((void *) packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD);
#else
sam_write1(of, global_header, read2);
if(packed_read->size > current_p_size) {
p = realloc(p, packed_read->size);
assert(p);
}
MPI_Isend((void *) packed_read->packed, packed_read->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &request);
status = MPI_Recv(p, packed_header->size, MPI_BYTE, 0, 5, MPI_COMM_WORLD, &stat);
MPI_Wait(&request, &stat);
debug_read = unpack_read(debug_read, p);
#endif
}
#ifndef DEBUG
i++;
#endif
}
t1 = time(NULL);
if(!config.quiet) fprintf(stderr, "Node %i finished sending reads @%s\t(%f sec elapsed)\n", thread_id, ctime(&t1), difftime(t1, t0)); fflush(stderr);
//Notify the master node
packed_read->size = 0;
#ifndef DEBUG
void *A = malloc(1);
assert(A);
MPI_Send(A, 1, MPI_BYTE, 0, 5, MPI_COMM_WORLD);
free(A);
#endif
//Close things up
bam_hdr_destroy(header);
bam_destroy1(read1);
bam_destroy1(read2);
free(cmd);
if(packed_read->packed != NULL) free(packed_read->packed);
free(packed_read);
if(packed_header->packed != NULL) free(packed_header->packed);
free(packed_header);
free(last_qname);
sam_pclose(fp);
//Remove the FIFO(s)
unlink(fastq1);
if(config.paired) unlink(fastq2);
#ifdef DEBUG
sam_close(of);
bam_hdr_destroy(debug_header);
bam_destroy1(debug_read);
free(p);
#endif
if(!config.quiet) fprintf(stderr, "Exiting worker node %i\n", thread_id); fflush(stderr);
};