/* bam_split.c -- split subcommand. Copyright (C) 2013, 2014 Genome Research Ltd. Author: Martin Pollard Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions: The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software. THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE. */ #include #include #include #include #include #include #include #include #include #include KHASH_MAP_INIT_STR(c2i, int) struct parsed_opts { char* merged_input_name; char* unaccounted_header_name; char* unaccounted_name; char* output_format_string; bool verbose; }; typedef struct parsed_opts parsed_opts_t; struct state { samFile* merged_input_file; bam_hdr_t* merged_input_header; samFile* unaccounted_file; bam_hdr_t* unaccounted_header; size_t output_count; char** rg_id; samFile** rg_output_file; bam_hdr_t** rg_output_header; kh_c2i_t* rg_hash; }; typedef struct state state_t; static void cleanup_state(state_t* status); static void cleanup_opts(parsed_opts_t* opts); static void usage(FILE *write_to) { fprintf(write_to, "Usage: samtools split [-u [:]]\n" " [-f ] [-v] \n" "Options:\n" " -f STRING output filename format string [\"%%*_%%#.bam\"]\n" " -u FILE1 put reads with no RG tag or an unrecognised RG tag in FILE1\n" " -u FILE1:FILE2 ...and override the header with FILE2\n" " -v verbose output\n" "\n" "Format string expansions:\n" " %%%% %%\n" " %%* basename\n" " %%# @RG index\n" " %%! @RG ID\n" ); } // Takes the command line options and turns them into something we can understand static parsed_opts_t* parse_args(int argc, char** argv) { if (argc == 1) { usage(stdout); return NULL; } const char* optstring = "vf:u:"; char* delim; parsed_opts_t* retval = calloc(sizeof(parsed_opts_t), 1); if (! retval ) { perror("cannot allocate option parsing memory"); return NULL; } int opt; while ((opt = getopt(argc, argv, optstring)) != -1) { switch (opt) { case 'f': retval->output_format_string = strdup(optarg); if (! retval->output_format_string ) { perror("cannot allocate output format string memory"); return NULL; } break; case 'v': retval->verbose = true; break; case 'u': retval->unaccounted_name = strdup(optarg); if (! retval->unaccounted_name ) { perror("cannot allocate string memory"); return NULL; } if ((delim = strchr(retval->unaccounted_name, ':')) != NULL) { *delim = '\0'; retval->unaccounted_header_name = strdup(delim+1); if (! retval->unaccounted_header_name ) { perror("cannot allocate string memory"); return NULL; } } break; default: usage(stdout); free(retval); return NULL; } } if (retval->output_format_string == NULL) retval->output_format_string = strdup("%*_%#.bam"); argc -= optind; argv += optind; if (argc != 1) { fprintf(stderr, "Invalid number of arguments: %d\n", argc); usage(stderr); free(retval); return NULL; } retval->merged_input_name = strdup(argv[0]); if (! retval->merged_input_name ) { perror("cannot allocate string memory"); return NULL; } return retval; } // Expands a output filename format string static char* expand_format_string(const char* format_string, const char* basename, const char* rg_id, const int rg_idx) { kstring_t str = { 0, 0, NULL }; const char* pointer = format_string; const char* next; while ((next = strchr(pointer, '%')) != NULL) { kputsn(pointer, next-pointer, &str); ++next; switch (*next) { case '%': kputc('%', &str); break; case '*': kputs(basename, &str); break; case '#': kputl(rg_idx, &str); break; case '!': kputs(rg_id, &str); break; case '\0': // Error is: fprintf(stderr, "bad format string, trailing %%\n"); free(str.s); return NULL; default: // Error is: fprintf(stderr, "bad format string, unknown format specifier\n"); free(str.s); return NULL; } pointer = next + 1; } kputs(pointer, &str); return ks_release(&str); } // Parse the header, count the number of RG tags and return a list of their names static bool count_RG(bam_hdr_t* hdr, size_t* count, char*** output_name) { if (hdr->l_text < 3 ) { *count = 0; *output_name = NULL; return true; } kstring_t input = { 0, 0, NULL }; kputsn(hdr->text, hdr->l_text, &input); ////////////////////////////////////////// // First stage count number of @RG tags // ////////////////////////////////////////// char* pointer = ks_str(&input); size_t n_rg = 0; // Guard against rare case where @RG is first header line // This shouldn't happen but could where @HD is omitted if (pointer[0] == '@' && pointer[1] == 'R' && pointer[2] == 'G' ) { ++n_rg; pointer += 3; } char* line; while ((line = strstr(pointer, "\n@RG")) != NULL) { ++n_rg; pointer = line + 1; } ////////////////////////////////// // Second stage locate @RG ID's // ////////////////////////////////// char** names = (char**)calloc(sizeof(char*), n_rg); size_t next = 0; regex_t rg_finder; if (regcomp(&rg_finder, "^@RG.*\tID:([!-)+-<>-~][ !-~]*)(\t.*$|$)", REG_EXTENDED|REG_NEWLINE) != 0) { free(input.s); free(names); return false; } regmatch_t* matches = (regmatch_t*)calloc(sizeof(regmatch_t),2); int error; char* begin = ks_str(&input); while ((error = regexec(&rg_finder, begin, 2, matches, 0)) == 0) { kstring_t str = { 0, 0, NULL }; kputsn(begin+matches[1].rm_so, matches[1].rm_eo-matches[1].rm_so, &str); names[next++] = ks_release(&str); begin += matches[0].rm_eo; } if (error != REG_NOMATCH) { // cleanup regfree(&rg_finder); free(matches); free(names); free(input.s); return false; } free(matches); // return results *count = n_rg; *output_name = names; regfree(&rg_finder); free(input.s); return true; } // Filters a header of @RG lines where ID != id_keep // TODO: strip @PG's descended from other RGs and their descendants static bool filter_header_rg(bam_hdr_t* hdr, const char* id_keep) { kstring_t str = {0, 0, NULL}; regex_t rg_finder; if (regcomp(&rg_finder, "^@RG.*\tID:([!-)+-<>-~][ !-~]*)(\t.*$|$)", REG_EXTENDED|REG_NEWLINE) != 0) { return false; } // regex vars char* header = hdr->text; regmatch_t* matches = (regmatch_t*)calloc(sizeof(regmatch_t),2); kstring_t found_id = { 0, 0, NULL }; int error; while ((error = regexec(&rg_finder, header, 2, matches, 0)) == 0) { kputsn(header, matches[0].rm_so, &str); // copy header up until the found RG line found_id.l = 0; kputsn(header+matches[1].rm_so, matches[1].rm_eo-matches[1].rm_so, &found_id); // extract ID // if it matches keep keep it, else we can just ignore it if (strcmp(ks_str(&found_id), id_keep) == 0) { kputsn(header+matches[0].rm_so, (matches[0].rm_eo+1)-matches[0].rm_so, &str); } // move pointer forward header += matches[0].rm_eo+1; } // cleanup free(found_id.s); free(matches); regfree(&rg_finder); // Did we leave loop because of an error? if (error != REG_NOMATCH) { return false; } // Write remainder of string kputs(header, &str); // Modify header hdr->l_text = ks_len(&str); free(hdr->text); hdr->text = ks_release(&str); return true; } // Set the initial state static state_t* init(parsed_opts_t* opts) { state_t* retval = calloc(sizeof(state_t), 1); if (!retval) { fprintf(stderr, "Out of memory"); return NULL; } retval->merged_input_file = sam_open(opts->merged_input_name, "rb"); if (!retval->merged_input_file) { fprintf(stderr, "Could not open input file (%s)\n", opts->merged_input_name); free(retval); return NULL; } retval->merged_input_header = sam_hdr_read(retval->merged_input_file); if (opts->unaccounted_name) { if (opts->unaccounted_header_name) { samFile* hdr_load = sam_open(opts->unaccounted_header_name, "r"); if (!hdr_load) { fprintf(stderr, "Could not open unaccounted header file (%s)\n", opts->unaccounted_header_name); cleanup_state(retval); return NULL; } retval->unaccounted_header = sam_hdr_read(hdr_load); sam_close(hdr_load); } else { retval->unaccounted_header = bam_hdr_dup(retval->merged_input_header); } retval->unaccounted_file = sam_open(opts->unaccounted_name, "wb"); if (retval->unaccounted_file == NULL) { fprintf(stderr, "Could not open unaccounted output file: %s\n", opts->unaccounted_name); cleanup_state(retval); return NULL; } } // Open output files for RGs if (!count_RG(retval->merged_input_header, &retval->output_count, &retval->rg_id)) return NULL; if (opts->verbose) fprintf(stderr, "@RG's found %zu\n",retval->output_count); retval->rg_output_file = (samFile**)calloc(retval->output_count, sizeof(samFile*)); retval->rg_output_header = (bam_hdr_t**)calloc(retval->output_count, sizeof(bam_hdr_t*)); retval->rg_hash = kh_init_c2i(); if (!retval->rg_output_file || !retval->rg_output_header) { fprintf(stderr, "Could not allocate memory for output file array. Out of memory?"); cleanup_state(retval); return NULL; } char* dirsep = strrchr(opts->merged_input_name, '/'); char* input_base_name = strdup(dirsep? dirsep+1 : opts->merged_input_name); if (!input_base_name) { fprintf(stderr, "Out of memory\n"); cleanup_state(retval); return NULL; } char* extension = strrchr(input_base_name, '.'); if (extension) *extension = '\0'; size_t i; for (i = 0; i < retval->output_count; i++) { char* output_filename = NULL; if ( ( output_filename = expand_format_string(opts->output_format_string, input_base_name, retval->rg_id[i], i) ) == NULL) { fprintf(stderr, "Error expanding output filename format string.\r\n"); cleanup_state(retval); free(input_base_name); return NULL; } retval->rg_output_file[i] = sam_open(output_filename, "wb"); if (retval->rg_output_file[i] == NULL) { fprintf(stderr, "Could not open output file: %s\r\n", output_filename); cleanup_state(retval); free(input_base_name); return NULL; } // Record index in hash int ret; khiter_t iter = kh_put_c2i(retval->rg_hash, retval->rg_id[i], &ret); kh_val(retval->rg_hash,iter) = i; // Set and edit header retval->rg_output_header[i] = bam_hdr_dup(retval->merged_input_header); if ( !filter_header_rg(retval->rg_output_header[i], retval->rg_id[i]) ) { fprintf(stderr, "Could not rewrite header for file: %s\r\n", output_filename); cleanup_state(retval); free(output_filename); free(input_base_name); return NULL; } free(output_filename); } free(input_base_name); return retval; } static bool split(state_t* state) { if (state->unaccounted_file && sam_hdr_write(state->unaccounted_file, state->unaccounted_header) != 0) { fprintf(stderr, "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(stderr, "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(stderr, "Read \"%s\" with unaccounted for tag \"%s\".\n", bam_get_qname(file_read), bam_aux2Z(tag)); } else { fprintf(stderr, "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; } static void cleanup_state(state_t* status) { if (!status) return; if (status->unaccounted_header) bam_hdr_destroy(status->unaccounted_header); if (status->unaccounted_file) sam_close(status->unaccounted_file); sam_close(status->merged_input_file); size_t i; for (i = 0; i < status->output_count; i++) { bam_hdr_destroy(status->rg_output_header[i]); sam_close(status->rg_output_file[i]); free(status->rg_id[i]); } bam_hdr_destroy(status->merged_input_header); free(status->rg_output_header); free(status->rg_output_file); kh_destroy_c2i(status->rg_hash); free(status->rg_id); free(status); } static void cleanup_opts(parsed_opts_t* opts) { if (!opts) return; free(opts->merged_input_name); free(opts->unaccounted_header_name); free(opts->unaccounted_name); free(opts->output_format_string); free(opts); } int main_split(int argc, char** argv) { int ret = 1; parsed_opts_t* opts = parse_args(argc, argv); if (!opts ) goto cleanup_opts; state_t* status = init(opts); if (!status) goto cleanup_opts; if (split(status)) ret = 0; cleanup_state(status); cleanup_opts: cleanup_opts(opts); return ret; }