#define __STDC_LIMIT_MACROS #include #include #include #include #include #include #include "CommandLines.h" #include "ketopt.h" #define DEFAULT_OUTPUT "hifiasm.asm" hifiasm_opt_t asm_opt; static ko_longopt_t long_options[] = { { "version", ko_no_argument, 300 }, { "dbg-gfa", ko_no_argument, 301 }, { "write-paf", ko_no_argument, 302 }, { "write-ec", ko_no_argument, 303 }, { "skip-triobin", ko_no_argument, 304 }, { "max-od-ec", ko_no_argument, 305 }, { "max-od-final", ko_no_argument, 306 }, { "ex-list", ko_required_argument, 307 }, { "ex-iter", ko_required_argument, 308 }, { "hom-cov", ko_required_argument, 309 }, { "pri-range", ko_required_argument, 310 }, { "lowQ", ko_required_argument, 312 }, { "min-hist-cnt", ko_required_argument, 313 }, { "h1", ko_required_argument, 314 }, { "h2", ko_required_argument, 315 }, { "enzyme", ko_required_argument, 316 }, { "b-cov", ko_required_argument, 317 }, { "h-cov", ko_required_argument, 318 }, { "m-rate", ko_required_argument, 319 }, { "primary", ko_no_argument, 320 }, { "t-occ", ko_required_argument, 321 }, { "seed", ko_required_argument, 322 }, { "n-perturb", ko_required_argument, 323 }, { "f-perturb", ko_required_argument, 324 }, { "n-hap", ko_required_argument, 325 }, { "n-weight", ko_required_argument, 326 }, { "l-msjoin", ko_required_argument, 327 }, { "purge-max", ko_required_argument, 328 }, { "fast", ko_no_argument, 329 }, { "dp-er", ko_required_argument, 330}, { "max-kocc", ko_required_argument, 331}, { "hg-size", ko_required_argument, 332}, { "ul", ko_required_argument, 333}, { "unskew", ko_no_argument, 334}, { "kpt-rate", ko_required_argument, 335}, { "ul-rate", ko_required_argument, 336}, { "dbg-het-cnt", ko_no_argument, 337}, { "ul-tip", ko_required_argument, 338}, { "low-het", ko_no_argument, 339}, { "s-base", ko_required_argument, 340}, { "bin-only", ko_no_argument, 341}, { "ul-round", ko_required_argument, 342}, { "prt-raw", ko_no_argument, 343}, { "integer-correct", ko_required_argument, 344}, { "dbg-ovec", ko_no_argument, 345}, { "path-max", ko_required_argument, 346}, { "path-min", ko_required_argument, 347}, { "trio-dual", ko_no_argument, 348}, { "ul-cut", ko_required_argument, 349}, { "dual-scaf", ko_no_argument, 350}, { "scaf-gap", ko_required_argument, 351}, // { "path-round", ko_required_argument, 348}, { 0, 0, 0 } }; double Get_T(void) { struct timeval t; gettimeofday(&t, NULL); return t.tv_sec+t.tv_usec/1000000.0; } void Print_H(hifiasm_opt_t* asm_opt) { fprintf(stderr, "Usage: hifiasm [options] <...>\n"); fprintf(stderr, "Options:\n"); fprintf(stderr, " Input/Output:\n"); fprintf(stderr, " -o STR prefix of output files [%s]\n", asm_opt->output_file_name); fprintf(stderr, " -t INT number of threads [%d]\n", asm_opt->thread_num); fprintf(stderr, " -h show help information\n"); fprintf(stderr, " --version show version number\n"); fprintf(stderr, " Overlap/Error correction:\n"); fprintf(stderr, " -k INT k-mer length (must be <64) [%d]\n", asm_opt->k_mer_length); fprintf(stderr, " -w INT minimizer window size [%d]\n", asm_opt->mz_win); fprintf(stderr, " -f INT number of bits for bloom filter; 0 to disable [%d]\n", asm_opt->bf_shift); fprintf(stderr, " -D FLOAT drop k-mers occurring >FLOAT*coverage times [%.1f]\n", asm_opt->high_factor); fprintf(stderr, " -N INT consider up to max(-D*coverage,-N) overlaps for each oriented read [%d]\n", asm_opt->max_n_chain); fprintf(stderr, " -r INT round of correction [%d]\n", asm_opt->number_of_round); fprintf(stderr, " -z INT length of adapters that should be removed [%d]\n", asm_opt->adapterLen); fprintf(stderr, " --max-kocc INT\n"); fprintf(stderr, " employ k-mers occurring max_kmer_cnt); fprintf(stderr, " --hg-size INT(k, m or g)\n"); fprintf(stderr, " estimated haploid genome size used for inferring read coverage [auto]\n"); fprintf(stderr, " Assembly:\n"); fprintf(stderr, " -a INT round of assembly cleaning [%d]\n", asm_opt->clean_round); fprintf(stderr, " -m INT pop bubbles of large_pop_bubble_size); fprintf(stderr, " -p INT pop bubbles of small_pop_bubble_size); fprintf(stderr, " -n INT remove tip unitigs composed of <=INT reads [%d]\n", asm_opt->max_short_tip); fprintf(stderr, " -x FLOAT max overlap drop ratio [%.2g]\n", asm_opt->max_drop_rate); fprintf(stderr, " -y FLOAT min overlap drop ratio [%.2g]\n", asm_opt->min_drop_rate); fprintf(stderr, " -i ignore saved read correction and overlaps\n"); fprintf(stderr, " -u post-join step for contigs which may improve N50; 0 to disable; 1 to enable\n"); fprintf(stderr, " [%u] and [%u] in default for the UL+HiFi assembly and the HiFi assembly, respectively\n", asm_opt->ul_pst_join, asm_opt->hifi_pst_join); fprintf(stderr, " --hom-cov INT\n"); fprintf(stderr, " homozygous read coverage [auto]\n"); fprintf(stderr, " --lowQ INT\n"); fprintf(stderr, " output contig regions with >=INT%% inconsistency in BED format; 0 to disable [%d]\n", asm_opt->bed_inconsist_rate); fprintf(stderr, " --b-cov INT\n"); fprintf(stderr, " break contigs at positions with b_low_cov); fprintf(stderr, " --h-cov INT\n"); fprintf(stderr, " break contigs at positions with >INT-fold coverage; work with '--m-rate'; -1 to disable [%d]\n", asm_opt->b_high_cov); fprintf(stderr, " --m-rate FLOAT\n"); fprintf(stderr, " break contigs at positions with <=FLOAT*coverage exact overlaps;\n"); fprintf(stderr, " only work with '--b-cov' or '--h-cov'[%.2f]\n", asm_opt->m_rate); fprintf(stderr, " --primary output a primary assembly and an alternate assembly\n"); // fprintf(stderr, " --pri-range INT1[,INT2]\n"); // fprintf(stderr, " keep contigs with coverage in this range in p_ctg.gfa; -1 to disable [auto,inf]\n"); fprintf(stderr, " Trio-partition:\n"); fprintf(stderr, " -1 FILE hap1/paternal k-mer dump generated by \"yak count\" []\n"); fprintf(stderr, " -2 FILE hap2/maternal k-mer dump generated by \"yak count\" []\n"); fprintf(stderr, " -3 FILE list of hap1/paternal read names []\n"); fprintf(stderr, " -4 FILE list of hap2/maternal read names []\n"); fprintf(stderr, " -c INT lower bound of the binned k-mer's frequency [%d]\n", asm_opt->min_cnt); fprintf(stderr, " -d INT upper bound of the binned k-mer's frequency [%d]\n", asm_opt->mid_cnt); fprintf(stderr, " --t-occ INT\n"); fprintf(stderr, " forcedly remove unitigs with >INT unexpected haplotype-specific reads;\n"); fprintf(stderr, " ignore graph topology; [%d]\n", asm_opt->trio_flag_occ_thres); fprintf(stderr, " --trio-dual utilize homology information to correct trio phasing errors\n"); fprintf(stderr, " Purge-dups:\n"); fprintf(stderr, " -l INT purge level. 0: no purging; 1: light; 2/3: aggressive [0 for trio; 3 for unzip]\n"); fprintf(stderr, " -s FLOAT similarity threshold for duplicate haplotigs in read-level [%g for -l1/-l2, %g for -l3]\n", asm_opt->purge_simi_rate_l2, asm_opt->purge_simi_rate_l3); fprintf(stderr, " -O INT min number of overlapped reads for duplicate haplotigs [%d]\n", asm_opt->purge_overlap_len); fprintf(stderr, " --purge-max INT\n"); fprintf(stderr, " coverage upper bound of Purge-dups [auto]\n"); fprintf(stderr, " --n-hap INT\n"); fprintf(stderr, " number of haplotypes [%d]\n", asm_opt->polyploidy); // fprintf(stderr, " Hi-C-partition [experimental, not stable]:\n"); fprintf(stderr, " Hi-C-partition:\n"); fprintf(stderr, " --h1 FILEs file names of Hi-C R1 [r1_1.fq,r1_2.fq,...]\n"); fprintf(stderr, " --h2 FILEs file names of Hi-C R2 [r2_1.fq,r2_2.fq,...]\n"); fprintf(stderr, " --seed INT RNG seed [%lu]\n", asm_opt->seed); fprintf(stderr, " --s-base FLOAT\n"); fprintf(stderr, " similarity threshold for homology detection in base-level;\n"); fprintf(stderr, " -1 to disable [%.3g]; -s for read-level (see )\n", asm_opt->trans_base_rate_sec); fprintf(stderr, " --n-weight INT\n"); fprintf(stderr, " rounds of reweighting Hi-C links [%d]\n", asm_opt->n_weight); fprintf(stderr, " --n-perturb INT\n"); fprintf(stderr, " rounds of perturbation [%d]\n", asm_opt->n_perturb); fprintf(stderr, " --f-perturb FLOAT\n"); fprintf(stderr, " fraction to flip for perturbation [%.3g]\n", asm_opt->f_perturb); fprintf(stderr, " --l-msjoin INT\n"); fprintf(stderr, " detect misjoined unitigs of >=INT in size; 0 to disable [%lu]\n", asm_opt->misjoin_len); fprintf(stderr, " Ultra-Long-integration:\n"); fprintf(stderr, " --ul FILEs file names of Ultra-Long reads [r1.fq,r2.fq,...]\n"); fprintf(stderr, " --ul-rate FLOAT\n"); fprintf(stderr, " error rate of Ultra-Long reads [%.3g]\n", asm_opt->ul_error_rate); fprintf(stderr, " --ul-tip INT\n"); fprintf(stderr, " remove tip unitigs composed of <=INT reads for the UL assembly [%d]\n", asm_opt->max_short_ul_tip); fprintf(stderr, " --path-max FLOAT\n"); fprintf(stderr, " max path drop ratio [%.2g]; higher number may make the assembly cleaner\n", asm_opt->max_path_drop_rate); fprintf(stderr, " but may lead to more misassemblies\n"); fprintf(stderr, " --path-min FLOAT\n"); fprintf(stderr, " min path drop ratio [%.2g]; higher number may make the assembly cleaner\n", asm_opt->min_path_drop_rate); fprintf(stderr, " but may lead to more misassemblies\n"); fprintf(stderr, " --ul-cut INT\n"); fprintf(stderr, " filter out ul_min_base); // fprintf(stderr, " --low-het enable it for genomes with very low het heterozygosity rate (<0.0001%%)\n"); fprintf(stderr, " Dual-Scaffolding:\n"); fprintf(stderr, " --dual-scaf output scaffolding\n"); fprintf(stderr, " --scaf-gap INT\n"); fprintf(stderr, " max gap size for scaffolding [%ld]\n", asm_opt->self_scaf_gap_max); fprintf(stderr, "Example: ./hifiasm -o NA12878.asm -t 32 NA12878.fq.gz\n"); fprintf(stderr, "See `https://hifiasm.readthedocs.io/en/latest/' or `man ./hifiasm.1' for complete documentation.\n"); } void init_opt(hifiasm_opt_t* asm_opt) { memset(asm_opt, 0, sizeof(hifiasm_opt_t)); ///asm_opt->flag = 0; asm_opt->flag = HA_F_PARTITION; asm_opt->coverage = -1; asm_opt->num_reads = 0; asm_opt->read_file_names = NULL; asm_opt->output_file_name = (char*)(DEFAULT_OUTPUT); asm_opt->required_read_name = NULL; asm_opt->hic_enzymes = NULL; asm_opt->hic_reads[0] = NULL; asm_opt->hic_reads[1] = NULL; asm_opt->fn_bin_poy = NULL; asm_opt->ar = NULL; asm_opt->thread_num = 1; asm_opt->k_mer_length = 51; asm_opt->hic_mer_length = 31; asm_opt->ul_mer_length = 19; asm_opt->trans_mer_length = 31; asm_opt->mz_win = 51; asm_opt->ul_mz_win = 19; asm_opt->trans_win = 31; asm_opt->mz_rewin = 1000; asm_opt->ul_mz_rewin = 360; asm_opt->mz_sample_dist = 500; asm_opt->bf_shift = 37; asm_opt->max_kmer_cnt = 2000; asm_opt->high_factor = 5.0; asm_opt->max_ov_diff_ec = 0.04; asm_opt->max_ov_diff_final = 0.03; asm_opt->hom_cov = 20; asm_opt->het_cov = -1024; asm_opt->max_n_chain = MIN_N_CHAIN; asm_opt->min_hist_kmer_cnt = 5; asm_opt->load_index_from_disk = 1; asm_opt->write_index_to_disk = 1; asm_opt->number_of_round = 3; asm_opt->adapterLen = 0; asm_opt->clean_round = 4; ///asm_opt->small_pop_bubble_size = 100000; asm_opt->small_pop_bubble_size = 0; asm_opt->large_pop_bubble_size = 10000000; asm_opt->min_drop_rate = 0.2; asm_opt->max_drop_rate = 0.8; asm_opt->max_hang_Len = 1000; asm_opt->max_hang_rate = 0.8; asm_opt->gap_fuzz = 1000; asm_opt->min_overlap_Len = 50; asm_opt->min_overlap_coverage = 0; asm_opt->max_short_tip = 3; asm_opt->max_short_ul_tip = 6; asm_opt->min_cnt = 2; asm_opt->mid_cnt = 5; asm_opt->purge_level_primary = 3; asm_opt->purge_level_trio = 0; asm_opt->purge_simi_rate_l2 = 0.75; asm_opt->purge_simi_rate_l3 = 0.55; asm_opt->trans_base_rate = 0.93; asm_opt->trans_base_rate_sec = 0.5; asm_opt->purge_overlap_len = 1; ///asm_opt->purge_overlap_len_hic = 50; asm_opt->recover_atg_cov_min = -1024; asm_opt->recover_atg_cov_max = INT_MAX; asm_opt->hom_global_coverage = -1; asm_opt->hom_global_coverage_set = 0; asm_opt->pur_global_coverage = -1; asm_opt->bed_inconsist_rate = 70; asm_opt->hic_inconsist_rate = 30; ///asm_opt->bub_mer_length = 3; asm_opt->bub_mer_length = 1000000; asm_opt->b_low_cov = 0; asm_opt->b_high_cov = -1; asm_opt->m_rate = 0.75; asm_opt->hap_occ = 1; asm_opt->polyploidy = 2; asm_opt->trio_flag_occ_thres = 60; asm_opt->seed = 11; asm_opt->n_perturb = 10000; asm_opt->f_perturb = 0.1; asm_opt->n_weight = 3; asm_opt->is_alt = 0; asm_opt->misjoin_len = 500000; asm_opt->scffold = 0; asm_opt->dp_min_len = 2000; asm_opt->dp_e = 0.0025; asm_opt->hg_size = -1; asm_opt->kpt_rate = -1; asm_opt->infor_cov = 3; asm_opt->s_hap_cov = 3; asm_opt->ul_error_rate = 0.2/**0.15**/; asm_opt->ul_error_rate_low = 0.1; asm_opt->ul_error_rate_hpc = 0.2; asm_opt->ul_ec_round = 3; asm_opt->is_dbg_het_cnt = 0; asm_opt->is_low_het_ul = 0; asm_opt->is_base_trans = 1; asm_opt->is_read_trans = 1; asm_opt->is_topo_trans = 1; asm_opt->is_bub_trans = 1; asm_opt->bin_only = 0; asm_opt->ul_clean_round = 1; asm_opt->prt_dbg_gfa = 0; asm_opt->integer_correct_round = 0; asm_opt->dbg_ovec_cal = 0; asm_opt->min_path_drop_rate = 0.2; asm_opt->max_path_drop_rate = 0.6; asm_opt->hifi_pst_join = 1; asm_opt->ul_pst_join = 1; asm_opt->trio_cov_het_ovlp = -1; asm_opt->ul_min_base = 0; asm_opt->self_scaf = 0; asm_opt->self_scaf_min = 250000; asm_opt->self_scaf_reliable_min = 5000000; asm_opt->self_scaf_gap_max = 3000000; } void destory_enzyme(enzyme* f) { int i; if(f != NULL) { for (i = 0; i < f->n; i++) { free(f->a[i]); } free(f->a); free(f->l); free(f); } } void destory_opt(hifiasm_opt_t* asm_opt) { if(asm_opt->read_file_names != NULL) free(asm_opt->read_file_names); if(asm_opt->hic_enzymes != NULL) destory_enzyme(asm_opt->hic_enzymes); if(asm_opt->hic_reads[0] != NULL) destory_enzyme(asm_opt->hic_reads[0]); if(asm_opt->hic_reads[1] != NULL) destory_enzyme(asm_opt->hic_reads[1]); if(asm_opt->ar != NULL) destory_enzyme(asm_opt->ar); } void ha_opt_reset_to_round(hifiasm_opt_t* asm_opt, int round) { asm_opt->num_bases = 0; asm_opt->num_corrected_bases = 0; asm_opt->num_recorrected_bases = 0; asm_opt->mem_buf = 0; asm_opt->roundID = round; } void ha_opt_update_cov(hifiasm_opt_t *opt, int hom_cov) { int max_n_chain = (int)(hom_cov * opt->high_factor + .499); opt->hom_cov = hom_cov; if (opt->max_n_chain < max_n_chain) opt->max_n_chain = max_n_chain; fprintf(stderr, "[M::%s] updated max_n_chain to %d\n", __func__, opt->max_n_chain); } void ha_opt_update_cov_min(hifiasm_opt_t *opt, int hom_cov, int min_chain) { int max_n_chain = (int)(hom_cov * opt->high_factor + .499); opt->hom_cov = hom_cov; opt->max_n_chain = max_n_chain; if(opt->max_n_chain < min_chain) opt->max_n_chain = min_chain; fprintf(stderr, "[M::%s] updated max_n_chain to %d\n", __func__, opt->max_n_chain); } static int check_file(char* name, const char* opt) { if(!name) { fprintf(stderr, "[ERROR] file does not exist (-%s)\n", opt); return 0; } FILE* is_exist = NULL; is_exist = fopen(name,"r"); if(!is_exist) { fprintf(stderr, "[ERROR] %s does not exist (-%s)\n", name, opt); return 0; } fclose(is_exist); return 1; } static int check_hic_reads(enzyme* f, const char* opt) { int i; for (i = 0; i < f->n; i++) { if(check_file(f->a[i], opt) == 0) return 0; } return 1; } int check_option(hifiasm_opt_t* asm_opt) { if(asm_opt->read_file_names == NULL || asm_opt->num_reads == 0) { fprintf(stderr, "[ERROR] missing input: please specify a read file\n"); return 0; } if(asm_opt->output_file_name == NULL) { fprintf(stderr, "[ERROR] missing output: please specify the output name (-o)\n"); return 0; } if(asm_opt->thread_num < 1) { fprintf(stderr, "[ERROR] the number of threads must be > 0 (-t)\n"); return 0; } if(asm_opt->number_of_round < 1) { fprintf(stderr, "[ERROR] the number of rounds for correction must be > 0 (-r)\n"); return 0; } if(asm_opt->clean_round < 1) { fprintf(stderr, "[ERROR] the number of rounds for assembly cleaning must be > 0 (-a)\n"); return 0; } if(asm_opt->adapterLen < 0) { fprintf(stderr, "[ERROR] the length of removed adapters must be >= 0 (-z)\n"); return 0; } if(asm_opt->k_mer_length >= 64) { fprintf(stderr, "[ERROR] the length of k_mer must be < 64 (-k)\n"); return 0; } if(asm_opt->max_drop_rate < 0 || asm_opt->max_drop_rate >= 1 ) { fprintf(stderr, "[ERROR] max overlap drop ratio must be [0.0, 1.0) (-x)\n"); return 0; } if(asm_opt->min_drop_rate < 0 || asm_opt->min_drop_rate >= 1) { fprintf(stderr, "[ERROR] min overlap drop ratio must be [0.0, 1.0) (-y)\n"); return 0; } if(asm_opt->max_drop_rate <= asm_opt->min_drop_rate) { fprintf(stderr, "[ERROR] min overlap drop ratio must be less than max overlap drop ratio (-x/-y)\n"); return 0; } if(asm_opt->small_pop_bubble_size < 0) { fprintf(stderr, "[ERROR] the size of popped small bubbles must be >= 0 (-p)\n"); return 0; } if(asm_opt->large_pop_bubble_size < 0) { fprintf(stderr, "[ERROR] the size of popped large bubbles must be >= 0 (-m)\n"); return 0; } if(asm_opt->max_hang_Len < 0) { fprintf(stderr, "[ERROR] max_hang_Len must be >= 0\n"); return 0; } if(asm_opt->max_hang_rate < 0) { fprintf(stderr, "[ERROR] max_hang_rate must be >= 0\n"); return 0; } if(asm_opt->gap_fuzz < 0) { fprintf(stderr, "[ERROR] gap_fuzz must be >= 0\n"); return 0; } if(asm_opt->min_overlap_Len < 0) { fprintf(stderr, "[ERROR] min_overlap_Len must be >= 0\n"); return 0; } if(asm_opt->min_overlap_coverage < 0) { fprintf(stderr, "[ERROR] min_overlap_coverage must be >= 0\n"); return 0; } if (asm_opt->max_ov_diff_ec < asm_opt->max_ov_diff_final) { fprintf(stderr, "[ERROR] max_ov_diff_ec shouldn't be smaller than max_ov_diff_final\n"); return 0; } if (asm_opt->max_ov_diff_ec < HA_MIN_OV_DIFF) { fprintf(stderr, "[ERROR] max_ov_diff_ec shouldn't be smaller than %g\n", HA_MIN_OV_DIFF); return 0; } if(asm_opt->max_short_tip < 0) { fprintf(stderr, "[ERROR] the length of removal tips must be >= 0 (-n)\n"); return 0; } if(asm_opt->purge_level_primary < 0 || asm_opt->purge_level_primary > 3) { fprintf(stderr, "[ERROR] the level of purge-dup should be [0, 3] (-l)\n"); return 0; } if(ha_opt_triobin(asm_opt) && ((asm_opt->purge_level_trio < 0 || asm_opt->purge_level_trio > 1))) { fprintf(stderr, "[ERROR] the level of purge-dup for trio should be [0, 1] (-l)\n"); return 0; } if(asm_opt->hom_global_coverage < 0 && asm_opt->hom_global_coverage != -1) { fprintf(stderr, "[ERROR] homozygous read coverage should be >= 0 (--hom-cov)\n"); return 0; } if(asm_opt->pur_global_coverage < 0 && asm_opt->pur_global_coverage != -1) { fprintf(stderr, "[ERROR] purge duplication coverage threshold should be >= 0 (--purge-max)\n"); return 0; } if(asm_opt->bed_inconsist_rate < 0 || asm_opt->bed_inconsist_rate > 100) { fprintf(stderr, "[ERROR] inconsistency rate should be [0, 100] (--lowQ)\n"); return 0; } if(asm_opt->fn_bin_yak[0] != NULL && check_file(asm_opt->fn_bin_yak[0], "YAK1") == 0) return 0; if(asm_opt->fn_bin_yak[1] != NULL && check_file(asm_opt->fn_bin_yak[1], "YAK2") == 0) return 0; if(asm_opt->fn_bin_list[0] != NULL && check_file(asm_opt->fn_bin_list[0], "LIST1") == 0) return 0; if(asm_opt->fn_bin_list[1] != NULL && check_file(asm_opt->fn_bin_list[1], "LIST2") == 0) return 0; if(asm_opt->required_read_name != NULL && check_file(asm_opt->required_read_name, "b") == 0) return 0; if(asm_opt->hic_reads[0] != NULL && check_hic_reads(asm_opt->hic_reads[0], "HIC1") == 0) return 0; if(asm_opt->hic_reads[1] != NULL && check_hic_reads(asm_opt->hic_reads[1], "HIC2") == 0) return 0; if(asm_opt->hic_reads[0] != NULL && asm_opt->hic_reads[1] == NULL) { fprintf(stderr, "[ERROR] lack r2 of HiC reads (--h2)\n"); return 0; } if(asm_opt->hic_reads[1] != NULL && asm_opt->hic_reads[0] == NULL) { fprintf(stderr, "[ERROR] lack r1 of HiC reads (--h1)\n"); return 0; } if(asm_opt->hic_reads[0] != NULL && asm_opt->hic_reads[1] != NULL && asm_opt->hic_reads[0]->n != asm_opt->hic_reads[1]->n) { fprintf(stderr, "[ERROR] wrong r1 and r2 of HiC reads (--h1 && --h2)\n"); return 0; } if(asm_opt->hic_enzymes != NULL && asm_opt->hic_enzymes->n == 0) { fprintf(stderr, "[ERROR] wrong HiC enzymes (--enzyme)\n"); return 0; } if(asm_opt->hic_reads[0] != NULL && asm_opt->hic_reads[0]->n == 0) { fprintf(stderr, "[ERROR] wrong r1 of HiC reads (--h1)\n"); return 0; } if(asm_opt->hic_reads[1] != NULL && asm_opt->hic_reads[1]->n == 0) { fprintf(stderr, "[ERROR] wrong r2 of HiC reads (--h2)\n"); return 0; } if(asm_opt->ar != NULL && check_hic_reads(asm_opt->ar, "UL") == 0) return 0; if(asm_opt->ar != NULL && asm_opt->ar->n == 0) { fprintf(stderr, "[ERROR] wrong UL reads (--ul)\n"); return 0; } if(asm_opt->b_low_cov < 0) { fprintf(stderr, "[ERROR] must >= 0 (--b-cov)\n"); return 0; } if(asm_opt->b_high_cov != -1 && asm_opt->b_high_cov < 0) { fprintf(stderr, "[ERROR] must >= 0 (--h-cov)\n"); return 0; } if(asm_opt->m_rate < 0) { fprintf(stderr, "[ERROR] must >= 0 (--m-rate)\n"); return 0; } if(asm_opt->b_high_cov != -1 && asm_opt->b_high_cov <= asm_opt->b_low_cov) { fprintf(stderr, "[ERROR] [--h-cov] must >= [--b-cov]\n"); return 0; } if(asm_opt->purge_simi_thres < 0) { fprintf(stderr, "[ERROR] [-s] must >= 0\n"); return 0; } if(asm_opt->max_kmer_cnt < 0) { fprintf(stderr, "[ERROR] [--max-kocc] must >= 0\n"); return 0; } if(asm_opt->hg_size < -1) { fprintf(stderr, "[ERROR] [--hg-size] wrong genome size\n"); return 0; } return 1; } void get_queries(int argc, char *argv[], ketopt_t* opt, hifiasm_opt_t* asm_opt) { if(opt->ind == argc) { return; } asm_opt->num_reads = argc - opt->ind; asm_opt->read_file_names = (char**)malloc(sizeof(char*)*asm_opt->num_reads); long long i; gzFile dfp; for (i = 0; i < asm_opt->num_reads; i++) { asm_opt->read_file_names[i] = argv[i + opt->ind]; dfp = gzopen(asm_opt->read_file_names[i], "r"); if (dfp == 0) { fprintf(stderr, "[ERROR] Cannot find the input read file: %s\n", asm_opt->read_file_names[i]); exit(0); } gzclose(dfp); } } void get_hic_enzymes(char *argv, enzyme** x, int check_name) { int i, k, pre_i, len = strlen(argv); (*x) = (enzyme*)calloc(1, sizeof(enzyme)); if(len == 0) { (*x)->n = 0; (*x)->l = NULL; (*x)->a = NULL; return; } (*x)->n = 1; for (i = pre_i = 0; i < len; i++) { if(argv[i] == ',') { (*x)->n++; continue; } if(check_name) { if(argv[i] != 'A' && argv[i] != 'C' && argv[i] != 'G' && argv[i] != 'T' && argv[i] != 'a' && argv[i] != 'c' && argv[i] != 'g' && argv[i] != 't' && argv[i] != 'N' && argv[i] != 'n') { (*x)->n = 0; (*x)->l = NULL; (*x)->a = NULL; return; } } } (*x)->l = (int*)calloc((*x)->n, sizeof(int)); (*x)->a = (char**)calloc((*x)->n, sizeof(char*)); for (i = pre_i = k = 0; i < len; i++) { if(argv[i] == ',') { (*x)->l[k] = i - pre_i; (*x)->a[k] = (char*)malloc(sizeof(char)*((*x)->l[k]+1)); memcpy((*x)->a[k], argv + pre_i, (*x)->l[k]); (*x)->a[k][(*x)->l[k]] = '\0'; pre_i = i + 1; k++; } } (*x)->l[k] = i - pre_i; (*x)->a[k] = (char*)malloc(sizeof(char)*((*x)->l[k]+1)); memcpy((*x)->a[k], argv + pre_i, (*x)->l[k]); (*x)->a[k][(*x)->l[k]] = '\0'; } int64_t inter_gsize(char *argv) { int64_t len = strlen(argv); double s; if(len <= 1) return -2; char t = argv[len-1]; if(t != 'k' && t != 'K' && t != 'm' && t != 'M' && t != 'g' && t != 'G') return -2; char *ss=(char*)malloc(len); memcpy(ss, argv, len-1); ss[len-1] = '\0'; s = atof(ss); free(ss); if(t == 'k' || t == 'K') return s*1000; if(t == 'm' || t == 'M') return s*1000000; if(t == 'g' || t == 'G') return s*1000000000; return s; } int CommandLine_process(int argc, char *argv[], hifiasm_opt_t* asm_opt) { ketopt_t opt = KETOPT_INIT; int c; while ((c = ketopt(&opt, argc, argv, 1, "hvt:o:k:w:m:n:r:a:b:z:x:y:p:c:d:M:P:if:D:FN:1:2:3:4:5:l:s:O:eu:", long_options)) >= 0) { if (c == 'h') { Print_H(asm_opt); return 0; } else if (c == 'v' || c == 300) { puts(HA_VERSION); return 0; } else if (c == 'f') asm_opt->bf_shift = atoi(opt.arg); else if (c == 't') asm_opt->thread_num = atoi(opt.arg); else if (c == 'o') asm_opt->output_file_name = opt.arg; else if (c == 'r') asm_opt->number_of_round = atoi(opt.arg); else if (c == 'k') asm_opt->k_mer_length = atoi(opt.arg); else if (c == 'i') asm_opt->load_index_from_disk = 0; else if (c == 'w') asm_opt->mz_win = atoi(opt.arg); else if (c == 'D') asm_opt->high_factor = atof(opt.arg); else if (c == 'F') asm_opt->flag |= HA_F_NO_KMER_FLT; else if (c == 'N') asm_opt->max_n_chain = atoi(opt.arg); else if (c == 'a') asm_opt->clean_round = atoi(opt.arg); else if (c == 'z') asm_opt->adapterLen = atoi(opt.arg); else if (c == 'b') asm_opt->required_read_name = opt.arg; else if (c == 'c') asm_opt->min_cnt = atoi(opt.arg); else if (c == 'd') asm_opt->mid_cnt = atoi(opt.arg); else if (c == '1' || c == 'P') asm_opt->fn_bin_yak[0] = opt.arg; // -P/-M reserved for backward compatibility else if (c == '2' || c == 'M') asm_opt->fn_bin_yak[1] = opt.arg; else if (c == '3') asm_opt->fn_bin_list[0] = opt.arg; else if (c == '4') asm_opt->fn_bin_list[1] = opt.arg; else if (c == '5') asm_opt->fn_bin_poy = opt.arg; else if (c == 'x') asm_opt->max_drop_rate = atof(opt.arg); else if (c == 'y') asm_opt->min_drop_rate = atof(opt.arg); else if (c == 'p') asm_opt->small_pop_bubble_size = atoll(opt.arg); else if (c == 'm') asm_opt->large_pop_bubble_size = atoll(opt.arg); else if (c == 'n') asm_opt->max_short_tip = atoll(opt.arg); else if (c == 'e') asm_opt->flag |= HA_F_BAN_ASSEMBLY; else if (c == 'u') { if(atoll(opt.arg)) { asm_opt->hifi_pst_join = asm_opt->ul_pst_join = 1; } else { asm_opt->hifi_pst_join = asm_opt->ul_pst_join = 0; } } else if (c == 301) asm_opt->flag |= HA_F_VERBOSE_GFA; else if (c == 302) asm_opt->flag |= HA_F_WRITE_PAF; else if (c == 303) asm_opt->flag |= HA_F_WRITE_EC; else if (c == 304) asm_opt->flag |= HA_F_SKIP_TRIOBIN; else if (c == 305) asm_opt->max_ov_diff_ec = atof(opt.arg); else if (c == 306) asm_opt->max_ov_diff_final = atof(opt.arg); else if (c == 307) asm_opt->extract_list = opt.arg; else if (c == 308) asm_opt->extract_iter = atoi(opt.arg); else if (c == 309) { asm_opt->hom_global_coverage = atoi(opt.arg); asm_opt->hom_global_coverage_set = 1; } else if (c == 310) { char* s = NULL; asm_opt->recover_atg_cov_min = strtol(opt.arg, &s, 10); if (*s == ',') asm_opt->recover_atg_cov_max = strtol(s + 1, &s, 10); if(asm_opt->recover_atg_cov_min == -1 || asm_opt->recover_atg_cov_max == -1) { asm_opt->recover_atg_cov_min = asm_opt->recover_atg_cov_max = -1; } } ///else if (c == 311) asm_opt->flag |= HA_F_HIGH_HET; else if (c == 312) asm_opt->bed_inconsist_rate = atoi(opt.arg); else if (c == 313) asm_opt->min_hist_kmer_cnt = atoi(opt.arg); else if (c == 314) get_hic_enzymes(opt.arg, &(asm_opt->hic_reads[0]), 0); else if (c == 315) get_hic_enzymes(opt.arg, &(asm_opt->hic_reads[1]), 0); else if (c == 316) get_hic_enzymes(opt.arg, &(asm_opt->hic_enzymes), 1); else if (c == 317) asm_opt->b_low_cov = atoi(opt.arg); else if (c == 318) asm_opt->b_high_cov = atoi(opt.arg); else if (c == 319) asm_opt->m_rate = atof(opt.arg); else if (c == 320) asm_opt->flag -= HA_F_PARTITION, asm_opt->is_alt = 1; else if (c == 321) asm_opt->trio_flag_occ_thres = atoi(opt.arg); else if (c == 322) asm_opt->seed = atol(opt.arg); else if (c == 323) asm_opt->n_perturb = atoi(opt.arg); else if (c == 324) asm_opt->f_perturb = atof(opt.arg); else if (c == 325) asm_opt->polyploidy = atoi(opt.arg); else if (c == 326) asm_opt->n_weight = atoi(opt.arg); else if (c == 327) asm_opt->misjoin_len = atol(opt.arg); else if (c == 328) asm_opt->pur_global_coverage = atoi(opt.arg); else if (c == 329) asm_opt->flag |= HA_F_FAST; else if (c == 330) asm_opt->dp_e = atof(opt.arg); else if (c == 331) asm_opt->max_kmer_cnt = atol(opt.arg); else if (c == 332) asm_opt->hg_size = inter_gsize(opt.arg); else if (c == 333) get_hic_enzymes(opt.arg, &(asm_opt->ar), 0); else if (c == 334) asm_opt->flag |= HA_F_USKEW; else if (c == 335) asm_opt->kpt_rate = atof(opt.arg); else if (c == 336) asm_opt->ul_error_rate = atof(opt.arg); else if (c == 337) asm_opt->is_dbg_het_cnt = 1; else if (c == 338) asm_opt->max_short_ul_tip = atol(opt.arg); else if (c == 339) asm_opt->is_low_het_ul = 1; else if (c == 340) { asm_opt->trans_base_rate_sec = atof(opt.arg); if(asm_opt->trans_base_rate_sec < 0) asm_opt->is_base_trans = 0; } else if (c == 341) asm_opt->bin_only = 1; else if (c == 342) asm_opt->ul_clean_round = atol(opt.arg); else if (c == 343) asm_opt->prt_dbg_gfa = 1; else if (c == 344) asm_opt->integer_correct_round = atol(opt.arg); else if (c == 345) asm_opt->dbg_ovec_cal = 1; else if (c == 346) asm_opt->max_path_drop_rate = atof(opt.arg); else if (c == 347) asm_opt->min_path_drop_rate = atof(opt.arg); else if (c == 348) asm_opt->trio_cov_het_ovlp = 1; else if (c == 349) asm_opt->ul_min_base = atol(opt.arg); else if (c == 350) asm_opt->self_scaf = 1; else if (c == 351) asm_opt->self_scaf_gap_max = atol(opt.arg); else if (c == 'l') { ///0: disable purge_dup; 1: purge containment; 2: purge overlap asm_opt->purge_level_primary = asm_opt->purge_level_trio = atoi(opt.arg); } else if (c == 's') asm_opt->purge_simi_rate_l2 = asm_opt->purge_simi_rate_l3 = atof(opt.arg); else if (c == 'O') asm_opt->purge_overlap_len = atoll(opt.arg); else if (c == ':') { fprintf(stderr, "[ERROR] missing option argument in \"%s\"\n", argv[opt.i - 1]); return 1; } else if (c == '?') { fprintf(stderr, "[ERROR] unknown option in \"%s\"\n", argv[opt.i - 1]); return 1; } } if(asm_opt->purge_level_primary > 2) asm_opt->purge_simi_thres = asm_opt->purge_simi_rate_l3; else asm_opt->purge_simi_thres = asm_opt->purge_simi_rate_l2; if (argc == opt.ind) { Print_H(asm_opt); return 0; } get_queries(argc, argv, &opt, asm_opt); c = ((asm_opt->ar)?(asm_opt->ul_pst_join):(asm_opt->hifi_pst_join)); if(c) { if((asm_opt->flag&HA_F_BAN_POST_JOIN)) asm_opt->flag-=HA_F_BAN_POST_JOIN; } else { asm_opt->flag |= HA_F_BAN_POST_JOIN; } // fprintf(stderr, "[M::%s::] post join::%u\n", __func__, (uint32_t)(!(asm_opt->flag & HA_F_BAN_POST_JOIN))); // exit(1); return check_option(asm_opt); }