int ctx_exp_abc(int argc, char **argv)
{
size_t i, nthreads = 0, num_repeats = 0, max_AB_dist = 0;
struct MemArgs memargs = MEM_ARGS_INIT;
bool print_failed_contigs = false;
GPathReader tmp_gpfile;
GPathFileBuffer gpfiles;
gpfile_buf_alloc(&gpfiles, 8);
// Arg parsing
char cmd[100];
char shortopts[300];
cmd_long_opts_to_short(longopts, shortopts, sizeof(shortopts));
int c;
// silence error messages from getopt_long
// opterr = 0;
while((c = getopt_long_only(argc, argv, shortopts, longopts, NULL)) != -1) {
cmd_get_longopt_str(longopts, c, cmd, sizeof(cmd));
switch(c) {
case 0: /* flag set */ break;
case 'h': cmd_print_usage(NULL); break;
case 't': cmd_check(!nthreads,cmd); nthreads = cmd_uint32_nonzero(cmd, optarg); break;
case 'm': cmd_mem_args_set_memory(&memargs, optarg); break;
case 'n': cmd_mem_args_set_nkmers(&memargs, optarg); break;
case 'p':
memset(&tmp_gpfile, 0, sizeof(GPathReader));
gpath_reader_open(&tmp_gpfile, optarg);
gpfile_buf_push(&gpfiles, &tmp_gpfile, 1);
break;
case 'N': cmd_check(!num_repeats,cmd); num_repeats = cmd_uint32_nonzero(cmd, optarg); break;
case 'M': cmd_check(!max_AB_dist,cmd); max_AB_dist = cmd_uint32_nonzero(cmd, optarg); break;
case 'P': cmd_check(!print_failed_contigs,cmd); print_failed_contigs = true; break;
case ':': /* BADARG */
case '?': /* BADCH getopt_long has already printed error */
// cmd_print_usage(NULL);
die("`"CMD" exp_abc -h` for help. Bad option: %s", argv[optind-1]);
default: abort();
}
}
// Defaults
if(nthreads == 0) nthreads = DEFAULT_NTHREADS;
if(num_repeats == 0) num_repeats = DEFAULT_NUM_REPEATS;
if(max_AB_dist == 0) max_AB_dist = DEFAULT_MAX_AB_DIST;
if(print_failed_contigs && nthreads != 1) {
warn("--print forces nthreads to be one. soz.");
nthreads = 1;
}
if(optind+1 != argc) cmd_print_usage("Require exactly one input graph file (.ctx)");
const char *ctx_path = argv[optind];
//
// Open Graph file
//
GraphFileReader gfile;
memset(&gfile, 0, sizeof(GraphFileReader));
graph_file_open(&gfile, ctx_path);
size_t ncols = file_filter_into_ncols(&gfile.fltr);
// Check only loading one colour
if(ncols > 1) die("Only implemented for one colour currently");
// Check graph + paths are compatible
graphs_gpaths_compatible(&gfile, 1, gpfiles.b, gpfiles.len, -1);
//
// Decide on memory
//
size_t bits_per_kmer, kmers_in_hash, graph_mem, path_mem, total_mem;
// 1 bit needed per kmer if we need to keep track of kmer usage
bits_per_kmer = sizeof(BinaryKmer)*8 + sizeof(Edges)*8 + sizeof(GPath*)*8 +
ncols;
kmers_in_hash = cmd_get_kmers_in_hash(memargs.mem_to_use,
memargs.mem_to_use_set,
memargs.num_kmers,
memargs.num_kmers_set,
bits_per_kmer,
gfile.num_of_kmers, gfile.num_of_kmers,
false, &graph_mem);
// Paths memory
size_t rem_mem = memargs.mem_to_use - MIN2(memargs.mem_to_use, graph_mem);
path_mem = gpath_reader_mem_req(gpfiles.b, gpfiles.len, ncols, rem_mem, false,
kmers_in_hash, false);
// Shift path store memory from graphs->paths
graph_mem -= sizeof(GPath*)*kmers_in_hash;
path_mem += sizeof(GPath*)*kmers_in_hash;
cmd_print_mem(path_mem, "paths");
total_mem = graph_mem + path_mem;
cmd_check_mem_limit(memargs.mem_to_use, total_mem);
//
// Allocate memory
//
dBGraph db_graph;
db_graph_alloc(&db_graph, gfile.hdr.kmer_size, 1, 1, kmers_in_hash,
DBG_ALLOC_EDGES | DBG_ALLOC_NODE_IN_COL);
// Paths
gpath_reader_alloc_gpstore(gpfiles.b, gpfiles.len, path_mem, false, &db_graph);
// Load the graph
GraphLoadingPrefs gprefs = graph_loading_prefs(&db_graph);
gprefs.empty_colours = true;
graph_load(&gfile, gprefs, NULL);
graph_file_close(&gfile);
hash_table_print_stats(&db_graph.ht);
// Load link files
for(i = 0; i < gpfiles.len; i++) {
gpath_reader_load(&gpfiles.b[i], GPATH_DIE_MISSING_KMERS, &db_graph);
gpath_reader_close(&gpfiles.b[i]);
}
gpfile_buf_dealloc(&gpfiles);
status("\n");
status("Test 1: Priming region A->B (n: %zu max_AB_dist: %zu)",
num_repeats, max_AB_dist);
run_exp_abc(&db_graph, true, nthreads, num_repeats,
max_AB_dist, print_failed_contigs);
status("\n");
status("Test 2: Trying to traverse A->B (n: %zu max_AB_dist: %zu)",
num_repeats, max_AB_dist);
run_exp_abc(&db_graph, false, nthreads, num_repeats,
max_AB_dist, print_failed_contigs);
db_graph_dealloc(&db_graph);
return EXIT_SUCCESS;
}
int ctx_vcfcov(int argc, char **argv)
{
struct MemArgs memargs = MEM_ARGS_INIT;
const char *out_path = NULL, *out_type = NULL;
uint32_t max_allele_len = 0, max_gt_vars = 0;
char *ref_path = NULL;
bool low_mem = false;
// Arg parsing
char cmd[100];
char shortopts[300];
cmd_long_opts_to_short(longopts, shortopts, sizeof(shortopts));
int c;
size_t i;
// silence error messages from getopt_long
// opterr = 0;
while((c = getopt_long_only(argc, argv, shortopts, longopts, NULL)) != -1) {
cmd_get_longopt_str(longopts, c, cmd, sizeof(cmd));
switch(c) {
case 0: /* flag set */ break;
case 'h': cmd_print_usage(NULL); break;
case 'o': cmd_check(!out_path, cmd); out_path = optarg; break;
case 'O': cmd_check(!out_type, cmd); out_type = optarg; break;
case 'f': cmd_check(!futil_get_force(), cmd); futil_set_force(true); break;
case 'm': cmd_mem_args_set_memory(&memargs, optarg); break;
case 'n': cmd_mem_args_set_nkmers(&memargs, optarg); break;
case 'r': cmd_check(!ref_path, cmd); ref_path = optarg; break;
case 'L': cmd_check(!max_allele_len,cmd); max_allele_len = cmd_uint32(cmd,optarg); break;
case 'N': cmd_check(!max_gt_vars,cmd); max_gt_vars = cmd_uint32(cmd,optarg); break;
case 'M': cmd_check(!low_mem, cmd); low_mem = true; break;
case ':': /* BADARG */
case '?': /* BADCH getopt_long has already printed error */
// cmd_print_usage(NULL);
die("`"CMD" "SUBCMD" -h` for help. Bad option: %s", argv[optind-1]);
default: abort();
}
}
// Defaults for unset values
if(out_path == NULL) out_path = "-";
if(ref_path == NULL) cmd_print_usage("Require a reference (-r,--ref <ref.fa>)");
if(optind+2 > argc) cmd_print_usage("Require VCF and graph files");
if(!max_allele_len) max_allele_len = DEFAULT_MAX_ALLELE_LEN;
if(!max_gt_vars) max_gt_vars = DEFAULT_MAX_GT_VARS;
status("[vcfcov] max allele length: %u; max number of variants: %u",
max_allele_len, max_gt_vars);
// open ref
// index fasta with: samtools faidx ref.fa
faidx_t *fai = fai_load(ref_path);
if(fai == NULL) die("Cannot load ref index: %s / %s.fai", ref_path, ref_path);
// Open input VCF file
const char *vcf_path = argv[optind++];
htsFile *vcffh = hts_open(vcf_path, "r");
if(vcffh == NULL) die("Cannot open VCF file: %s", vcf_path);
bcf_hdr_t *vcfhdr = bcf_hdr_read(vcffh);
if(vcfhdr == NULL) die("Cannot read VCF header: %s", vcf_path);
// Test we can close and reopen files
if(low_mem) {
if((vcffh = hts_open(vcf_path, "r")) == NULL)
die("Cannot re-open VCF file: %s", vcf_path);
if((vcfhdr = bcf_hdr_read(vcffh)) == NULL)
die("Cannot re-read VCF header: %s", vcf_path);
}
//
// Open graph files
//
const size_t num_gfiles = argc - optind;
char **graph_paths = argv + optind;
ctx_assert(num_gfiles > 0);
GraphFileReader *gfiles = ctx_calloc(num_gfiles, sizeof(GraphFileReader));
size_t ncols, ctx_max_kmers = 0, ctx_sum_kmers = 0;
ncols = graph_files_open(graph_paths, gfiles, num_gfiles,
&ctx_max_kmers, &ctx_sum_kmers);
// Check graph + paths are compatible
graphs_gpaths_compatible(gfiles, num_gfiles, NULL, 0, -1);
//
// Decide on memory
//
size_t bits_per_kmer, kmers_in_hash, graph_mem;
bits_per_kmer = sizeof(BinaryKmer)*8 + sizeof(Covg)*8 * ncols;
kmers_in_hash = cmd_get_kmers_in_hash(memargs.mem_to_use,
memargs.mem_to_use_set,
memargs.num_kmers,
memargs.num_kmers_set,
bits_per_kmer,
low_mem ? -1 : (int64_t)ctx_max_kmers,
ctx_sum_kmers,
true, &graph_mem);
cmd_check_mem_limit(memargs.mem_to_use, graph_mem);
//
// Open output file
//
// v=>vcf, z=>compressed vcf, b=>bcf, bu=>uncompressed bcf
int mode = vcf_misc_get_outtype(out_type, out_path);
futil_create_output(out_path);
htsFile *outfh = hts_open(out_path, modes_htslib[mode]);
status("[vcfcov] Output format: %s", hsmodes_htslib[mode]);
// Allocate memory
dBGraph db_graph;
db_graph_alloc(&db_graph, gfiles[0].hdr.kmer_size, ncols, 1, kmers_in_hash,
DBG_ALLOC_COVGS);
//
// Set up tag names
//
// *R => ref, *A => alt
sprintf(kcov_ref_tag, "K%zuR", db_graph.kmer_size); // mean coverage
sprintf(kcov_alt_tag, "K%zuA", db_graph.kmer_size);
// #SAMPLE=<ID=...,K29KCOV=...,K29NK=...,K29RLK>
// - K29_kcov is empirical kmer coverage
// - K29_nkmers is the number of kmers in the sample
// - mean_read_length is the mean read length in bases
char sample_kcov_tag[20], sample_nk_tag[20], sample_rlk_tag[20];
sprintf(sample_kcov_tag, "K%zu_kcov", db_graph.kmer_size); // mean coverage
sprintf(sample_nk_tag, "K%zu_nkmers", db_graph.kmer_size);
sprintf(sample_rlk_tag, "mean_read_length");
//
// Load kmers if we are using --low-mem
//
VcfCovStats st;
memset(&st, 0, sizeof(st));
VcfCovPrefs prefs = {.kcov_ref_tag = kcov_ref_tag,
.kcov_alt_tag = kcov_alt_tag,
.max_allele_len = max_allele_len,
.max_gt_vars = max_gt_vars,
.load_kmers_only = false};
if(low_mem)
{
status("[vcfcov] Loading kmers from VCF+ref");
prefs.load_kmers_only = true;
vcfcov_file(vcffh, vcfhdr, NULL, NULL, vcf_path, fai,
NULL, &prefs, &st, &db_graph);
// Close files
hts_close(vcffh);
bcf_hdr_destroy(vcfhdr);
// Re-open files
if((vcffh = hts_open(vcf_path, "r")) == NULL)
die("Cannot re-open VCF file: %s", vcf_path);
if((vcfhdr = bcf_hdr_read(vcffh)) == NULL)
die("Cannot re-read VCF header: %s", vcf_path);
prefs.load_kmers_only = false;
}
//
// Load graphs
//
GraphLoadingStats gstats;
memset(&gstats, 0, sizeof(gstats));
GraphLoadingPrefs gprefs = graph_loading_prefs(&db_graph);
gprefs.must_exist_in_graph = low_mem;
for(i = 0; i < num_gfiles; i++) {
graph_load(&gfiles[i], gprefs, &gstats);
graph_file_close(&gfiles[i]);
}
ctx_free(gfiles);
hash_table_print_stats(&db_graph.ht);
//
// Set up VCF header / graph matchup
//
size_t *samplehdrids = ctx_malloc(db_graph.num_of_cols * sizeof(size_t));
// Add samples to vcf header
bcf_hdr_t *outhdr = bcf_hdr_dup(vcfhdr);
bcf_hrec_t *hrec;
int sid;
char hdrstr[200];
for(i = 0; i < db_graph.num_of_cols; i++) {
char *sname = db_graph.ginfo[i].sample_name.b;
if((sid = bcf_hdr_id2int(outhdr, BCF_DT_SAMPLE, sname)) < 0) {
bcf_hdr_add_sample(outhdr, sname);
sid = bcf_hdr_id2int(outhdr, BCF_DT_SAMPLE, sname);
}
samplehdrids[i] = sid;
// Add SAMPLE field
hrec = bcf_hdr_get_hrec(outhdr, BCF_HL_STR, "ID", sname, "SAMPLE");
if(hrec == NULL) {
sprintf(hdrstr, "##SAMPLE=<ID=%s,%s=%"PRIu64",%s=%"PRIu64",%s=%zu>", sname,
sample_kcov_tag,
gstats.nkmers[i] ? gstats.sumcov[i] / gstats.nkmers[i] : 0,
sample_nk_tag, gstats.nkmers[i],
sample_rlk_tag, (size_t)db_graph.ginfo[i].mean_read_length);
bcf_hdr_append(outhdr, hdrstr);
}
else {
// mean kcovg
sprintf(hdrstr, "%"PRIu64, gstats.sumcov[i] / gstats.nkmers[i]);
vcf_misc_add_update_hrec(hrec, sample_kcov_tag, hdrstr);
// num kmers
sprintf(hdrstr, "%"PRIu64, gstats.nkmers[i]);
vcf_misc_add_update_hrec(hrec, sample_nk_tag, hdrstr);
// mean read length in kmers
sprintf(hdrstr, "%zu", (size_t)db_graph.ginfo[i].mean_read_length);
vcf_misc_add_update_hrec(hrec, sample_rlk_tag, hdrstr);
}
status("[vcfcov] Colour %zu: %s [VCF column %zu]", i, sname, samplehdrids[i]);
}
// Add genotype format fields
// One field per alternative allele
sprintf(hdrstr, "##FORMAT=<ID=%s,Number=A,Type=Integer,"
"Description=\"Coverage on ref (k=%zu): sum(kmer_covs) / exp_num_kmers\">\n",
kcov_ref_tag, db_graph.kmer_size);
bcf_hdr_append(outhdr, hdrstr);
sprintf(hdrstr, "##FORMAT=<ID=%s,Number=A,Type=Integer,"
"Description=\"Coverage on alt (k=%zu): sum(kmer_covs) / exp_num_kmers\">\n",
kcov_alt_tag, db_graph.kmer_size);
bcf_hdr_append(outhdr, hdrstr);
bcf_hdr_set_version(outhdr, "VCFv4.2");
// Add command string to header
vcf_misc_hdr_add_cmd(outhdr, cmd_get_cmdline(), cmd_get_cwd());
if(bcf_hdr_write(outfh, outhdr) != 0)
die("Cannot write header to: %s", futil_outpath_str(out_path));
status("[vcfcov] Reading %s and adding coverage", vcf_path);
// Reset stats and get coverage
memset(&st, 0, sizeof(st));
vcfcov_file(vcffh, vcfhdr, outfh, outhdr, vcf_path, fai,
samplehdrids, &prefs, &st, &db_graph);
// Print statistics
char ns0[50], ns1[50];
status("[vcfcov] Read %s VCF lines", ulong_to_str(st.nvcf_lines, ns0));
status("[vcfcov] Read %s ALTs", ulong_to_str(st.nalts_read, ns0));
status("[vcfcov] Used %s kmers", ulong_to_str(st.ngt_kmers, ns0));
status("[vcfcov] ALTs used: %s / %s (%.2f%%)",
ulong_to_str(st.nalts_loaded, ns0), ulong_to_str(st.nalts_read, ns1),
st.nalts_read ? (100.0*st.nalts_loaded) / st.nalts_read : 0.0);
status("[vcfcov] ALTs too long (>%ubp): %s / %s (%.2f%%)", max_allele_len,
ulong_to_str(st.nalts_too_long, ns0), ulong_to_str(st.nalts_read, ns1),
st.nalts_read ? (100.0*st.nalts_too_long) / st.nalts_read : 0.0);
status("[vcfcov] ALTs too dense (>%u within %zubp): %s / %s (%.2f%%)",
max_gt_vars, db_graph.kmer_size,
ulong_to_str(st.nalts_no_covg, ns0), ulong_to_str(st.nalts_read, ns1),
st.nalts_read ? (100.0*st.nalts_no_covg) / st.nalts_read : 0.0);
status("[vcfcov] ALTs printed with coverage: %s / %s (%.2f%%)",
ulong_to_str(st.nalts_with_covg, ns0), ulong_to_str(st.nalts_read, ns1),
st.nalts_read ? (100.0*st.nalts_with_covg) / st.nalts_read : 0.0);
status("[vcfcov] Saved to: %s\n", out_path);
ctx_free(samplehdrids);
graph_loading_stats_destroy(&gstats);
bcf_hdr_destroy(vcfhdr);
bcf_hdr_destroy(outhdr);
hts_close(vcffh);
hts_close(outfh);
fai_destroy(fai);
db_graph_dealloc(&db_graph);
return EXIT_SUCCESS;
}
