static void gpath_save_thread(void *arg)
{
GPathSaver *wrkr = (GPathSaver*)arg;
const dBGraph *db_graph = wrkr->db_graph;
GPathSubset subset;
StrBuf sbuf;
gpath_subset_alloc(&subset);
gpath_subset_init(&subset, &wrkr->db_graph->gpstore.gpset);
strbuf_alloc(&sbuf, 2 * DEFAULT_IO_BUFSIZE);
dBNodeBuffer nbuf;
SizeBuffer jposbuf;
db_node_buf_alloc(&nbuf, 1024);
size_buf_alloc(&jposbuf, 256);
HASH_ITERATE_PART(&db_graph->ht, wrkr->threadid, wrkr->nthreads,
_gpath_gzsave_node,
&sbuf, &subset,
wrkr->save_seq ? &nbuf : NULL, wrkr->save_seq ? &jposbuf : NULL,
wrkr->gzout, wrkr->outlock,
db_graph);
_gpath_save_flush(wrkr->gzout, &sbuf, wrkr->outlock);
db_node_buf_dealloc(&nbuf);
size_buf_dealloc(&jposbuf);
gpath_subset_dealloc(&subset);
strbuf_dealloc(&sbuf);
}
char *
strbuf_double(idnconv_strbuf_t *buf) {
/*
* Double the size of the buffer of BUF.
*/
return (strbuf_alloc(buf, buf->size * 2));
}
// Returns number of types.
int load_hla_csv(const char *path, char ***bools_ptr, int num_rows)
{
assert(num_rows > 0);
StrBuf line;
strbuf_alloc(&line, 1024);
FILE *fh = fopen(path, "r");
if(fh == NULL) die("Cannot open file: %s.", path);
if(strbuf_readline(&line, fh) == 0) die("Empty CSV file: %s.", path);
int num_types = count_char(line.b, ',');
char **bools = my_malloc(sizeof(char*) * num_rows, __FILE__, __LINE__);
char *data = my_malloc(sizeof(char) * num_rows * (num_types+1), __FILE__, __LINE__);
printf("Number of rows: %i.\n",num_rows);
int i;
for(i = 0; i < num_rows && strbuf_reset_readline(&line, fh); i++)
{
strbuf_chomp(&line);
bools[i] = data + i * (num_types+1);
load_comma_bool_line(line.b, bools[i], num_types);
bools[i][num_types] = '\0';
}
if(i < num_rows) die("Not enough rows in CSV file: %s.", path);
fclose(fh);
strbuf_dealloc(&line);
*bools_ptr = bools;
return num_types;
}
char *
strbuf_copy(idnconv_strbuf_t *buf, const char *str) {
/*
* Copy STR to BUF.
*/
size_t len = strlen(str);
if (strbuf_alloc(buf, len + 1) == NULL)
return (NULL);
strcpy(buf->str, str);
return (buf->str);
}
CallDecomp* call_decomp_init(htsFile *vcffh, bcf_hdr_t *vcfhdr)
{
CallDecomp *dc = ctx_calloc(1, sizeof(CallDecomp));
dc->nw_aligner = needleman_wunsch_new();
dc->aln = alignment_create(1024);
dc->scoring = ctx_calloc(1, sizeof(dc->scoring[0]));
scoring_system_default(dc->scoring);
dc->vcffh = vcffh;
dc->vcfhdr = vcfhdr;
dc->v = bcf_init();
strbuf_alloc(&dc->sbuf, 256);
return dc;
}
char *
strbuf_append(idnconv_strbuf_t *buf, const char *str) {
/*
* Append STR to the end of BUF.
*/
size_t len1 = strlen(buf->str);
size_t len2 = strlen(str);
char *p;
#define MARGIN 50
p = strbuf_alloc(buf, len1 + len2 + 1 + MARGIN);
if (p != NULL)
strcpy(buf->str + len1, str);
return (p);
}
void vcf_misc_hdr_add_cmd(bcf_hdr_t *hdr, const char *cmdline, const char *cwd)
{
char keystr[8], timestr[100];
time_t tnow;
time(&tnow);
strftime(timestr, sizeof(timestr), "%Y%m%d-%H:%M:%S", localtime(&tnow));
StrBuf sbuf;
strbuf_alloc(&sbuf, 1024);
strbuf_sprintf(&sbuf, "##mccortex_%s=<prev=\"NULL\",cmd=\"%s\",cwd=\"%s\","
"datetime=\"%s\",version="CTX_VERSION">\n",
hex_rand_str(keystr, sizeof(keystr)),
cmdline, cwd, timestr);
bcf_hdr_append(hdr, sbuf.b);
strbuf_dealloc(&sbuf);
}
static cJSON* read_input_header(gzFile gzin)
{
cJSON *json;
StrBuf hdrstr;
strbuf_alloc(&hdrstr, 1024);
json_hdr_read(NULL, gzin, input_path, &hdrstr);
json = cJSON_Parse(hdrstr.b);
if(json == NULL) die("Invalid JSON header: %s", input_path);
// Check we can handle the kmer size
kmer_size = json_hdr_get_kmer_size(json, input_path);
db_graph_check_kmer_size(kmer_size, input_path);
strbuf_dealloc(&hdrstr);
return json;
}
void test_graph_crawler()
{
test_status("Testing graph crawler...");
// Construct 1 colour graph with kmer-size=11
dBGraph graph;
const size_t kmer_size = 11, ncols = 3;
db_graph_alloc(&graph, kmer_size, ncols, 1, 2048,
DBG_ALLOC_EDGES | DBG_ALLOC_NODE_IN_COL | DBG_ALLOC_BKTLOCKS);
char graphseq[3][77] =
// < X X X...............
{"GTTCCAGAGCGGAGGTCTCCCAACAACATGGTATAAGTTGTCTAGCCCCGGTTCGCGCGGGTACTTCTTACAGCGC",
"GTTCCAGAGCGGAGGTCTCCCAACAACTTGGTATAAGTTGTCTAGTCCCGGTTCGCGCGGCATTTCAGCATTGTTA",
"GTTCCAGAGCGCGACAGAGTGCATATCACGCTAAGCACAGCCCTCTTCTATCTGCTTTTAAATGGATCAATAATCG"};
build_graph_from_str_mt(&graph, 0, graphseq[0], strlen(graphseq[0]));
build_graph_from_str_mt(&graph, 1, graphseq[1], strlen(graphseq[1]));
build_graph_from_str_mt(&graph, 2, graphseq[2], strlen(graphseq[2]));
// Crawl graph
GraphCrawler crawler;
graph_crawler_alloc(&crawler, &graph);
dBNode node = db_graph_find_str(&graph, graphseq[0]);
dBNode next_node = db_graph_find_str(&graph, graphseq[0]+1);
TASSERT(node.key != HASH_NOT_FOUND);
TASSERT(next_node.key != HASH_NOT_FOUND);
BinaryKmer bkey = db_node_get_bkmer(&graph, node.key);
Edges edges = db_node_get_edges(&graph, node.key, 0);
dBNode next_nodes[4];
Nucleotide next_nucs[4];
size_t i, p, num_next, next_idx;
num_next = db_graph_next_nodes(&graph, bkey, node.orient, edges,
next_nodes, next_nucs);
next_idx = 0;
while(next_idx < num_next && !db_nodes_are_equal(next_nodes[next_idx],next_node))
next_idx++;
TASSERT(next_idx < num_next && db_nodes_are_equal(next_nodes[next_idx],next_node));
// Crawl in all colours
graph_crawler_fetch(&crawler, node, next_nodes, next_idx, num_next,
NULL, graph.num_of_cols, NULL, NULL, NULL);
TASSERT2(crawler.num_paths == 2, "crawler.num_paths: %u", crawler.num_paths);
// Fetch paths
dBNodeBuffer nbuf;
db_node_buf_alloc(&nbuf, 16);
StrBuf sbuf;
strbuf_alloc(&sbuf, 128);
for(p = 0; p < crawler.num_paths; p++) {
db_node_buf_reset(&nbuf);
graph_crawler_get_path_nodes(&crawler, p, &nbuf);
strbuf_ensure_capacity(&sbuf, nbuf.len+graph.kmer_size);
sbuf.end = db_nodes_to_str(nbuf.b, nbuf.len, &graph, sbuf.b);
for(i = 0; i < 3 && strcmp(graphseq[i]+1,sbuf.b) != 0; i++) {}
TASSERT2(i < 3, "seq: %s", sbuf.b);
TASSERT2(sbuf.end == 75, "sbuf.end: %zu", sbuf.end);
TASSERT2(nbuf.len == 65, "nbuf.len: %zu", nbuf.len);
}
strbuf_dealloc(&sbuf);
db_node_buf_dealloc(&nbuf);
graph_crawler_dealloc(&crawler);
db_graph_dealloc(&graph);
}
void graph_info_alloc(GraphInfo *ginfo)
{
strbuf_alloc(&ginfo->sample_name, 256);
error_cleaning_alloc(&ginfo->cleaning);
graph_info_init(ginfo);
}
static void error_cleaning_alloc(ErrorCleaning *ec)
{
strbuf_alloc(&ec->intersection_name, 256);
error_cleaning_init(ec);
}
static void parse_entries(gzFile gzin, FILE *fout)
{
CallFileEntry centry;
call_file_entry_alloc(¢ry);
ChromPosBuffer chrposbuf;
chrompos_buf_alloc(&chrposbuf, 32);
StrBuf tmpbuf, flank3pbuf;
strbuf_alloc(&tmpbuf, 1024);
strbuf_alloc(&flank3pbuf, 1024);
const char *flank5p, *flank3p;
size_t flank5p_len, flank3p_len;
size_t cpy_flnk_5p, cpy_flnk_3p;
const read_t *chrom = NULL;
size_t ref_start = 0, ref_end = 0;
bool mapped = false, fw_strand = false;
const char **genotypes = NULL;
if(!input_bubble_format)
genotypes = ctx_calloc(num_samples, sizeof(char*));
for(; call_file_read(gzin, input_path, ¢ry); num_entries_read++)
{
size_t nlines = call_file_num_lines(¢ry);
ctx_assert2(!(nlines&1) && nlines >= 6, "Too few lines: %zu", nlines);
flank5p = call_file_get_line(¢ry,1);
flank5p_len = call_file_line_len(¢ry,1);
cpy_flnk_5p = cpy_flnk_3p = 0;
// Read a corresponding SAM entry
if(input_bubble_format)
{
// Trim down alleles, add to 3p flank
bubble_trim_alleles(¢ry, &flank3pbuf);
flank3p = flank3pbuf.b;
flank3p_len = flank3pbuf.end;
mapped = sam_fetch_coords(¢ry, flank5p, flank5p_len, flank3p, flank3p_len,
&cpy_flnk_5p, &cpy_flnk_3p,
&chrom, &ref_start, &ref_end, &fw_strand);
}
else {
flank3p = call_file_get_line(¢ry, 3);
flank3p_len = call_file_line_len(¢ry, 3);
mapped = brkpnt_fetch_coords(¢ry, &chrposbuf,
&chrom, &ref_start, &ref_end, &fw_strand,
&cpy_flnk_5p, &cpy_flnk_3p);
}
if(mapped)
{
// Get call id
const char *hdrline = call_file_get_line(¢ry, 0);
char callid[100];
int r = get_callid_str(hdrline, input_bubble_format, callid, sizeof(callid));
if(r == -1) die("Poorly formatted: %s", hdrline);
if(r == -2) die("Call id string is too long: %s", hdrline);
align_entry(¢ry, callid, flank5p, flank5p_len, flank3p, flank3p_len,
cpy_flnk_5p, cpy_flnk_3p,
chrom, ref_start, ref_end, fw_strand,
&tmpbuf, genotypes,
fout);
}
}
ctx_free(genotypes);
call_file_entry_dealloc(¢ry);
chrompos_buf_dealloc(&chrposbuf);
strbuf_dealloc(&tmpbuf);
strbuf_dealloc(&flank3pbuf);
}
int main(int argc, char **argv)
{
// compiler complains about unused function without these linese
(void)kh_clear_ghash;
(void)kh_del_ghash;
if(argc < 2) print_usage(usage, NULL);
char swap_alleles = 0;
int c;
while((c = getopt(argc, argv, "s")) >= 0) {
switch (c) {
case 's': swap_alleles = 1; break;
default: die("Unknown option: %c", c);
}
}
if(optind == argc) print_usage(usage, "Not enough arguments");
char *inputpath = argv[optind];
char **refpaths = argv + optind + 1;
size_t num_refs = argc - optind - 1;
gzFile gzin = gzopen(inputpath, "r");
if(gzin == NULL) die("Cannot read file: %s", inputpath);
size_t i, nchroms = 0, capacity = 1024;
khash_t(ghash) *genome = kh_init(ghash);
read_t *reads = malloc(capacity * sizeof(read_t)), *r;
int hret;
khiter_t k;
for(i = 0; i < num_refs; i++) {
fprintf(stderr, "Loading %s\n", refpaths[i]);
load_reads(refpaths[i], &reads, &capacity, &nchroms);
}
if(num_refs == 0) {
fprintf(stderr, "Loading from stdin\n");
load_reads("-", &reads, &capacity, &nchroms);
}
if(nchroms == 0) die("No chromosomes loaded");
for(i = 0; i < nchroms; i++) {
r = reads + i;
fprintf(stderr, "Loaded: '%s'\n", r->name.b);
k = kh_put(ghash, genome, r->name.b, &hret);
if(hret == 0) warn("Duplicate read name (taking first): %s", r->name.b);
else kh_value(genome, k) = r;
}
// Now read VCF
StrBuf line;
strbuf_alloc(&line, 1024);
char *fields[9];
char *chr;
int pos, reflen, altlen;
while(strbuf_reset_gzreadline(&line, gzin) > 0)
{
if(line.b[0] == '#') fputs(line.b, stdout);
else
{
strbuf_chomp(&line);
vcf_columns(line.b, fields);
fields[1][-1] = fields[2][-1] = '\0';
chr = line.b;
pos = atoi(fields[1])-1;
k = kh_get(ghash, genome, chr);
r = kh_value(genome, k);
fields[1][-1] = fields[2][-1] = '\t';
reflen = fields[4] - fields[3] - 1;
altlen = fields[5] - fields[4] - 1;
if(k == kh_end(genome)) warn("Cannot find chrom: %s", chr);
else if(pos < 0) warn("Bad line: %s\n", line.b);
else if((reflen == 1 && altlen == 1) || fields[3][0] == fields[4][0])
{
if((unsigned)pos + reflen <= r->seq.end &&
strncasecmp(r->seq.b+pos,fields[3],reflen) == 0)
{
fputs(line.b, stdout);
fputc('\n', stdout);
}
else if(swap_alleles && (unsigned)pos + altlen <= r->seq.end &&
strncasecmp(r->seq.b+pos,fields[4],altlen) == 0)
{
// swap alleles
char tmp[altlen], *ref = fields[3], *alt = fields[4];
memcpy(tmp, alt, altlen);
memmove(ref+altlen+1, ref, reflen);
memcpy(ref, tmp, altlen);
ref[altlen] = '\t';
fputs(line.b, stdout);
fputc('\n', stdout);
}
// else printf("FAIL0\n");
}
// else printf("FAIL1\n");
}
}
kh_destroy(ghash, genome);
strbuf_dealloc(&line);
gzclose(gzin);
for(i = 0; i < nchroms; i++) seq_read_dealloc(reads+i);
free(reads);
fprintf(stderr, " Done.\n");
return 0;
}
BubbleCaller* bubble_callers_new(size_t num_callers,
BubbleCallingPrefs prefs,
gzFile gzout,
const dBGraph *db_graph)
{
ctx_assert(num_callers > 0);
// Max usage is 4 * max_allele_len * cols
size_t i;
size_t max_path_len = MAX2(prefs.max_flank_len, prefs.max_allele_len);
BubbleCaller *callers = ctx_malloc(num_callers * sizeof(BubbleCaller));
pthread_mutex_t *out_lock = ctx_malloc(sizeof(pthread_mutex_t));
if(pthread_mutex_init(out_lock, NULL) != 0) die("mutex init failed");
size_t *num_bubbles_ptr = ctx_calloc(1, sizeof(size_t));
for(i = 0; i < num_callers; i++)
{
BubbleCaller tmp = {.threadid = i, .nthreads = num_callers,
.haploid_seen = ctx_calloc(1+prefs.num_haploid, sizeof(bool)),
.num_bubbles_ptr = num_bubbles_ptr,
.prefs = prefs,
.db_graph = db_graph, .gzout = gzout,
.out_lock = out_lock};
memcpy(&callers[i], &tmp, sizeof(BubbleCaller));
// First two buffers don't actually need to grow
db_node_buf_alloc(&callers[i].flank5p, prefs.max_flank_len);
db_node_buf_alloc(&callers[i].pathbuf, max_path_len);
graph_walker_alloc(&callers[i].wlk, db_graph);
rpt_walker_alloc(&callers[i].rptwlk, db_graph->ht.capacity, 22); // 4MB
graph_cache_alloc(&callers[i].cache, db_graph);
cache_stepptr_buf_alloc(&callers[i].spp_forward, 1024);
cache_stepptr_buf_alloc(&callers[i].spp_reverse, 1024);
strbuf_alloc(&callers[i].output_buf, 2048);
}
return callers;
}
void bubble_callers_destroy(BubbleCaller *callers, size_t num_callers)
{
ctx_assert(num_callers > 0);
size_t i;
for(i = 0; i < num_callers; i++)
{
ctx_free(callers[i].haploid_seen);
db_node_buf_dealloc(&callers[i].flank5p);
db_node_buf_dealloc(&callers[i].pathbuf);
rpt_walker_dealloc(&callers[i].rptwlk);
graph_walker_dealloc(&callers[i].wlk);
graph_cache_dealloc(&callers[i].cache);
cache_stepptr_buf_dealloc(&callers[i].spp_forward);
cache_stepptr_buf_dealloc(&callers[i].spp_reverse);
strbuf_dealloc(&callers[i].output_buf);
}
pthread_mutex_destroy(callers[0].out_lock);
ctx_free(callers[0].out_lock);
ctx_free(callers[0].num_bubbles_ptr);
ctx_free(callers);
}
// Load each sequence into a separate colour
static void test_bubbles(dBGraph *graph, const char **seqs, size_t nseqs,
const char *flank5p, const char *flank3p,
const char **alleles, size_t nalleles)
{
db_graph_reset(graph);
TASSERT(graph->num_of_cols >= nseqs);
size_t i;
for(i = 0; i < nseqs; i++)
build_graph_from_str_mt(graph, i, seqs[i], strlen(seqs[i]), false);
graph->num_of_cols_used = MAX2(graph->num_of_cols_used, 1);
StrBuf sbuf;
dBNodeBuffer nbuf;
strbuf_alloc(&sbuf, 128);
db_node_buf_alloc(&nbuf, 128);
BubbleCallingPrefs prefs = {.max_allele_len = 100, .max_flank_len = 100,
.haploid_cols = NULL, .nhaploid_cols = 0,
.remove_serial_bubbles = true};
BubbleCaller *caller = bubble_callers_new(1, &prefs, NULL, graph);
_call_bubble(caller, flank5p, flank3p, alleles, nalleles, &nbuf, &sbuf);
strbuf_dealloc(&sbuf);
db_node_buf_dealloc(&nbuf);
bubble_callers_destroy(caller, 1);
}
void test_bubble_caller()
{
test_status("Testing bubble calling...");
// Construct 1 colour graph with kmer-size=11
dBGraph graph;
const size_t kmer_size = 11, ncols = 3;
// Create graph
db_graph_alloc(&graph, kmer_size, ncols, 1, 2000,
DBG_ALLOC_EDGES | DBG_ALLOC_NODE_IN_COL | DBG_ALLOC_BKTLOCKS);
// mutations: x
const char *seqs0[] = {"AGGGATAAAACTCTGTACTGGATCTCCCT",
"AGGGATAAAACTCTcTACTGGATCTCCCT"};
const char flank5p0[] = "AGGGATAAAACTCT";
const char flank3p0[] = "TACTGGATCTCCCT";
const char *alleles0[] = {"ATAAAACTCTGTACTGGATCT", "ATAAAACTCTcTACTGGATCT"};
test_bubbles(&graph, seqs0, 2, flank5p0, flank3p0, alleles0, 2);
// mutations: x y
const char *seqs1[] = {"CCCGTAGGTAAGGGCGTTAGTGCAAGGCCACATTGGGACACGAGTTGATA",
"CCCGTAGGTAAGtGCGTTAGTGCAAGGCCACATTGGGACACGAGTTGATA",
"CCCGTAGGTAAGGGCGTTAGTGCAAGGCCACtTTGGGACACGAGTTGATA"};
// forwards
const char flank5p1a[] = "CCCGTAGGTAAG";
const char flank3p1a[] = "GCGTTAGTGCAAGGCCAC";
const char *alleles1a[] = {"CGTAGGTAAGGGCGTTAGTGC", "CGTAGGTAAGtGCGTTAGTGC"};
const char flank5p1b[] = "GCGTTAGTGCAAGGCCAC";
const char flank3p1b[] = "TTGGGACACGAGTTGATA";
const char *alleles1b[] = {"GCAAGGCCACATTGGGACACG", "GCAAGGCCACtTTGGGACACG"};
test_bubbles(&graph, seqs1, 3, flank5p1a, flank3p1a, alleles1a, 2);
test_bubbles(&graph, seqs1, 3, flank5p1b, flank3p1b, alleles1b, 2);
// reverse
// mutations: y x
// TATCAACTCGTGTCCCAATGTGGCCTTGCACTAACGCCCTTACCTACGGG
// TATCAACTCGTGTCCCAATGTGGCCTTGCACTAACGCaCTTACCTACGGG
// TATCAACTCGTGTCCCAAaGTGGCCTTGCACTAACGCCCTTACCTACGGG
//
const char flank5p1c[] = "GTGGCCTTGCACTAACGC";
const char flank3p1c[] = "CTTACCTACGGG";
const char *alleles1c[] = {"GCACTAACGCCCTTACCTACG", "GCACTAACGCaCTTACCTACG"};
const char flank5p1d[] = "TATCAACTCGTGTCCCAA";
const char flank3p1d[] = "GTGGCCTTGCACTAACGC";
const char *alleles1d[] = {"CGTGTCCCAATGTGGCCTTGC", "CGTGTCCCAAaGTGGCCTTGC"};
test_bubbles(&graph, seqs1, 3, flank5p1c, flank3p1c, alleles1c, 2);
test_bubbles(&graph, seqs1, 3, flank5p1d, flank3p1d, alleles1d, 2);
db_graph_dealloc(&graph);
}
static bcf_hdr_t* make_vcf_hdr(cJSON *json, const char *in_path,
bool is_breakpoint, size_t kmer_size,
char const*const* ref_paths, size_t nref_paths,
read_t *chroms, size_t nchroms)
{
ctx_assert(json != NULL);
StrBuf hdrbuf;
strbuf_alloc(&hdrbuf, 1024);
char datestr[9];
time_t date = time(NULL);
strftime(datestr, 9, "%Y%m%d", localtime(&date));
strbuf_append_str(&hdrbuf, "##fileformat=VCFv4.2\n##fileDate=");
strbuf_append_str(&hdrbuf, datestr);
strbuf_append_str(&hdrbuf, "\n");
// Print commands used to generate header
cJSON *commands = json_hdr_get(json, "commands", cJSON_Array, in_path);
cJSON *command = commands->child;
// Print this command
char keystr[8];
char *prevstr = NULL;
size_t i;
if(command) {
cJSON *key = json_hdr_get(command, "key", cJSON_String, in_path);
prevstr = key->valuestring;
}
// Print command entry for this command
strbuf_append_str(&hdrbuf, "##mccortex_");
strbuf_append_str(&hdrbuf, hex_rand_str(keystr, sizeof(keystr)));
strbuf_append_str(&hdrbuf, "=<prev=\"");
strbuf_append_str(&hdrbuf, prevstr ? prevstr : "NULL");
strbuf_append_str(&hdrbuf, "\",cmd=\"");
strbuf_append_str(&hdrbuf, cmd_get_cmdline());
strbuf_append_str(&hdrbuf, "\",cwd=\"");
strbuf_append_str(&hdrbuf, cmd_get_cwd());
strbuf_append_str(&hdrbuf, "\",version="CTX_VERSION">\n");
// Print previous commands
vcf_hdrtxt_append_commands(command, &hdrbuf, in_path);
// Print field definitions
if(is_breakpoint)
strbuf_append_str(&hdrbuf, "##INFO=<ID=BRKPNT,Number=1,Type=String,Description=\"Breakpoint call\">\n");
else
strbuf_append_str(&hdrbuf, "##INFO=<ID=BUBBLE,Number=1,Type=String,Description=\"Bubble call\">\n");
strbuf_sprintf(&hdrbuf, "##INFO=<ID=K%zu,Number=0,Type=Flag,Description=\"Found at k=%zu\">\n", kmer_size, kmer_size);
strbuf_append_str(&hdrbuf, "##FORMAT=<ID=GT,Number=1,Type=String,Description=\"Genotype\">\n");
strbuf_append_str(&hdrbuf, "##FILTER=<ID=PASS,Description=\"All filters passed\">\n");
// Print reference paths
strbuf_append_str(&hdrbuf, "##reference=");
strbuf_append_str(&hdrbuf, ref_paths[0]);
for(i = 1; i < nref_paths; i++) {
strbuf_append_char(&hdrbuf, ',');
strbuf_append_str(&hdrbuf, ref_paths[i]);
}
strbuf_append_str(&hdrbuf, "\n");
// Print contigs lengths
for(i = 0; i < nchroms; i++) {
strbuf_sprintf(&hdrbuf, "##contig=<ID=%s,length=%zu>\n",
chroms[i].name.b, chroms[i].seq.end);
}
// Print VCF column header
strbuf_append_str(&hdrbuf, "#CHROM\tPOS\tID\tREF\tALT\tQUAL\tFILTER\tINFO\tFORMAT");
if(is_breakpoint)
{
// Print a column for each sample
cJSON *graph_json = json_hdr_get(json, "graph", cJSON_Object, in_path);
cJSON *colours_json = json_hdr_get(graph_json, "colours", cJSON_Array, in_path);
cJSON *colour_json = colours_json->child;
if(colour_json == NULL) die("Missing colours");
for(; colour_json; colour_json = colour_json->next)
{
if(!json_hdr_colour_is_ref(colour_json)) {
cJSON *sample_json = json_hdr_get(colour_json, "sample", cJSON_String, in_path);
strbuf_append_str(&hdrbuf, "\t");
strbuf_append_str(&hdrbuf, sample_json->valuestring);
}
}
}
strbuf_append_char(&hdrbuf, '\n');
bcf_hdr_t *hdr = bcf_hdr_init("w");
if(bcf_hdr_parse(hdr, hdrbuf.b) != 0) die("Cannot construct VCF header");
strbuf_dealloc(&hdrbuf);
return hdr;
}
int ctx_join(int argc, char **argv)
{
struct MemArgs memargs = MEM_ARGS_INIT;
const char *out_path = NULL;
size_t use_ncols = 0;
GraphFileReader tmp_gfile;
GraphFileBuffer isec_gfiles_buf;
gfile_buf_alloc(&isec_gfiles_buf, 8);
// Arg parsing
char cmd[100], shortopts[100];
cmd_long_opts_to_short(longopts, shortopts, sizeof(shortopts));
int c;
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 '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 'N': cmd_check(!use_ncols, cmd); use_ncols = cmd_uint32_nonzero(cmd, optarg); break;
case 'i':
graph_file_reset(&tmp_gfile);
graph_file_open(&tmp_gfile, optarg);
if(file_filter_into_ncols(&tmp_gfile.fltr) > 1)
warn("Flattening intersection graph into colour 0: %s", optarg);
file_filter_flatten(&tmp_gfile.fltr, 0);
gfile_buf_push(&isec_gfiles_buf, &tmp_gfile, 1);
break;
case ':': /* BADARG */
case '?': /* BADCH getopt_long has already printed error */
// cmd_print_usage(NULL);
die("`"CMD" join -h` for help. Bad option: %s", argv[optind-1]);
default: abort();
}
}
GraphFileReader *igfiles = isec_gfiles_buf.b;
size_t num_igfiles = isec_gfiles_buf.len;
if(!out_path) cmd_print_usage("--out <out.ctx> required");
if(optind >= argc)
cmd_print_usage("Please specify at least one input graph file");
// optind .. argend-1 are graphs to load
size_t num_gfiles = (size_t)(argc - optind);
char **gfile_paths = argv + optind;
GraphFileReader *gfiles = ctx_calloc(num_gfiles, sizeof(GraphFileReader));
status("Probing %zu graph files and %zu intersect files", num_gfiles, num_igfiles);
// Check all binaries are valid binaries with matching kmer size
size_t i;
size_t ctx_max_cols = 0;
uint64_t min_intersect_num_kmers = 0, ctx_max_kmers = 0, ctx_sum_kmers = 0;
for(i = 0; i < num_gfiles; i++)
{
graph_file_open2(&gfiles[i], gfile_paths[i], "r", true, ctx_max_cols);
if(gfiles[0].hdr.kmer_size != gfiles[i].hdr.kmer_size) {
cmd_print_usage("Kmer sizes don't match [%u vs %u]",
gfiles[0].hdr.kmer_size, gfiles[i].hdr.kmer_size);
}
ctx_max_cols = MAX2(ctx_max_cols, file_filter_into_ncols(&gfiles[i].fltr));
ctx_max_kmers = MAX2(ctx_max_kmers, graph_file_nkmers(&gfiles[i]));
ctx_sum_kmers += graph_file_nkmers(&gfiles[i]);
}
// Probe intersection graph files
for(i = 0; i < num_igfiles; i++)
{
if(gfiles[0].hdr.kmer_size != igfiles[i].hdr.kmer_size) {
cmd_print_usage("Kmer sizes don't match [%u vs %u]",
gfiles[0].hdr.kmer_size, igfiles[i].hdr.kmer_size);
}
uint64_t nkmers = graph_file_nkmers(&igfiles[i]);
if(i == 0) min_intersect_num_kmers = nkmers;
else if(nkmers < min_intersect_num_kmers)
{
// Put smallest intersection binary first
SWAP(igfiles[i], igfiles[0]);
min_intersect_num_kmers = nkmers;
}
}
bool take_intersect = (num_igfiles > 0);
// If we are taking an intersection,
// all kmers intersection kmers will need to be loaded
if(take_intersect)
ctx_max_kmers = ctx_sum_kmers = min_intersect_num_kmers;
bool use_ncols_set = (use_ncols > 0);
bool output_to_stdout = (strcmp(out_path,"-") == 0);
// if(use_ncols == 0) use_ncols = 1;
if(use_ncols_set) {
if(use_ncols < ctx_max_cols && output_to_stdout)
die("I need %zu colours if outputting to STDOUT (--ncols)", ctx_max_cols);
if(use_ncols > ctx_max_cols) {
warn("I only need %zu colour%s ('--ncols %zu' ignored)",
ctx_max_cols, util_plural_str(ctx_max_cols), use_ncols);
use_ncols = ctx_max_cols;
}
}
else {
use_ncols = output_to_stdout ? ctx_max_cols : 1;
}
// Check out_path is writable
futil_create_output(out_path);
status("Output %zu cols; from %zu files; intersecting %zu graphs; ",
ctx_max_cols, num_gfiles, num_igfiles);
if(num_gfiles == 1 && num_igfiles == 0)
{
// Loading only one file with no intersection files
// Don't need to store a graph in memory, can filter as stream
// Don't actually store anything in the de Bruijn graph, but we need to
// pass it, so mock one up
dBGraph db_graph;
db_graph_alloc(&db_graph, gfiles[0].hdr.kmer_size,
file_filter_into_ncols(&gfiles[0].fltr), 0, 1024, 0);
graph_writer_stream_mkhdr(out_path, &gfiles[0], &db_graph, NULL, NULL);
graph_file_close(&gfiles[0]);
gfile_buf_dealloc(&isec_gfiles_buf);
ctx_free(gfiles);
db_graph_dealloc(&db_graph);
return EXIT_SUCCESS;
}
//
// Decide on memory
//
size_t bits_per_kmer, kmers_in_hash, graph_mem;
bits_per_kmer = sizeof(BinaryKmer)*8 +
(sizeof(Covg) + sizeof(Edges)) * 8 * use_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,
ctx_max_kmers, ctx_sum_kmers,
true, &graph_mem);
if(!use_ncols_set)
{
// Maximise use_ncols
size_t max_usencols = (memargs.mem_to_use*8) / bits_per_kmer;
use_ncols = MIN2(max_usencols, ctx_max_cols);
bits_per_kmer = sizeof(BinaryKmer)*8 +
(sizeof(Covg) + sizeof(Edges)) * 8 * use_ncols;
// Re-check memory used
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,
ctx_max_kmers, ctx_sum_kmers,
true, &graph_mem);
}
status("Using %zu colour%s in memory", use_ncols, util_plural_str(use_ncols));
cmd_check_mem_limit(memargs.mem_to_use, graph_mem);
// Create db_graph
dBGraph db_graph;
Edges *intersect_edges = NULL;
size_t edge_cols = (use_ncols + take_intersect);
db_graph_alloc(&db_graph, gfiles[0].hdr.kmer_size, use_ncols, use_ncols,
kmers_in_hash, DBG_ALLOC_COVGS);
// We allocate edges ourself since it's a special case
db_graph.col_edges = ctx_calloc(db_graph.ht.capacity*edge_cols, sizeof(Edges));
// Load intersection binaries
char *intsct_gname_ptr = NULL;
StrBuf intersect_gname;
strbuf_alloc(&intersect_gname, 1024);
if(take_intersect)
{
GraphLoadingPrefs gprefs = graph_loading_prefs(&db_graph);
gprefs.boolean_covgs = true; // covg++ only
for(i = 0; i < num_igfiles; i++)
{
graph_load(&igfiles[i], gprefs, NULL);
// Update intersect header
// note: intersection graphs all load exactly one colour into colour 0
graph_info_make_intersect(&igfiles[i].hdr.ginfo[0], &intersect_gname);
gprefs.must_exist_in_graph = true;
gprefs.must_exist_in_edges = db_graph.col_edges;
}
if(num_igfiles > 1)
{
// Remove nodes where covg != num_igfiles
HASH_ITERATE_SAFE(&db_graph.ht, remove_non_intersect_nodes,
db_graph.col_covgs, (Covg)num_igfiles, &db_graph.ht);
}
status("Loaded intersection set\n");
intsct_gname_ptr = intersect_gname.b;
for(i = 0; i < num_igfiles; i++) graph_file_close(&igfiles[i]);
// Reset graph info
for(i = 0; i < db_graph.num_of_cols; i++)
graph_info_init(&db_graph.ginfo[i]);
// Zero covgs
memset(db_graph.col_covgs, 0, db_graph.ht.capacity * sizeof(Covg));
// Use union edges we loaded to intersect new edges
intersect_edges = db_graph.col_edges;
db_graph.col_edges += db_graph.ht.capacity;
}
bool kmers_loaded = take_intersect, colours_loaded = false;
graph_writer_merge_mkhdr(out_path, gfiles, num_gfiles,
kmers_loaded, colours_loaded, intersect_edges,
intsct_gname_ptr, &db_graph);
if(take_intersect)
db_graph.col_edges -= db_graph.ht.capacity;
for(i = 0; i < num_gfiles; i++) graph_file_close(&gfiles[i]);
strbuf_dealloc(&intersect_gname);
gfile_buf_dealloc(&isec_gfiles_buf);
ctx_free(gfiles);
db_graph_dealloc(&db_graph);
return EXIT_SUCCESS;
}
int ctx_links(int argc, char **argv)
{
size_t limit = 0;
const char *link_out_path = NULL, *csv_out_path = NULL, *plot_out_path = NULL;
const char *thresh_path = NULL, *hist_path = NULL;
size_t hist_distsize = 0, hist_covgsize = 0;
size_t cutoff = 0;
bool clean = false;
// Arg parsing
char cmd[100];
char shortopts[300];
cmd_long_opts_to_short(longopts, shortopts, sizeof(shortopts));
int c;
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(!link_out_path, cmd); link_out_path = optarg; break;
case 'f': cmd_check(!futil_get_force(), cmd); futil_set_force(true); break;
case 'l': cmd_check(!csv_out_path, cmd); csv_out_path = optarg; break;
case 'c': cmd_check(!cutoff, cmd); cutoff = cmd_size(cmd, optarg); clean = true; break;
case 'L': cmd_check(!limit, cmd); limit = cmd_size(cmd, optarg); break;
case 'P': cmd_check(!plot_out_path, cmd); plot_out_path = optarg; break;
case 'T': cmd_check(!thresh_path, cmd); thresh_path = optarg; break;
case 'H': cmd_check(!hist_path, cmd); hist_path = optarg; break;
case 'C': cmd_check(!hist_covgsize, cmd); hist_covgsize = cmd_size(cmd, optarg); break;
case 'D': cmd_check(!hist_distsize, cmd); hist_distsize = cmd_size(cmd, optarg); break;
case ':': /* BADARG */
case '?': /* BADCH getopt_long has already printed error */
// cmd_print_usage(NULL);
die("`"CMD" links -h` for help. Bad option: %s", argv[optind-1]);
default: ctx_assert2(0, "shouldn't reach here: %c", c);
}
}
if(hist_distsize && !hist_path) cmd_print_usage("--max-dist without --covg-hist");
if(hist_covgsize && !hist_path) cmd_print_usage("--max-covg without --covg-hist");
// Defaults
if(!hist_distsize) hist_distsize = DEFAULT_MAX_DIST;
if(!hist_covgsize) hist_covgsize = DEFAULT_MAX_COVG;
if(optind + 1 != argc) cmd_print_usage("Wrong number of arguments");
const char *ctp_path = argv[optind];
bool list = (csv_out_path != NULL);
bool plot = (plot_out_path != NULL);
bool save = (link_out_path != NULL);
bool hist_covg = (thresh_path != NULL || hist_path != NULL);
size_t plot_kmer_idx = (limit == 0 ? 0 : limit - 1);
if(clean && !save)
cmd_print_usage("Need to give --out <out.ctp.gz> with --clean");
if(!save && !list && !plot && !hist_covg)
cmd_print_usage("Please specify one of --plot, --list or --clean");
if(link_out_path && hist_covg && strcmp(link_out_path,"-") == 0)
cmd_print_usage("Outputing both cleaning threshold (-T) and links (-o) to STDOUT!");
// Open input file
FILE *list_fh = NULL, *plot_fh = NULL, *link_tmp_fh = NULL;
FILE *thresh_fh = NULL, *hist_fh = NULL;
gzFile link_gz = NULL;
// Check file don't exist or that we can overwrite
// Will ignore if path is null
bool err = false;
err |= futil_check_outfile(csv_out_path);
err |= futil_check_outfile(plot_out_path);
err |= futil_check_outfile(link_out_path);
err |= futil_check_outfile(thresh_path);
err |= futil_check_outfile(hist_path);
if(err) die("Use -f,--force to overwrite files");
StrBuf link_tmp_path;
strbuf_alloc(&link_tmp_path, 1024);
GPathReader ctpin;
memset(&ctpin, 0, sizeof(ctpin));
gpath_reader_open(&ctpin, ctp_path);
size_t ncols = file_filter_into_ncols(&ctpin.fltr);
size_t kmer_size = gpath_reader_get_kmer_size(&ctpin);
cJSON *newhdr = cJSON_Duplicate(ctpin.json, 1);
if(ncols != 1) die("Can only clean a single colour at a time. Sorry.");
uint64_t (*hists)[hist_covgsize] = NULL;
if(hist_covg) {
hists = ctx_calloc(hist_distsize, sizeof(hists[0]));
}
if(hist_path && (hist_fh = futil_fopen_create(hist_path, "w")) == NULL)
die("Cannot open file: %s", hist_path);
if(thresh_path && (thresh_fh = futil_fopen_create(thresh_path, "w")) == NULL)
die("Cannot open file: %s", thresh_path);
if(limit)
status("Limiting to the first %zu kmers", limit);
if(clean)
{
timestamp();
message(" Cleaning coverage below %zu", cutoff);
message("\n");
}
if(save)
{
// Check we can find the fields we need
cJSON *links_json = json_hdr_get(newhdr, "paths", cJSON_Object, link_out_path);
cJSON *nkmers_json = json_hdr_get(links_json, "num_kmers_with_paths", cJSON_Number, link_out_path);
cJSON *nlinks_json = json_hdr_get(links_json, "num_paths", cJSON_Number, link_out_path);
cJSON *nbytes_json = json_hdr_get(links_json, "path_bytes", cJSON_Number, link_out_path);
if(!nkmers_json || !nlinks_json || !nbytes_json)
die("Cannot find required header entries");
// Create a random temporary file
link_tmp_fh = create_tmp_file(&link_tmp_path, link_out_path);
status("Saving output to: %s", link_out_path);
status("Temporary output: %s", link_tmp_path.b);
// Open output file
if((link_gz = futil_gzopen_create(link_out_path, "w")) == NULL)
die("Cannot open output link file: %s", link_out_path);
// Need to open output file first so we can get absolute path
// Update the header to include this command
json_hdr_add_curr_cmd(newhdr, link_out_path);
}
if(list)
{
status("Listing to %s", csv_out_path);
if((list_fh = futil_fopen_create(csv_out_path, "w")) == NULL)
die("Cannot open output CSV file %s", csv_out_path);
// Print csv header
fprintf(list_fh, "SeqLen,Covg\n");
}
if(plot)
{
status("Plotting kmer %zu to %s", plot_kmer_idx, plot_out_path);
if((plot_fh = futil_fopen_create(plot_out_path, "w")) == NULL)
die("Cannot open output .dot file %s", plot_out_path);
}
SizeBuffer countbuf, jposbuf;
size_buf_alloc(&countbuf, 16);
size_buf_alloc(&jposbuf, 1024);
StrBuf kmerbuf, juncsbuf, seqbuf, outbuf;
strbuf_alloc(&kmerbuf, 1024);
strbuf_alloc(&juncsbuf, 1024);
strbuf_alloc(&seqbuf, 1024);
strbuf_alloc(&outbuf, 1024);
bool link_fw;
size_t njuncs;
size_t knum, nlinks, num_links_exp = 0;
LinkTree ltree;
ltree_alloc(<ree, kmer_size);
LinkTreeStats tree_stats;
memset(&tree_stats, 0, sizeof(tree_stats));
size_t init_num_links = 0, num_links = 0;
for(knum = 0; !limit || knum < limit; knum++)
{
ltree_reset(<ree);
if(!gpath_reader_read_kmer(&ctpin, &kmerbuf, &num_links_exp)) break;
ctx_assert2(kmerbuf.end == kmer_size, "Kmer incorrect length %zu != %zu",
kmerbuf.end, kmer_size);
// status("kmer: %s", kmerbuf.b);
for(nlinks = 0;
gpath_reader_read_link(&ctpin, &link_fw, &njuncs,
&countbuf, &juncsbuf,
&seqbuf, &jposbuf);
nlinks++)
{
ltree_add(<ree, link_fw, countbuf.b[0], jposbuf.b,
juncsbuf.b, seqbuf.b);
}
if(nlinks != num_links_exp)
warn("Links count mismatch %zu != %zu", nlinks, num_links_exp);
if(hist_covg)
{
ltree_update_covg_hists(<ree, (uint64_t*)hists,
hist_distsize, hist_covgsize);
}
if(clean)
{
ltree_clean(<ree, cutoff);
}
// Accumulate statistics
ltree_get_stats(<ree, &tree_stats);
num_links = tree_stats.num_links - init_num_links;
init_num_links = tree_stats.num_links;
if(list)
{
ltree_write_list(<ree, &outbuf);
if(fwrite(outbuf.b, 1, outbuf.end, list_fh) != outbuf.end)
die("Cannot write CSV file to: %s", csv_out_path);
strbuf_reset(&outbuf);
}
if(save && num_links)
{
ltree_write_ctp(<ree, kmerbuf.b, num_links, &outbuf);
if(fwrite(outbuf.b, 1, outbuf.end, link_tmp_fh) != outbuf.end)
die("Cannot write ctp file to: %s", link_tmp_path.b);
strbuf_reset(&outbuf);
}
if(plot && knum == plot_kmer_idx)
{
status("Plotting tree...");
ltree_write_dot(<ree, &outbuf);
if(fwrite(outbuf.b, 1, outbuf.end, plot_fh) != outbuf.end)
die("Cannot write plot DOT file to: %s", plot_out_path);
strbuf_reset(&outbuf);
}
}
gpath_reader_close(&ctpin);
cJSON *links_json = json_hdr_get(newhdr, "paths", cJSON_Object, link_out_path);
cJSON *nkmers_json = json_hdr_get(links_json, "num_kmers_with_paths", cJSON_Number, link_out_path);
cJSON *nlinks_json = json_hdr_get(links_json, "num_paths", cJSON_Number, link_out_path);
cJSON *nbytes_json = json_hdr_get(links_json, "path_bytes", cJSON_Number, link_out_path);
status("Number of kmers with links %li -> %zu", nkmers_json->valueint, tree_stats.num_trees_with_links);
status("Number of links %li -> %zu", nlinks_json->valueint, tree_stats.num_links);
status("Number of bytes %li -> %zu", nbytes_json->valueint, tree_stats.num_link_bytes);
if(save)
{
// Update JSON
nkmers_json->valuedouble = nkmers_json->valueint = tree_stats.num_trees_with_links;
nlinks_json->valuedouble = nlinks_json->valueint = tree_stats.num_links;
nbytes_json->valuedouble = nbytes_json->valueint = tree_stats.num_link_bytes;
char *json_str = cJSON_Print(newhdr);
if(gzputs(link_gz, json_str) != (int)strlen(json_str))
die("Cannot write ctp file to: %s", link_out_path);
free(json_str);
gzputs(link_gz, "\n\n");
gzputs(link_gz, ctp_explanation_comment);
gzputs(link_gz, "\n");
fseek(link_tmp_fh, 0, SEEK_SET);
char *tmp = ctx_malloc(4*ONE_MEGABYTE);
size_t s;
while((s = fread(tmp, 1, 4*ONE_MEGABYTE, link_tmp_fh)) > 0) {
if(gzwrite(link_gz, tmp, s) != (int)s)
die("Cannot write to output: %s", link_out_path);
}
ctx_free(tmp);
gzclose(link_gz);
fclose(link_tmp_fh);
}
// Write histogram to file
if(hist_fh)
{
size_t i, j;
fprintf(hist_fh, " ");
for(j = 1; j < hist_covgsize; j++) fprintf(hist_fh, ",covg.%02zu", j);
fprintf(hist_fh, "\n");
for(i = 1; i < hist_distsize; i++) {
fprintf(hist_fh, "dist.%02zu", i);
for(j = 1; j < hist_covgsize; j++) {
fprintf(hist_fh, ",%"PRIu64, hists[i][j]);
}
fprintf(hist_fh, "\n");
}
}
if(thresh_fh)
{
// Use median of first five cutoffs
print_suggest_cutoff(6, hist_covgsize, hists, thresh_fh);
}
if(hist_fh && hist_fh != stdout) fclose(hist_fh);
if(list)
{
fclose(list_fh);
}
if(plot)
{
fclose(plot_fh);
}
ctx_free(hists);
cJSON_Delete(newhdr);
strbuf_dealloc(&link_tmp_path);
ltree_dealloc(<ree);
size_buf_dealloc(&countbuf);
size_buf_dealloc(&jposbuf);
strbuf_dealloc(&kmerbuf);
strbuf_dealloc(&juncsbuf);
strbuf_dealloc(&seqbuf);
strbuf_dealloc(&outbuf);
return EXIT_SUCCESS;
}
