1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421
|
#include <stdio.h>
#include <stdlib.h>
#include <time.h>
#include <string.h>
#include <unistd.h>
#include <sys/types.h>
#include <sys/stat.h> // for mkdir
#include <inttypes.h>
#include <stdint.h>
#include <algorithm> // for max/min
#include <vector> // for sorting_kmers
#include <sys/time.h>
#define NNKS 4 // default minimal abundance for solidity
int max_memory; // the most memory one should alloc at any time, in MB
int order=0; // in kissnp, don't change it, it should be 0
#include "../minia/Bank.h"
#include "../minia/Hash16.h"
#include "../minia/Set.h"
#include "../minia/Pool.h"
#include "../minia/Bloom.h"
#include "../minia/Debloom.h"
#include "../minia/Utils.h"
#include "../minia/SortingCount.h"
#include "../minia/Terminator.h"
#include "../minia/Kmer.h"
#include "../minia/rvalues.h"
//#include "SNP.h"
//#include "Kmer_for_kissnp2.h"
#include "Starter_Bank.h"
#include "commons.h"
extern int size_seeds;
extern string prefix_trashable;
using namespace std;
/////////////////////// FOR MINIA //////////////////
extern int sizeKmer;
int64_t genome_size;
Bloom * bloo1;
int threshold;
BinaryBank * SolidKmers;
BranchingTerminator * terminator;
extern unsigned int nbits_nbseeds;
extern uint64_t mask_nbseed ;
extern uint64_t mask_offset_seed;
////////////////////////////////////////////////////
char * getVersion(){
return (char *)"2.0.5.deb - Copyright INRIA - CeCILL License";
}
void print_usage_and_exit(char * name){
fprintf (stderr, "NAME\nmapsembler, version %s\n", getVersion());
fprintf (stderr, "\nSYNOPSIS\n%s <starters.fasta> <readsC1.fasta> [<readsC2.fasta> [<readsC3.fasta] ...] [-E] [-t extension_type] [-q value] [-k value] [-c value] [-d authorized_distance] [-e error_threshold] [-g value] [-i index_name] [-o name] [-m name] [-h]\n", name);
fprintf (stderr, "\nDESCRIPTION\n");
fprintf (stderr, "\t Mapsembler2 is a targeted assembly software. It takes as input a set of NGS raw reads (fasta or fastq, gzipped or not) and a set of input sequences (starters). It first determines if each starter is read-coherent, e.g. whether reads confirm the presence of each starter in the original sequence. Then for each read-coherent starter, Mapsembler2 outputs its sequence neighborhood as a linear sequence or as a graph, depending on the user choice.\n");
fprintf (stderr, "\nOPTIONS\n");
//fprintf (stderr, "\t -c Write the coverage of each position of the sequences. The fasta file will have 3 lines (instead of one) starting with \'>\' before the sequences themselves\n");
fprintf (stderr, "\t -E Extend only: avoid the mapping+substarter generation phase\n");
fprintf (stderr, "\t -t extension_type. Default: 1\n");
fprintf (stderr, "\t 1: a strict sequence: any branching stops the extension\n");
fprintf (stderr, "\t 2: a consensus sequence: contiging approach\n");
fprintf (stderr, "\t 3: a strict graph: any branching is conserved in the graph\n");
fprintf (stderr, "\t 4: a consensus graph: \"small\" polymorphism is merged, but \"large\" structures are represented\n");
fprintf (stderr, "\t -q size_seed: will use seeds of length size_seed during the mapping process. Default: 25, value should be in [5-31]\n");
fprintf (stderr, "\t -k size_kmers: Size of the k-mers used duriung the extension phase Default: 31. Accepted range, depends on the compilation (make k=42 for instance) \n");
fprintf (stderr, "\t -c min_coverage: a sequence is covered by at least min_coverage coherent reads. Default: 2\n");
fprintf (stderr, "\t -d authorized_distance: a substarter is distant by at most authorized_distance substitutions. Default: 2\n");
fprintf (stderr, "\t -e error_threshold: a nucleotide is corrected if occurs less than error_threshold times. Default: 2\n");
fprintf (stderr, "\t -g estimated_genome_size: estimation of the size of the genome whose reads come from. \n \t It is in bp, does not need to be accurate, only controls memory usage. Default: 3 billion\n");
fprintf (stderr, "\t -n node_len: limit max of nodes length. Default: 1000000\n");
fprintf (stderr, "\t -l graph_max_depth: limit max of graph depth.Default: 10000\n");
fprintf (stderr, "\t -i index_name: stores the index files in files starting with this prefix name. Can be re-used latter. Default: \"index\"\n");
fprintf (stderr, "\t IF THE FILE \"index_name.bloom\" EXISTS: the index is not re-created \n");
fprintf (stderr, "\t -o file_name_prefix: where to write outputs. Default: \"res_mapsembler\" \n");
fprintf (stderr, "\t -p search_mod: kind of prosses LARGEUR or PROFONDEUR. Default: LARGEUR \n");
fprintf (stderr, "\t -m file_name: write in file \"file_name\" the reads mapped on starters\n");
fprintf (stderr, "\t -h prints this message and exit\n");
exit(0);
}
int main(int argc, char *argv[])
{
setvbuf(stdout,(char*)NULL,_IONBF,0); // disable buffering for printf()
size_seeds=25;
int min_coverage=2; // minimal number of reads per positions extending the initial fragment
int error_threshold=2; //comment todo
int authorized_distance=2; // comment todo et comrendre diff
char extension_type=1; // kind of extensions
int max_graph_depth = 10000; //detph graph limit
int max_nodes = 1000000; //node length limit
int process_type = 1; //kind of process : 1 == LARGEUR, 2 == PROFONDEUR
parcours_t search_mode = LARGEUR;
bool export_mapped_reads=false;
char * file_read_mapped;
bool make_mapping = true;
genome_size=3000000000;
if(argc<2){
print_usage_and_exit(argv[0]);
}
char * toCheck_file =strdup(argv[1]);
FILE * out_results = stdout;
char * index_file_prefix = new char [4096];
char * res_prefix = new char [4096];
strcpy(index_file_prefix,"index");
strcpy(res_prefix,"res_mapsembler");
////////////////////////////////// GETTING THE OPTIONS /////////////////////////////////////
sizeKmer=31;
// GET ALL THE READ FILES
// find the number of read sets
int number_of_read_sets=0;
while(number_of_read_sets+2<argc && argv[number_of_read_sets+2][0]!='-') number_of_read_sets++;
char ** reads_file_names = (char **) malloc(sizeof(char *)*number_of_read_sets);
// copy the read set file names
number_of_read_sets=0;
while(number_of_read_sets+2<argc && argv[number_of_read_sets+2][0]!='-'){
reads_file_names[number_of_read_sets]=strdup(argv[number_of_read_sets+2]);
number_of_read_sets++;
}
while (1)
{
int temoin = getopt (argc-number_of_read_sets-1, &argv[number_of_read_sets+1], "t:c:e:d:x:y:o:f:k:q:g:i:m:h:E");
if (temoin == -1){
break;
}
switch (temoin)
{
case 'E':
make_mapping=false;
printf(" Mapsembler will extend the starters, not checking their read-coherency nor generating substarters\n");
break;
case 't':
extension_type=atoi(optarg);
printf(" Extension_Type=%d\n", extension_type);
break;
case 'c':
min_coverage=atoi(optarg);
printf(" Will consider as read coherent if coverage is at least %d\n", min_coverage);
break;
case 'e':
error_threshold=atoi(optarg);
printf(" Will use error_threshold=%d\n", error_threshold);
break;
case 'd':
authorized_distance=atoi(optarg);
printf(" Will use authorized_distance=%d\n", authorized_distance);
break;
case 'g':
genome_size=atoll(optarg);
printf(" Will use genome_size=%llu\n", genome_size); //TODO format
break;
case 'x':
max_nodes=atoi(optarg);
printf(" Will use node_len=%d\n", max_nodes); //TODO format
break;
case 'y':
max_graph_depth=atoi(optarg);
printf(" Will use max_graph_depth=%d\n", max_graph_depth); //TODO format
break;
case 'o':
strcpy(res_prefix,optarg);
printf(" Will output results in file prefixed with %s\n", optarg);
break;
case 'f':
process_type=atoi(optarg);
if(process_type == 1){
search_mode = LARGEUR;
}else if(process_type == 2){
search_mode = PROFONDEUR;
}
printf(" Will use process_type %d\n", process_type);
break;
case 'm':
file_read_mapped=strdup(optarg);
export_mapped_reads=true;
printf(" Will output mapped reads in file: %s\n", file_read_mapped);
break;
case 'i':
strcpy(index_file_prefix, optarg);
printf(" Will store read index in files prefixed with: %s\n", index_file_prefix);
break;
case 'q':
size_seeds=atoi(optarg);
if(size_seeds<5){
fprintf(stderr,"%d too small to be used as seed length. kisSnpCheckReads continues with seeds of length 5\n",size_seeds);
size_seeds=5;
}
if(size_seeds>31){
fprintf(stderr,"%d too big to be used as seed length. kisSnpCheckReads continues with seeds of length 31\n",size_seeds);
size_seeds=31;
}
printf(" Will use seeds of length %d for mapping\n", size_seeds);
break;
case 'k':
sizeKmer=atoi(optarg);
// let's make k even for now, because i havnt thought of how to handle palindromes (dont want to stop on them)
if (sizeKmer%2==0) //TODO: verif comptaibilité minia
{
sizeKmer-=1;
printf("Need odd kmer size to avoid palindromes. I've set kmer size to %d.\n",sizeKmer);
}
if (sizeKmer>(sizeof(kmer_type)*4))
{
printf("Max kmer size on this compiled version is %d\n",(int)sizeof(kmer_type)*4);
exit(1);
}
printf(" Will use kmers of length %d for extensions\n", sizeKmer);
break;
case 'h':
print_usage_and_exit(argv[0]);
// case 'v':
// printf(" VERBOSE mode\n");
// verbose=1;
// break;
case '-':
if(strcmp("version", optarg)==0){
printf("kissReads version %s\n", getVersion());
exit(0);
}
printf(" what next ? %s\n",optarg);
break;
default:
// printf ("Unknown option %c\n", temoin);
print_usage_and_exit(argv[0]);
}
}
if ( argc - optind <2)
{
print_usage_and_exit(argv[0]);
}
if(!make_mapping && export_mapped_reads){
fprintf(stderr,"Warning options -E and -m are incompatible. I won't output mapped reads (disabling -m option)");
export_mapped_reads=false;
}
sprintf(prefix,"%s_k_%d_c_%d", index_file_prefix, sizeKmer, min_coverage);
char final_res_prefix [4096];
sprintf(final_res_prefix,"%s_k_%d_q_%d_c_%d_t_%d", res_prefix, sizeKmer, size_seeds, min_coverage, (int)extension_type);
init_static_variables();
Bank *reads = new Bank(reads_file_names,number_of_read_sets);
Starter_Bank *starter_bank;
Extension_Bank * extensions;
///// USED BOTH FOR mapping and extensions
nbits_nbseeds = 8*sizeof(uint64_t)- NBITS_OFFSET_SEED ;
mask_nbseed = ( 1ULL << (uint64_t) nbits_nbseeds ) -1 ;
mask_offset_seed = (1ULL << (NBITS_OFFSET_SEED)) -1 ;
////////////////////////////////// READ COHERENCE /////////////////////////////////////////
if(make_mapping){
printf("indexing starters\n");
Fragment_Bank *fragment_bank= new Fragment_Bank(); // loading the fragment sequences
fragment_bank->load_fragments(argv[1]);
fragment_bank->index_fragments_and_init_vector();
printf("map reads, checking the read coherency and discovering subfragments\n");
fragment_bank->map_reads(reads, authorized_distance);
// fragment_bank->show_alignments();
if(export_mapped_reads) fragment_bank->output_mapped_reads(file_read_mapped);
starter_bank = new Starter_Bank(fragment_bank);
delete(fragment_bank);
printf("generate substarters\n");
extensions = starter_bank->generate_all_substarters(min_coverage, authorized_distance, error_threshold, sizeKmer);
if(extensions->all_extensions.size()==0){
printf("No starter was read coherent, we stop here\n");
exit(0);
}
}
else{
Fragment_Bank *fragment_bank= new Fragment_Bank(); // loading the fragment sequences
fragment_bank->load_fragments(argv[1]);
starter_bank = new Starter_Bank(fragment_bank);
extensions = starter_bank->generate_all_substarter_nomapping(sizeKmer);
}
////////////////////////////////// EXTENSIONS //////////////////////////////////////////
extensions->set_extension_type(extension_type);
printf("Indexing reads, using minia approach, generating file prefixed with \"%s\"\n", prefix);
kmerMask=(((kmer_type)1)<<(sizeKmer*2))-1;
double lg2 = log(2);
NBITS_PER_KMER = rvalues[sizeKmer][1];
//NBITS_PER_KMER = log(16*sizeKmer*(lg2*lg2))/(lg2*lg2); // needed to process argv[5]
// solidity
nks =NNKS;
nks = min_coverage;
if (nks<=0) nks=1; // min abundance can't be 0
int estimated_BL1 = max( (int)ceilf(log2f(genome_size * NBITS_PER_KMER )), 1);
uint64_t estimated_nb_FP = (uint64_t)(genome_size * 4 * powf(0.6,11)); // just indicative
max_memory = max( (1LL << estimated_BL1)/8LL /1024LL/1024LL, 1LL );
// printf("estimated values: nbits Bloom %i, nb FP %lld, max memory %i MB\n",estimated_BL1,estimated_nb_FP,max_memory);
// shortcuts to go directly to assembly using serialized bloom and serialized hash
int START_FROM_SOLID_KMERS=0; // if = 0, construct the fasta file of solid kmers, if = 1, start directly from that file
int LOAD_FALSE_POSITIVE_KMERS=0; // if = 0, construct the fasta file of false positive kmers (debloom), if = 1, load that file into the hashtable
int NO_FALSE_POSITIVES_AT_ALL=0; // if = 0, normal behavior, if = 1, don't load false positives (will be a probabilistic de bruijn graph)
//tester si le fichier prefix.bloom exist et faire:
if( access( (string(prefix)+string(".debloom")).c_str(), F_OK ) != -1 &&
access( (string(prefix)+string(".debloom2")).c_str(), F_OK ) != -1 &&
access( (string(prefix)+string(".false_positive_kmers")).c_str(), F_OK ) != -1 &&
access( (string(prefix)+string(".solid_kmers_binary")).c_str(), F_OK ) != -1){
printf("THE INDEX %s.... were already created we use them\n", prefix);
START_FROM_SOLID_KMERS=1;
LOAD_FALSE_POSITIVE_KMERS=1;
}
else
printf("CREATING THE index %s.... \n", prefix);
// fprintf (stderr,"taille cell %lu \n", sizeof(cell<kmer_type>));
STARTWALL(0);
// counter kmers, write solid kmers to disk, insert them into bloo1
if (!START_FROM_SOLID_KMERS)
{
int max_disk_space = 0; // let dsk decide
int verbose = 0;
bool write_count = false;
sorting_count(reads,prefix, max_memory, max_disk_space, write_count, verbose);
}
// debloom, write false positives to disk, insert them into false_positives
if (! LOAD_FALSE_POSITIVE_KMERS)
{
debloom(order, max_memory);
}
bloo1 = bloom_create_bloo1((BloomCpt *)NULL);
// load false positives from disk into false_positives
false_positives = load_false_positives_cascading4();
// load branching kmers
//BinaryBank *SolidKmers = new BinaryBank(return_file_name(solid_kmers_file),sizeof(kmer_type),0);
int LOAD_BRANCHING_KMERS=0;
if (LOAD_BRANCHING_KMERS)
{
BinaryBank *BranchingKmers = new BinaryBank(return_file_name(branching_kmers_file),sizeof(kmer_type),false);
terminator = new BranchingTerminator(BranchingKmers,SolidKmers, bloo1,false_positives);
BranchingKmers->close();
delete BranchingKmers;
}
else{
BinaryBank *SolidKmers = new BinaryBank(return_file_name(solid_kmers_file),sizeof(kmer_type),0);
terminator = new BranchingTerminator(SolidKmers,genome_size, bloo1,false_positives);
SolidKmers->close();
delete SolidKmers;
}
printf("Generating %d extensions\n", (int) extensions->all_extensions.size());
extensions->perform_extensions(prefix, max_graph_depth, max_nodes, search_mode);
printf("Format results\n");
starter_bank->format_results(extension_type, extensions, final_res_prefix);
// CLEANING THE TEMP FILES
string command;
// command = string("rm -f ")+prefix_trashable.c_str()+string("*"); system((char *)command.c_str());
//command = string("rm -f *.json_edges"); system((char *)command.c_str());
//command = string("rm -f *.json_nodes"); system((char *)command.c_str());
//command = string("rm -f *.json_substarters"); system((char *)command.c_str());
STOPWALL(0,"Total");
delete bloo1;
delete terminator;
delete starter_bank;
delete extensions;
delete reads;
free(toCheck_file);
for(int i=0;i<number_of_read_sets;i++) free(reads_file_names[i]); free(reads_file_names);
printf("all done, results are in %s.%s\n", final_res_prefix, extension_type<3?"fasta":"json");
return 0;
}
|