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 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804 805 806 807 808 809 810 811 812 813 814 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843 844 845 846 847 848 849 850 851 852 853 854 855 856 857 858 859 860 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880 881 882 883 884 885 886 887 888 889
|
package hiseq;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.concurrent.locks.ReadWriteLock;
import java.util.concurrent.locks.ReentrantReadWriteLock;
import aligner.MicroAligner3;
import aligner.MicroIndex3;
import barcode.BarcodeStats;
import bloom.BloomFilter;
import bloom.BloomFilterCorrector;
import bloom.KmerCountAbstract;
import dna.AminoAcid;
import dna.Data;
import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import jgi.Dedupe;
import kmer.AbstractKmerTable;
import kmer.HashArray1D;
import kmer.ScheduleMaker;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import structures.IntList;
import structures.ListNum;
import structures.LongList;
import template.Accumulator;
import template.ThreadWaiter;
//Example command: plotflowcell.sh in=lane1.fq.gz size=999999 dump=dump.txt expected=expected.txt bloom multithreaded -Xmx40g
/**
* Analyzes a flow cell for low-quality areas.
* Removes reads in the low-quality areas.
*
* @author Brian Bushnell
* @date August 31, 2016
*
*/
public class PlotFlowCell implements Accumulator<PlotFlowCell.WorkerThread> {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
* @param args Command line arguments
*/
public static void main(String[] args){
Timer t=new Timer();
PlotFlowCell x=new PlotFlowCell(args);
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
/**
* Constructor.
* @param args Command line arguments
*/
public PlotFlowCell(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
//Set shared static variables
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Shared.threads());
//Create a parser object
Parser parser=new Parser();
boolean setInterleaved=false; //Whether interleaved was explicitly set.
//Parse each argument
for(int i=0; i<args.length; i++){
String arg=args[i];
//Break arguments into their constituent parts, in the form of "a=b"
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
}else if(a.equals("divisor") || a.equals("size")){
Tile.xSize=Tile.ySize=Parse.parseIntKMG(b);
}else if(a.equals("xdivisor") || a.equals("xsize")){
Tile.xSize=Parse.parseIntKMG(b);
}else if(a.equals("ydivisor") || a.equals("ysize")){
Tile.ySize=Parse.parseIntKMG(b);
}else if(a.equals("target")){
targetAverageReads=Parse.parseIntKMG(b);
}else if(a.equals("dump") || a.equals("out")){
dump=b;
}else if(a.equals("indump") || a.equals("ind") || a.equals("dumpin")){
dumpIn=b;
}else if(a.equals("pound")){
pound=Parse.parseBoolean(b);
}else if(a.equals("loadkmers") || a.equals("usekmers")){
loadKmers=Parse.parseBoolean(b);
}else if(a.equals("allkmers")){
kmersPerRead=(Parse.parseBoolean(b) ? 0 : 1);
}else if(a.equals("kmersperread")){
kmersPerRead=Integer.parseInt(b);
}else if(a.equals("multithreaded")){
multithreadedLoad=multithreadedFill=Parse.parseBoolean(b);
}else if(a.equals("multiload")){
multithreadedLoad=Parse.parseBoolean(b);
}else if(a.equals("multifill")){
multithreadedFill=Parse.parseBoolean(b);
}else if(a.equals("bloom")){
bloomMode=Parse.parseBoolean(b);
}else if(a.equals("bits") || a.equals("cbits")){
cbits=Integer.parseInt(b);
}else if(a.equals("hashes")){
hashes=Integer.parseInt(b);
}else if(a.equals("ref")){
refPath=b;
}else if(a.equals("minid")){
minIdentity=Float.parseFloat(b);
if(minIdentity>1) {minIdentity/=100;}
}else if(a.equals("kalign") || a.equals("alignk")){
alignK=Integer.parseInt(b);
}else if(a.equals("expectedbarcodes") || a.equals("expected") || a.equals("barcodes")){
expectedBarcodes=b;
}else if(a.equals("shortheader")){
MicroTile.shortHeader=Parse.parseBoolean(b);
}else if(a.equals("longheader")){
MicroTile.shortHeader=!Parse.parseBoolean(b);
}
else if(a.equals("minpolyg")){
MicroTile.MIN_POLY_G=Integer.parseInt(b);
}else if(a.equals("trackcycles")){
MicroTile.TRACK_CYCLES=Parse.parseBoolean(b);
}else if(a.equals("extra")){
if(b!=null) {
for(String s : b.split(",")) {
extra.add(s);
}
}
}
else if(a.equals("parse_flag_goes_here")){
//Set a variable here
}else if(parser.parse(arg, a, b)){//Parse standard flags in the parser
//do nothing
}else if(b==null && new File(arg).exists()){
extra.add(b);
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
// throw new RuntimeException("Unknown parameter "+args[i]);
}
}
{//Process parser fields
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=parser.overwrite;
append=parser.append;
setInterleaved=parser.setInterleaved;
in1=parser.in1;
in2=parser.in2;
if(dump==null && parser.out1!=null){dump=parser.out1;}
extin=parser.extin;
}
if("phix".equalsIgnoreCase(refPath)) {
refPath=Data.findPath("?phix2.fa.gz");
}
//Do input file # replacement
if(in1!=null && in2==null && in1.indexOf('#')>-1 && !new File(in1).exists()){
in2=in1.replace("#", "2");
in1=in1.replace("#", "1");
}
//Adjust interleaved detection based on the number of input files
if(in2!=null){
if(FASTQ.FORCE_INTERLEAVED){outstream.println("Reset INTERLEAVED to false because paired input files were specified.");}
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=false;
}
assert(FastaReadInputStream.settingsOK());
//Ensure there is an input file
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
//Adjust the number of threads for input file reading
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
//Adjust interleaved settings based on number of output files
if(!setInterleaved){
assert(in1!=null) : "\nin1="+in1+"\nin2="+in2+"\n";
if(in2!=null){ //If there are 2 input streams.
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=false;
outstream.println("Set INTERLEAVED to "+FASTQ.FORCE_INTERLEAVED);
}
}
//Ensure output files can be written
if(!Tools.testOutputFiles(overwrite, append, false, dump)){
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output file "+dump+"\n");
}
//Ensure input files can be read
if(!Tools.testInputFiles(false, true, in1, in2)){
throw new RuntimeException("\nCan't read some input files.\n");
}
//Ensure that no file was specified multiple times
if(!Tools.testForDuplicateFiles(true, in1, in2, dump)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
//Create input FileFormat objects
ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
ffin2=FileFormat.testInput(in2, FileFormat.FASTQ, extin, true, true);
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
public void process(Timer t){
final int oldThreads=Shared.threads();
Shared.capThreads(kmersPerRead>0 ? 64 : 64);
barcodeStats=loadBarcodes(expectedBarcodes);
//Reset counters
readsProcessed=0;
basesProcessed=0;
MicroIndex3 index=(refPath==null ? null : new MicroIndex3(alignK, 0, refPath, false));
mapper=(index==null ? null : new MicroAligner3(index, minIdentity, true));
if(dumpIn==null){
flowcell=new FlowCell(k);
if(loadKmers){loadKmers();}
fillTiles();
keySets=null;
bloomFilter=null;
bloomCorrector=null;
}else{
flowcell=new FlowCell(dumpIn);
if(flowcell.avgReads<targetAverageReads){
flowcell=flowcell.widenToTargetReads(targetAverageReads);
}
}
Shared.setThreads(oldThreads);
}
/** Create read streams and process all data */
void loadKmers(){
Timer t2=new Timer();
outstream.print("Loading kmers: \t");
if(bloomMode) {
BloomFilter.printMem=false;
KmerCountAbstract.KMERS_PER_READ=kmersPerRead;
}
if(bloomMode && multithreadedLoad) {
KmerCountAbstract.CANONICAL=true;
// ReadCounter rc=new ReadCounter(k, true, false, true, false);
bloomFilter=new BloomFilter(in1, in2, extra, k, k, cbits, hashes, 1, true, false, true, 0.7f);
}else {
//Create a read input stream
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, ffin2);
cris.start(); //Start the stream
if(verbose){outstream.println("Started cris");}
}
boolean paired=cris.paired();
// if(!ffin1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
//Process the read stream
loadKmersInner(cris);
if(verbose){outstream.println("Finished; closing streams.");}
//Close the read streams
errorState|=ReadWrite.closeStreams(cris);
}
t2.stop();
outstream.println(t2);
if(bloomFilter!=null) {
bloomCorrector=new BloomFilterCorrector(bloomFilter, k, k);
double used=bloomFilter.filter.usedFraction();
long unique=(long)bloomFilter.filter.estimateUniqueKmersFromUsedFraction(hashes, used);
System.err.println(String.format("Bloom Occupancy:\t%.2f%%", 100*used));
System.err.println("Unique Kmers: \t"+unique);
// System.err.println("Cells: \t"+bloomFilter.filter.cells);
// System.err.println("Cells Used: \t"+bloomFilter.filter.cellsUsed());
assert(keySets==null);
BloomFilter.printMem=true;
}
KmerCountAbstract.KMERS_PER_READ=0;
}
BarcodeStats loadBarcodes(String expectedBarcodesFile) {
if(delimiter<0) {
delimiter=(byte)ffin1.barcodeDelimiter();
barcodesPerRead=ffin1.barcodesPerRead();
}
BarcodeStats bs=new BarcodeStats(delimiter, barcodesPerRead, extin);
if(bs.length1<1 && bs.length2<1) {
bs.length1=ffin1.barcodeLength(1);
bs.length2=ffin1.barcodeLength(2);
}
if(expectedBarcodesFile!=null) {
bs.loadBarcodeList(expectedBarcodesFile, barcodesPerRead>1 ? delimiter : 0, false, false);
}
return bs;
}
/** Create read streams and process all data */
void fillTiles(){
//Create a read input stream
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, ffin2);
cris.start(); //Start the stream
if(verbose){outstream.println("Started cris");}
}
// boolean paired=cris.paired();
// if(!ffin1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
Timer t2=new Timer();
//Process the read stream
outstream.print("Filling tiles: \t");
if(multithreadedFill) {
spawnThreads(cris);
}else {
fillTilesInner(cris);
}
t2.stop();
outstream.println(t2);
ArrayList<MicroTile> mtList=flowcell.calcStats();
if(flowcell.avgReads<targetAverageReads){
flowcell=flowcell.widenToTargetReads(targetAverageReads);
mtList=flowcell.toList();
}
if(dump!=null){
flowcell.dump(dump, overwrite);
}
if(verbose){outstream.println("Finished; closing streams.");}
//Close the read streams
errorState|=ReadWrite.closeStreams(cris);
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
/** Iterate through the reads */
private void loadKmersInner(final ConcurrentReadInputStream cris){
if(bloomMode) {
bloomFilter=new BloomFilter(k, k, cbits, hashes, 1, true, false, true, 0.7f);
}else {
keySets=new AbstractKmerTable[WAYS];
//Initialize tables
ScheduleMaker scheduleMaker=new ScheduleMaker(WAYS, 12, false, 0.8);
int[] schedule=scheduleMaker.makeSchedule();
for(int j=0; j<WAYS; j++){
keySets[j]=new HashArray1D(schedule, -1L);
}
}
//Do anything necessary prior to processing
loadKmersST(cris);
if(bloomMode) {bloomFilter.filter.shutdown();}
//Do anything necessary after processing
}
private void loadKmersST(final ConcurrentReadInputStream cris) {
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Grab the actual read list from the ListNum
ArrayList<Read> reads=(ln!=null ? ln.list : null);
//Check to ensure pairing is as expected
if(reads!=null && !reads.isEmpty()){
Read r=reads.get(0);
assert((ffin1==null || ffin1.samOrBam()) || (r.mate!=null)==cris.paired());
}
LongList kmers=new LongList(300);
//As long as there is a nonempty read list...
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
if(verbose){outstream.println("Fetched "+reads.size()+" reads.");}
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
final Read r2=r1.mate;
loadKmers(r1, kmers);
loadKmers(r2, kmers);
}
//Notify the input stream that the list was used
cris.returnList(ln);
if(verbose){outstream.println("Returned a list.");}
//Fetch a new list
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
private void loadKmers(Read r, LongList kmers) {
if(r==null || r.length()<k) {return;}
// if(!randy.nextBoolean()) {return;}//Speed optimization, I guess
if(kmersPerRead<1) {
kmers.clear();
BloomFilter.toKmers(r, kmers, k, 0, 0, true);
if(keySets!=null) {
for(int i=0; i<kmers.size; i++) {
long kmer=kmers.array[i];
AbstractKmerTable table=keySets[(int)(kmer%WAYS)];
table.increment(kmer, 1);
}
}else {
for(int i=0; i<kmers.size; i++) {
long kmer=kmers.array[i];
bloomFilter.filter.increment(kmer);
}
}
}else {
// final long kmer=toKmer(r.bases, randy.nextInt(r.length()-k2), k);
final long kmer=toKmer(r.bases, (int)(r.numericID%(r.length()-k2)), k);
if(kmer>=0){
if(keySets!=null) {
AbstractKmerTable table=keySets[(int)(kmer%WAYS)];
table.increment(kmer, 1);
}else {
bloomFilter.filter.increment(kmer);
}
}
}
}
/** Iterate through the reads */
private void fillTilesInner(final ConcurrentReadInputStream cris){
LongList kmers=new LongList();
IntList counts=new IntList();
{
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Grab the actual read list from the ListNum
ArrayList<Read> reads=(ln!=null ? ln.list : null);
//Check to ensure pairing is as expected
if(reads!=null && !reads.isEmpty()){
Read r=reads.get(0);
assert((ffin1==null || ffin1.samOrBam()) || (r.mate!=null)==cris.paired());
}
//As long as there is a nonempty read list...
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
if(verbose){outstream.println("Fetched "+reads.size()+" reads.");}
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
final Read r2=r1.mate;
//Track the initial length for statistics
final int initialLength1=r1.length();
final int initialLength2=r1.mateLength();
//Increment counters
readsProcessed+=r1.pairCount();
basesProcessed+=initialLength1+initialLength2;
final MicroTile mt=flowcell.getMicroTile(r1.id);
if(loadKmers){
processTileKmers(r1, mt, kmers, counts);
processTileKmers(r2, mt, kmers, counts);
}
if(mapper!=null) {
mapper.map(r1);
mapper.map(r2);
}
if(barcodeStats!=null) {
String code=r1.barcode(true);
int hdist=barcodeStats.calcHdist(code);
mt.barcodes+=barcodesPerRead;
mt.barcodeHDistSum+=hdist;
}
mt.add(r1);
mt.add(r2);
}
//Notify the input stream that the list was used
cris.returnList(ln);
if(verbose){outstream.println("Returned a list.");}
//Fetch a new list
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
}
void processTileKmers(Read r, MicroTile mt, LongList kmers, IntList counts) {
if(r==null || r.length()<k) {return;}
if(kmersPerRead<1 && bloomMode) {
processAllKmers(r, mt, kmers, counts);
}else {
processOneKmer(r, mt);
}
}
void processOneKmer(Read r, MicroTile mt) {
// final long kmer=toKmer(r.bases, randy.nextInt(r.length()-k2), k);
final long kmer=toKmer(r.bases, (int)((r.numericID+1)%(r.length()-k2)), k);
if(kmer<0) {
mt.misses++;
return;
}
final long key=(kmersPerRead<1 ? Tools.max(kmer, AminoAcid.reverseComplementBinaryFast(kmer, k)) : kmer);
final int value=(keySets==null ? bloomFilter.getCount(key) : keySets[(int)(key%WAYS)].getValue(key));
mt.depthSum+=value;
final int cutoff=(kmersPerRead<1 ? 2 : 1);
if(value>=cutoff) {mt.hits++;}
else {mt.misses++;}
}
void processAllKmers(Read r, MicroTile mt, LongList kmers, IntList counts) {
final byte[] bases=r.bases;
final byte[] quals=r.quality;
int valid=bloomCorrector.fillKmers(bases, kmers);
if(valid<2){return;}
// if(!r.containsUndefined() && !bloomCorrector.hasErrorsFast(kmers)){return;}
bloomCorrector.fillCounts(bases, kmers, counts);
final int depth=(counts.size<1 ? 0 : counts.get((int)((r.numericID+1)%counts.size)));
mt.depthSum+=depth;
if(depth>1) {mt.hits++;}
else {mt.misses++;}
final int possibleErrors=bloomCorrector.countErrors(counts, quals);
mt.kmerBaseErrorCount+=possibleErrors;
mt.kmerReadErrorCount+=(possibleErrors>0 ? 1 : 0);
}
/*--------------------------------------------------------------*/
/*---------------- Threading ----------------*/
/*--------------------------------------------------------------*/
/** Spawn process threads */
private void spawnThreads(final ConcurrentReadInputStream cris){
//Do anything necessary prior to processing
//Determine how many threads may be used
final int threads=Shared.threads();
//Fill a list with ProcessThreads
ArrayList<WorkerThread> alpt=new ArrayList<WorkerThread>(threads);
for(int i=0; i<threads; i++){
alpt.add(new WorkerThread(cris, i));
}
//Start the threads and wait for them to finish
boolean success=ThreadWaiter.startAndWait(alpt, this);
errorState&=!success;
//Do anything necessary after processing
}
@Override
public final void accumulate(WorkerThread pt){
// System.err.println("Accumulating "+pt.tid+": "+pt.readsProcessedT);
synchronized(pt) {
flowcell.add(pt.fc);
readsProcessed+=pt.readsProcessedT;
basesProcessed+=pt.basesProcessedT;
errorState|=(!pt.success);
}
// System.err.println("readsProcessed: "+readsProcessed+", "+flowcell.readsProcessed);
// pt.fc.dump(pt.tid+".txt", overwrite);
}
@Override
public final boolean success() {return !errorState;}
class WorkerThread extends Thread {
WorkerThread(ConcurrentReadInputStream cris_, int tid_){
cris=cris_;
tid=tid_;
fc=new FlowCell(k);
}
//Called by start()
@Override
public void run(){
//Do anything necessary prior to processing
//Process the reads
processInner();
//Do anything necessary after processing
//Indicate successful exit status
success=true;
}
/** Iterate through the reads */
void processInner(){
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//As long as there is a nonempty read list...
while(ln!=null && ln.size()>0){
// if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
processList(ln);
//Notify the input stream that the list was used
cris.returnList(ln);
// if(verbose){outstream.println("Returned a list.");} //Disabled due to non-static access
//Fetch a new list
ln=cris.nextList();
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
void processList(ListNum<Read> ln){
//Grab the actual read list from the ListNum
final ArrayList<Read> reads=ln.list;
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
final Read r2=r1.mate;
//Validate reads in worker threads
if(!r1.validated()){r1.validate(true);}
if(r2!=null && !r2.validated()){r2.validate(true);}
{
//Reads are processed in this block.
processReadPair(r1, r2);
}
}
//Output reads to the output stream
// if(ros!=null){ros.add(reads, ln.id);}
}
/**
* Process a read or a read pair.
* @param r1 Read 1
* @param r2 Read 2 (may be null)
* @return True if the reads should be kept, false if they should be discarded.
*/
void processReadPair(final Read r1, final Read r2){
//Increment counters
readsProcessedT+=r1.pairCount();
basesProcessedT+=r1.pairLength();
final MicroTile mt=fc.getMicroTile(r1.id);
if(loadKmers){
processTileKmers(r1, mt, kmers, counts);
processTileKmers(r2, mt, kmers, counts);
}
if(mapper!=null) {
mapper.map(r1);
mapper.map(r2);
}
if(barcodeStats!=null) {
String code=r1.barcode(true);
int hdist=barcodeStats.calcHdist(code);
mt.barcodes+=barcodesPerRead;
mt.barcodeHDistSum+=hdist;
}
mt.add(r1);
mt.add(r2);
}
/** Number of reads processed by this thread */
protected long readsProcessedT=0;
/** Number of bases processed by this thread */
protected long basesProcessedT=0;
/** True only if this thread has completed successfully */
boolean success=false;
/** Shared input stream */
private final ConcurrentReadInputStream cris;
/** Thread ID */
final int tid;
final FlowCell fc;
final LongList kmers=new LongList();
final IntList counts=new IntList();
}
/*--------------------------------------------------------------*/
/*---------------- Helper Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* Generate a kmer from specified start location
* @param bases
* @param start
* @param klen kmer length
* @return kmer
*/
private static final long toKmer(final byte[] bases, final int start, final int klen){
final int stop=start+klen;
assert(stop<=bases.length) : klen+", "+bases.length;
long kmer=0;
for(int i=start; i<stop; i++){
final byte b=bases[i];
final long x=Dedupe.baseToNumber[b];
kmer=((kmer<<2)|x);
}
return kmer;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file path */
private String in1=null;
/** Secondary input file path */
private String in2=null;
private ArrayList<String> extra=new ArrayList<String>();
/** Override input file extension */
private String extin=null;
private boolean pound=true;
private String dump=null;
private String dumpIn=null;
/*--------------------------------------------------------------*/
/** Number of reads processed */
public long readsProcessed=0;
/** Number of bases processed */
public long basesProcessed=0;
/** Quit after processing this many input reads; -1 means no limit */
private long maxReads=-1;
/** Hold kmers. A kmer X such that X%WAYS=Y will be stored in keySets[Y] */
private AbstractKmerTable[] keySets;
private BloomFilter bloomFilter;
private BloomFilterCorrector bloomCorrector;
private boolean bloomMode=false;
private boolean multithreadedLoad=false;
private boolean multithreadedFill=false;
private boolean loadKmers=true;
private int kmersPerRead=1;
private int cbits=4;
private int hashes=3;
private int targetAverageReads=800;
private static final int WAYS=31;
private static final int k=31, k2=30;
// private final Random randy=Shared.threadLocalRandom();
private FlowCell flowcell;
private String refPath="phix";
// private byte[] ref;
// private LongHashMap refIndex;
private MicroAligner3 mapper;
float minIdentity=0.65f;
int alignK=19; //Phix is unique down to k=13
private byte delimiter='+';
private int barcodesPerRead=2;
private String expectedBarcodes;
private BarcodeStats barcodeStats;
// private long minCountToUse=0;
//
// private boolean warned=false;
/*--------------------------------------------------------------*/
/*---------------- Final Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file */
private final FileFormat ffin1;
/** Secondary input file */
private final FileFormat ffin2;
@Override
public final ReadWriteLock rwlock() {return rwlock;}
private final ReadWriteLock rwlock=new ReentrantReadWriteLock();
/*--------------------------------------------------------------*/
/*---------------- Common Fields ----------------*/
/*--------------------------------------------------------------*/
/** Print status messages to this output stream */
private PrintStream outstream=System.err;
/** Print verbose messages */
public static boolean verbose=false;
/** True if an error was encountered */
public boolean errorState=false;
/** Overwrite existing output files */
private boolean overwrite=true;
/** Append to existing output files */
private boolean append=false;
}
|