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 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258 259 260 261 262 263 264 265 266 267 268 269 270 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 286 287 288 289 290 291 292 293 294 295 296 297 298 299 300 301 302 303 304 305 306 307 308 309 310 311 312 313 314 315 316 317 318 319 320 321 322 323 324 325 326 327 328 329 330 331 332 333 334 335 336 337 338 339 340 341 342 343 344 345 346 347 348 349 350 351 352 353 354 355 356 357 358 359 360 361 362 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396 397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412 413 414 415 416 417 418 419 420 421 422 423 424 425 426 427 428 429 430 431 432 433 434 435 436 437 438 439 440 441 442 443 444 445 446 447 448 449 450 451 452 453 454 455 456 457 458 459 460 461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592 593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626 627 628 629 630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664 665 666 667 668 669 670 671 672 673 674 675 676 677 678 679 680 681 682 683 684 685 686 687 688 689 690 691 692 693 694 695 696 697 698 699 700 701 702 703 704 705 706 707 708 709 710 711 712 713 714 715 716 717 718 719 720 721 722 723 724 725 726 727 728 729 730 731 732 733 734 735 736 737 738 739 740 741 742 743 744 745 746 747 748 749 750 751 752 753 754 755 756 757 758 759 760 761 762 763 764 765 766 767 768 769 770 771 772 773 774 775 776 777 778 779 780 781 782 783 784 785 786 787 788 789 790 791 792 793 794 795 796 797 798 799 800 801 802 803 804 805 806 807 808 809 810 811 812 813 814 815 816 817 818 819 820 821 822 823 824 825 826 827 828 829 830 831 832 833 834 835 836 837 838 839 840 841 842 843 844 845 846 847 848 849 850 851 852 853 854 855 856 857 858 859 860 861 862 863 864 865 866 867 868 869 870 871 872 873 874 875 876 877 878 879 880 881 882 883 884 885 886 887 888 889 890 891 892 893 894 895 896 897 898 899 900 901 902 903 904 905 906 907 908 909 910 911 912 913 914 915 916 917 918 919 920 921 922 923 924 925 926 927 928 929 930 931 932 933 934 935 936 937 938 939 940 941 942 943 944 945 946 947 948 949 950 951 952 953 954 955 956 957 958 959 960 961 962 963 964 965 966 967 968 969 970 971 972 973 974 975 976 977 978 979 980 981 982 983 984 985 986 987 988 989 990 991 992 993 994 995 996 997 998 999 1000 1001 1002 1003 1004 1005 1006 1007 1008 1009 1010 1011 1012 1013 1014 1015 1016 1017 1018 1019 1020 1021 1022 1023 1024 1025 1026 1027 1028 1029 1030 1031 1032 1033 1034 1035 1036 1037 1038 1039 1040 1041 1042 1043 1044 1045 1046 1047 1048 1049 1050 1051 1052 1053 1054 1055 1056 1057 1058 1059 1060 1061 1062 1063 1064 1065 1066 1067 1068 1069 1070 1071 1072 1073 1074 1075 1076 1077 1078 1079 1080 1081 1082 1083 1084 1085 1086 1087 1088 1089 1090 1091 1092 1093 1094 1095 1096 1097 1098 1099 1100 1101 1102 1103 1104 1105 1106 1107 1108 1109 1110 1111 1112 1113 1114 1115 1116 1117 1118 1119 1120 1121 1122 1123 1124 1125 1126 1127 1128 1129 1130 1131 1132 1133 1134 1135 1136 1137 1138 1139 1140 1141 1142 1143 1144 1145 1146 1147 1148 1149 1150 1151 1152 1153 1154 1155 1156 1157 1158 1159 1160 1161 1162 1163 1164 1165 1166 1167 1168 1169 1170 1171 1172 1173 1174 1175 1176 1177 1178 1179 1180 1181 1182 1183 1184 1185 1186 1187 1188 1189 1190 1191 1192 1193 1194 1195 1196 1197 1198 1199 1200 1201 1202 1203 1204 1205 1206 1207 1208 1209 1210 1211 1212 1213 1214 1215 1216 1217 1218 1219 1220 1221 1222 1223 1224 1225 1226 1227 1228 1229 1230 1231 1232 1233 1234 1235 1236 1237 1238 1239 1240 1241 1242 1243 1244 1245 1246 1247 1248 1249 1250 1251 1252 1253
|
package jgi;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Arrays;
import java.util.BitSet;
import java.util.HashMap;
import java.util.LinkedHashMap;
import java.util.concurrent.ArrayBlockingQueue;
import java.util.concurrent.ConcurrentHashMap;
import dna.AminoAcid;
import fileIO.ByteFile;
import fileIO.ByteFile1;
import fileIO.ByteFile2;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.KillSwitch;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentGenericReadInputStream;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import stream.SamLine;
import structures.CoverageArray;
import structures.CoverageArray2;
import structures.CoverageArray3;
import structures.IntList;
import structures.ListNum;
import tracker.EntropyTracker;
import tracker.ReadStats;
/**
* Masks a fasta file by inserting 'N' in place of low-complexity short repeats,
* and anything covered by mapped reads in a sam file.
*
* @author Brian Bushnell
* @date Feb 18, 2014
*
*/
public class BBMask{
public static void main(String[] args){
Timer t=new Timer();
BBMask x=new BBMask(args);
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
public BBMask(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(16);
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=false;
boolean setEntropyMode=false;
for(int i=0; i<args.length; i++){
String arg=args[i];
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(Parser.parseCommonStatic(arg, a, b)){
//do nothing
}else if(Parser.parseZip(arg, a, b)){
//do nothing
}else if(Parser.parseQuality(arg, a, b)){
//do nothing
}else if(Parser.parseFasta(arg, a, b)){
//do nothing
}else if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
ByteFile1.verbose=verbose;
ByteFile2.verbose=verbose;
stream.FastaReadInputStream.verbose=verbose;
ConcurrentGenericReadInputStream.verbose=verbose;
stream.FastqReadInputStream.verbose=verbose;
ReadWrite.verbose=verbose;
}else if(a.equals("reads") || a.equals("maxreads")){
maxReads=Parse.parseKMG(b);
}else if(a.equals("t") || a.equals("threads")){
Shared.setThreads(b);
}else if(a.equals("sampad") || a.equals("sampadding") || a.equals("sp")){
samPad=Integer.parseInt(b);
}else if(a.equals("entropymode")){
entropyMode=Parse.parseBoolean(b);
setEntropyMode=true;
}else if(a.equals("maskrepeats") || a.equals("mr")){
processRepeats=Parse.parseBoolean(b);
}else if(a.equals("masklowentropy") || a.equals("masklowcomplexity") || a.equals("mlc") || a.equals("mle") || a.equals("me")){
processEntropy=Parse.parseBoolean(b);
}else if(a.equals("in") || a.equals("input") || a.equals("in1") || a.equals("input1") || a.equals("ref")){
inRef=b;
}else if(a.equals("insam") || a.equals("samin") || a.equals("sam")){
if(b!=null){
for(String s : b.split(",")){
inSam.add(s);
}
}
}else if(a.equals("out") || a.equals("output") || a.equals("out1") || a.equals("output1") || a.equals("output1")){
outRef=b;
}else if(a.equals("qfin") || a.equals("qfin1")){
qfinRef=b;
}else if(a.equals("qfout") || a.equals("qfout1")){
qfoutRef=b;
}else if(a.equals("extin")){
extinRef=b;
}else if(a.equals("extout")){
extoutRef=b;
}else if(a.equals("split")){
splitMode=Parse.parseBoolean(b);
}else if(a.equals("mink") || a.equals("kmin")){
mink=mink2=Integer.parseInt(b);
}else if(a.equals("maxk") || a.equals("kmax")){
maxk=maxk2=Integer.parseInt(b);
}else if(a.equals("k")){
mink=maxk=mink2=maxk2=Integer.parseInt(b);
}else if(a.equals("minkr") || a.equals("krmin")){
mink=Integer.parseInt(b);
}else if(a.equals("maxkr") || a.equals("krmax")){
maxk=Integer.parseInt(b);
}else if(a.equals("kr")){
mink=maxk=Integer.parseInt(b);
}else if(a.equals("mink2") || a.equals("kmin2") || a.equals("minke") || a.equals("kemin")){
mink2=Integer.parseInt(b);
}else if(a.equals("maxk2") || a.equals("kmax2") || a.equals("maxke") || a.equals("kemax")){
maxk2=Integer.parseInt(b);
}else if(a.equals("k2") || a.equals("ke")){
mink2=maxk2=Integer.parseInt(b);
}else if(a.equals("mincov")){
mincov=Integer.parseInt(b);
}else if(a.equals("maxcov")){
maxcov=Integer.parseInt(b);
}else if(a.equals("delcov") || a.equals("delcoverage")){
includeDeletionCoverage=Parse.parseBoolean(b);
}else if(a.equals("window") || a.equals("w")){
window=Integer.parseInt(b);
}else if(a.equals("ratio")){
ratio=Float.parseFloat(b);
if(!setEntropyMode){entropyMode=false;}
}else if(a.equals("entropy") || a.equals("e")){
entropyCutoff=Float.parseFloat(b);
if(!setEntropyMode){entropyMode=true;}
}else if(a.equals("lowercase") || a.equals("lc")){
MaskByLowercase=Parse.parseBoolean(b);
}else if(a.equals("minlen")){
minlen=Integer.parseInt(b);
}else if(a.equals("mincount")){
mincount=Integer.parseInt(b);
}else if(a.equals("append") || a.equals("app")){
append=ReadStats.append=Parse.parseBoolean(b);
}else if(a.equals("overwrite") || a.equals("ow")){
overwrite=Parse.parseBoolean(b);
}else if(a.startsWith("minscaf") || a.startsWith("mincontig")){
stream.FastaReadInputStream.MIN_READ_LEN=Integer.parseInt(b);
}else if(!arg.contains("=") && FileFormat.hasSamOrBamExtension(arg)){
inSam.add(arg);
}
else if(inRef==null && i==0 && Tools.looksLikeInputStream(arg)){
inRef=arg;
}
// else if(outRef==null && i==1 && !arg.contains("=")){
// outRef=arg;
// }
else{
System.err.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
// throw new RuntimeException("Unknown parameter "+args[i]);
}
}
bits32=(mincov>=Character.MAX_VALUE || maxcov>=Character.MAX_VALUE);
{//Process parser fields
Parser.processQuality();
}
assert(FastaReadInputStream.settingsOK());
if(inRef==null){throw new RuntimeException("Error - at least one input file is required.");}
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
if(outRef!=null && outRef.equalsIgnoreCase("null")){outRef=null;}
if(!Tools.testOutputFiles(overwrite, append, false, outRef)){
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output file "+outRef+"\n");
}
ffoutRef=FileFormat.testOutput(outRef, FileFormat.FASTA, extoutRef, true, overwrite, append, false);
ffinRef=FileFormat.testInput(inRef, FileFormat.FASTA, extinRef, true, true);
if(inSam!=null && inSam.size()>0){
ffinSam=new FileFormat[inSam.size()];
for(int i=0; i<inSam.size(); i++){
ffinSam[i]=FileFormat.testInput(inSam.get(i), FileFormat.SAM, null, true, false);
}
}else{
ffinSam=null;
}
SamLine.CONVERT_CIGAR_TO_MATCH=false;
EntropyTracker.verify=verify;
}
/*--------------------------------------------------------------*/
public void process(Timer t0){
Timer t=new Timer();
{
t.start();
outstream.println("Loading input");
boolean oldTI=FASTQ.TEST_INTERLEAVED;
boolean oldFI=FASTQ.FORCE_INTERLEAVED;
FASTQ.TEST_INTERLEAVED=false;
FASTQ.FORCE_INTERLEAVED=false;
map=hashRef();
FASTQ.TEST_INTERLEAVED=oldTI;
FASTQ.FORCE_INTERLEAVED=oldFI;
t.stop();
outstream.println("Loading Time: \t"+t);
}
long repeats=0, lowcomplexity=0;
long mapping=0;
if(processRepeats && maxk>0){
t.start();
outstream.println("\nMasking repeats (to disable, set 'mr=f')");
// repeats=maskRepeats_ST();
repeats=maskRepeats();
t.stop();
double rpnano=refReads/(double)(t.elapsed);
double bpnano=refBases/(double)(t.elapsed);
String rpstring=""+refReads;
String bpstring=""+refBases;
String bmstring=""+repeats;
while(rpstring.length()<12){rpstring=" "+rpstring;}
while(bpstring.length()<12){bpstring=" "+bpstring;}
while(bmstring.length()<12){bmstring=" "+bmstring;}
outstream.println("Repeat Masking Time: \t"+t);
//outstream.println("Ref Scaffolds: "+rpstring+" \t"+Tools.format("%.2fk scafs/sec", rpnano*1000000));
outstream.println("Ref Bases: "+bpstring+" \t"+Tools.format("%.2fm bases/sec", bpnano*1000));
outstream.println("Repeat Bases Masked: "+bmstring);
}
if(processEntropy && maxk2>0){
t.start();
if(entropyMode){
outstream.println("\nMasking low-entropy (to disable, set 'mle=f')");
lowcomplexity=maskLowEntropy();
}else{
outstream.println("\nMasking low-complexity (to disable, set 'mlc=f')");
lowcomplexity=maskLowComplexity(null);
}
t.stop();
double rpnano=refReads/(double)(t.elapsed);
double bpnano=refBases/(double)(t.elapsed);
String rpstring=""+refReads;
String bpstring=""+refBases;
String bmstring=""+lowcomplexity;
while(rpstring.length()<12){rpstring=" "+rpstring;}
while(bpstring.length()<12){bpstring=" "+bpstring;}
while(bmstring.length()<12){bmstring=" "+bmstring;}
outstream.println("Low Complexity Masking Time: \t"+t);
//outstream.println("Ref Scaffolds: "+rpstring+" \t"+Tools.format("%.2fk scafs/sec", rpnano*1000000));
outstream.println("Ref Bases: "+bpstring+" \t"+Tools.format("%.2fm bases/sec", bpnano*1000));
outstream.println("Low Complexity Bases: "+bmstring);
}
if(ffinSam!=null){
t.start();
outstream.println("\nMasking from sam");
mapping=maskSam();
t.stop();
double rpnano=samReads/(double)(t.elapsed);
double bpnano=samBases/(double)(t.elapsed);
String rpstring=""+samReads;
String bpstring=""+samBases;
String bmstring=""+mapping;
while(rpstring.length()<12){rpstring=" "+rpstring;}
while(bpstring.length()<12){bpstring=" "+bpstring;}
while(bmstring.length()<12){bmstring=" "+bmstring;}
outstream.println("Sam Masking Time: \t"+t);
outstream.println("Sam Reads Processed: "+rpstring+" \t"+Tools.format("%.2fk reads/sec", rpnano*1000000));
outstream.println("Sam Bases Processed: "+bpstring+" \t"+Tools.format("%.2fm bases/sec", bpnano*1000));
outstream.println("Sam Bases Masked: "+bmstring);
}
long total=repeats+mapping+lowcomplexity, masked=0;
if(total>0 || true){
t.start();
if(splitMode){
masked=splitFromBitsets();
}else{
masked=maskFromBitsets(MaskByLowercase);
}
t.stop();
outstream.println("Conversion Time: \t"+t);
}
assert(total==masked) : repeats+", "+mapping+", "+lowcomplexity+", "+total+", "+masked;
if(outRef!=null){
t.start();
outstream.println("\nWriting output");
writeOutput();
t.stop();
outstream.println("Writing Time: \t"+t);
}
{
t0.stop();
String tstring=""+total;
while(tstring.length()<12){tstring=" "+tstring;}
outstream.println("\nTotal Bases Masked: "+tstring+"/"+refBases+Tools.format("\t%.3f%%", total*100.0/refBases));
outstream.println("Total Time: \t"+t0);
}
if(errorState){
throw new RuntimeException("\nBBMask terminated in an error state; the output may be corrupt.");
}
}
/*--------------------------------------------------------------*/
private static int setHighCoverage(BitSet bs, CoverageArray ca, int maxAllowedCoverage, int maxLen){
int numSet=0;
for(int i=0; i<maxLen; i++){
if(ca.get(i)>maxAllowedCoverage){
bs.set(i);
numSet++;
}
}
return numSet;
}
private static int setLowCoverage(BitSet bs, CoverageArray ca, int minAllowedCoverage, int maxLen){
int numSet=0;
for(int i=0; i<maxLen; i++){
if(ca.get(i)<minAllowedCoverage){
bs.set(i);
numSet++;
}
}
return numSet;
}
private long maskFromBitsets(final boolean lowercase){
System.err.println("\nConverting masked bases to "+(lowercase ? "lower case" : "N")); //123
long sum=0;
for(Read r : map.values()){
sum+=maskRead(r, (BitSet)r.obj, lowercase);
}
System.err.println("Done Masking");
return sum;
}
public static int maskRead(final Read r, final BitSet bs, final boolean lowercase){
final byte[] bases=r.bases;
int sum=0;
if(!lowercase){
for(int i=0; i<bases.length; i++){
if(bs.get(i)){
if(bases[i]!='N'){sum++;}
bases[i]='N';
}else if(CONVERT_NON_ACGTN && !AminoAcid.isACGTN(bases[i])){
bases[i]='N';
}
}
}else{
for(int i=0; i<bases.length; i++){
if(bs.get(i)){
if(!Tools.isLowerCase(bases[i]) && bases[i]!='N'){sum++;}
bases[i]=(byte)Tools.toLowerCase(bases[i]);
}else if(CONVERT_NON_ACGTN && !AminoAcid.isACGTN(bases[i])){
bases[i]='N';
}
}
}
return sum;
}
private long splitFromBitsets(){
System.err.println("\nSplitting reads by removing masked bases"); //123
long sum=0;
LinkedHashMap<String, Read> map2=new LinkedHashMap<String, Read>();
for(String key : map.keySet()){
Read r=map.get(key);
BitSet bs=((BitSet)r.obj);
int rnum=0;
if(bs.isEmpty()){
map2.put(key, r);
}else{
byte[] bases=r.bases;
byte[] quals=r.quality;
int lastGood, lastBad;
if(bs.get(0)){
lastGood=-1;
lastBad=0;
}else{
lastGood=0;
lastBad=-1;
}
int i=1;
for(; i<bases.length; i++){
if(bs.get(i)){
if(lastGood==i-1){
int len=lastGood-lastBad;
if(len>0){
byte[] bases2=KillSwitch.copyOfRange(bases, lastBad+1, i);
byte[] quals2=(quals==null ? null : KillSwitch.copyOfRange(quals, lastBad+1, i));
Read r2=new Read(bases2, quals2, r.id+"_"+rnum, r.numericID, r.flags);
Read old=map2.put(r2.id, r2);
assert(old==null) : "Duplicate id "+r2.id; //TODO: This can easily be resolved by making a new ID string.
}
}
lastBad=i;
}else{
lastGood=i;
}
}
if(lastGood==i-1){
int len=lastGood-lastBad;
if(len>0){
byte[] bases2=KillSwitch.copyOfRange(bases, lastBad+1, i);
byte[] quals2=(quals==null ? null : KillSwitch.copyOfRange(quals, lastBad+1, i));
Read r2=new Read(bases2, quals2, r.id+"_"+rnum, r.numericID, r.flags);
Read old=map2.put(r2.id, r2);
assert(old==null) : "Duplicate id "+r2.id; //TODO: This can easily be resolved by making a new ID string.
}
}
}
}
map.clear();
map.putAll(map2);
map2.clear();
System.err.println("Done Splitting");
return sum;
}
private void writeOutput(){
ConcurrentReadOutputStream ros=null;
if(ffoutRef!=null){
final int buff=16;
ros=ConcurrentReadOutputStream.getStream(ffoutRef, null, qfoutRef, null, buff, null, false);
ros.start();
}
long i=0;
for(String name : map.keySet()){
Read r=map.get(name);
ArrayList<Read> list=new ArrayList<Read>(1);
list.add(r);
if(ros!=null){ros.add(list, i);}
i++;
}
errorState|=ReadWrite.closeStream(ros);
}
/*--------------------------------------------------------------*/
private long maskSam(){
long before=0, after=0;
for(Read r : map.values()){
before+=((BitSet)r.obj).cardinality();
}
for(FileFormat ff : ffinSam){
//maskSam_ST(ff);
maskSam_MT(ff);
}
for(Read r : map.values()){
after+=((BitSet)r.obj).cardinality();
}
return after-before;
}
private void maskSam_MT(FileFormat ff){
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ff, null, null, null);
if(verbose){System.err.println("Started cris");}
cris.start(); //4567
}
MaskSamThread[] threads=new MaskSamThread[Shared.threads()];
// outstream.println("Spawning "+numThreads+" threads.");
for(int i=0; i<threads.length; i++){threads[i]=new MaskSamThread(cris);}
for(int i=0; i<threads.length; i++){threads[i].start();}
for(int i=0; i<threads.length; i++){
while(threads[i].getState()!=Thread.State.TERMINATED){
try {
threads[i].join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
}
if(covmap!=null){//Move coverage array information to BitSets.
for(String rs : map.keySet()){
Read r=map.get(rs);
BitSet bs=(BitSet)r.obj;
CoverageArray ca=covmap.remove(rs);
if(maxcov>-1){
setHighCoverage(bs, ca, maxcov, r.length());
}
if(mincov>-1){
setLowCoverage(bs, ca, mincov, r.length());
}
}
}
errorState|=ReadWrite.closeStreams(cris);
if(errorState){
throw new RuntimeException("BBMask terminated in an error state; the output may be corrupt.");
}
}
private class MaskSamThread extends Thread{
MaskSamThread(ConcurrentReadInputStream cris_){
cris=cris_;
}
@Override
public void run(){
maskSam(cris);
}
final ConcurrentReadInputStream cris;
}
private void maskSam_ST(FileFormat ff){
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ff, null, null, null);
if(verbose){System.err.println("Started cris");}
cris.start(); //4567
}
maskSam(cris);
errorState|=ReadWrite.closeStreams(cris);
if(errorState){
throw new RuntimeException("BBMask terminated in an error state; the output may be corrupt.");
}
}
private void maskSam(ConcurrentReadInputStream cris){
long samReads=0;
long samBases=0;
IntList ranges=new IntList(16);
{
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
for(int idx=0; idx<reads.size(); idx++){
final Read r=reads.get(idx);
assert(r.mate==null);
final int initialLength1=r.length();
{
samReads++;
samBases+=initialLength1;
if(r.mapped()){
SamLine sl=r.samline;
assert(sl!=null) : "No sam line for read "+r;
byte[] rname=sl.rname();
assert(rname!=null) : "No rname for sam line "+sl;
final String rs=new String(rname);
Read ref=map.get(rs);
if(ref==null){
handleNoRef(rs);
}else{
final int reflen=ref.length();
assert(ref!=null) : "Could not find reference scaffold '"+rs+"' for samline \n"+sl+"\n in set \n"+map.keySet()+"\n";
if(covmap==null){
BitSet bs=(BitSet)ref.obj;
mask(bs, sl, reflen);
}else{
CoverageArray ca=covmap.get(rs);
increment(ca, sl, r.match, reflen, ranges, includeDeletionCoverage, samPad);
}
}
}
}
}
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
synchronized(this){
this.samBases+=samBases;
this.samReads+=samReads;
}
}
private void mask(BitSet bs, SamLine sl, int reflen){
final int start=Tools.max(0, sl.start(true, false)-samPad);
final int stop=Tools.min(sl.stop(start, true, false)+1+samPad, reflen);
if(stop>start){
synchronized(bs){//Potential bottleneck
bs.set(start, stop);
}
}
}
public static void increment(CoverageArray ca, SamLine sl, byte[] match, int reflen, IntList ranges, boolean includeDels, int samPad){
final int start=Tools.max(0, sl.start(true, false));
final int stop=Tools.min(sl.stop(start, true, false)+1, reflen);
if(stop>start){
ranges.clear();
boolean numeric=false;
if(match==null){
assert(sl.cigar!=null);
match=sl.toShortMatch(true);
numeric=true;
}else{
for(byte b : match){
if(Tools.isDigit(b)){
numeric=true;
break;
}
}
}
if(numeric){match=Read.toLongMatchString(match);}
fillRanges(match, start, stop, ranges, includeDels);
// assert(false) : ranges;
if(ranges.size>0){
if(samPad!=0){//Pad the ranges, but don't let them overlap
ranges.set(0, Tools.mid(0, reflen, ranges.get(0)-samPad));
ranges.set(1, Tools.mid(0, reflen, ranges.get(1)+samPad));
for(int i=2; i<ranges.size; i+=2){
ranges.set(i, Tools.mid(ranges.get(i-1), reflen, ranges.get(i)-samPad));
ranges.set(i+1, Tools.mid(0, reflen, ranges.get(i+1)+samPad));
}
}
synchronized(ca){//Potential bottleneck
ca.incrementRanges(ranges, 1);
}
}
}
}
public static void fillRanges(byte[] longmatch, int start, int stop, IntList ranges, boolean includeDels){
assert(ranges.size==0);
byte mode='?', lastMode='?';
int rpos=start;
int lastRpos=start;
int rstart=start;
for(int mpos=0; mpos<longmatch.length; mpos++){
byte m=longmatch[mpos];
if(m=='m' || m=='s' || m=='S' || m=='N' || m=='B'){//Little 's' is for a match classified as a sub to improve the affine score.
mode='m';
rpos++;
}else if(m=='I' || m=='X' || m=='Y'){
mode='I';
}else if(m=='D'){
mode='D';
rpos++;
}else if(m=='C'){
mode='C';
rpos++;
}else{
throw new RuntimeException("Invalid match string character '"+(char)m+"' = "+m+" (ascii). " +
"Match string should be in long format here.");
}
if(mode!=lastMode){
if(mpos>0){
if(lastMode=='m'){
ranges.add(rstart);
ranges.add(lastRpos);
}else if(mode=='D' && includeDels){
ranges.add(rstart);
ranges.add(lastRpos);
}
}
rstart=lastRpos;
}
lastMode=mode;
lastRpos=rpos;
}
// assert(false) : rstart+", "+rpos+", "+(char)mode+", "+(char)lastMode;
//Final cycle
if(lastMode=='m'){
ranges.add(rstart);
ranges.add(lastRpos);
}else if(mode=='D' && includeDels){
ranges.add(rstart);
ranges.add(lastRpos);
}
assert(rpos==stop) : start+", "+stop+", "+rpos+", "+rstart+", "+lastRpos+"\n"+new String(longmatch)+"\n"+ranges;
assert((ranges.size&1)==0);
}
private void handleNoRef(String rname){
assert(rname!=null);
String ret=norefSet.putIfAbsent(rname, rname);
if(ret==null){
System.err.println("Warning! Scaffold not found in assembly: "+rname);
}
}
/*--------------------------------------------------------------*/
private LinkedHashMap<String, Read> hashRef(){
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffinRef, null, qfinRef, null);
if(verbose){System.err.println("Started cris");}
cris.start(); //4567
}
final LinkedHashMap<String, Read> hmr=new LinkedHashMap<String, Read>();
if(ffinSam!=null && (mincov>=0 || maxcov>=0)){
covmap=new HashMap<String, CoverageArray>();
}
{
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
for(int idx=0; idx<reads.size(); idx++){
final Read r=reads.get(idx);
final int len=r.length();
final byte[] bases=r.bases;
// refLengths.add(len);
final BitSet bs=new BitSet(len);
r.obj=bs;
if(covmap!=null){
if(bits32){
covmap.put(r.id, new CoverageArray3(covmap.size(), len));
}else{
covmap.put(r.id, new CoverageArray2(covmap.size(), len));
}
}
if(MaskByLowercase){
for(int i=0; i<len; i++){
if(bases[i]=='N' || Tools.isLowerCase(bases[i])){bs.set(i);}
}
}else{
for(int i=0; i<len; i++){
if(bases[i]=='N'){bs.set(i);}
}
}
refReads++;
refBases+=len;
Read old=hmr.put(r.id, r);
assert(old==null) : "Duplicate reference scaffold name "+r.id;
}
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
errorState|=ReadWrite.closeStreams(cris);
if(errorState){
throw new RuntimeException("BBMask terminated in an error state; the output may be corrupt.");
}
return hmr;
}
/*--------------------------------------------------------------*/
private long maskLowComplexity(short[][] matrix){
long sum=0;
if(matrix==null){matrix=new short[16][];}
for(Read r : map.values()){
sum+=maskLowComplexity(r, mink2, maxk2, window, ratio, matrix);
}
return sum;
}
private static int maskLowComplexity(Read r, int mink, int maxk, int window, float ratio, short[][] matrix){
final byte[] bases=r.bases;
final BitSet bs=(BitSet)r.obj;
int before=bs.cardinality();
// System.err.println("\nbefore="+before+"\n"+new String(bases)+"\n"+bs);
for(int k=mink; k<=maxk; k++){
if(matrix[k]==null){matrix[k]=new short[(1<<(2*k))];}
}
for(int k=mink; k<=maxk; k++){
final short[] counts=matrix[k];
final int kmerspace=(1<<(2*k));
final int mincount=(int)Math.ceil(ratio*Tools.min(window, kmerspace));
maskLowComplexity(bases, bs, k, window, mincount, counts);
}
int after=bs.cardinality();
// System.err.println("before="+before+", after="+after+"\n"+new String(bases)+"\n"+bs);
return after-before;
}
private static void maskLowComplexity(final byte[] bases, final BitSet bs, final int k, final int window, final int mincount, final short[] counts){
assert(k>0) : "k must be greater than 0";
if(verify){
for(int c : counts){assert(c==0);}
}
final int mask=(k>15 ? -1 : ~((-1)<<(2*k)));
int current=0, ns=0;
int kmer=0, kmer2=0;
for(int i=0, i2=-window; i2<bases.length; i++, i2++){
// System.err.println("\nStart: i="+i+", current="+current+", ns="+ns+"\n"+Arrays.toString(counts));
if(i<bases.length){
final byte b=bases[i];
final int n=Dedupe.baseToNumber[b];
kmer=((kmer<<2)|n)&mask;
if(!AminoAcid.isFullyDefined(b)){ns++;}
if(counts[kmer]<1){
assert(counts[kmer]==0);
current++;
}
counts[kmer]++;
if(verify){assert(current==Tools.cardinality(counts)) : current+", "+Tools.cardinality(counts)+"\n"+Arrays.toString(counts);}
// System.err.println("Added "+kmer+"; counts["+kmer+"]="+counts[kmer]);
}
if(i2>=0){
final byte b2=bases[i2];
final int n2=Dedupe.baseToNumber[b2];
kmer2=((kmer2<<2)|n2)&mask;
if(!AminoAcid.isFullyDefined(b2)){
ns--;
assert(ns>=0);
}
counts[kmer2]--;
if(counts[kmer2]<1){
assert(counts[kmer2]==0) : Arrays.toString(counts);
current--;
}
if(verify){assert(current==Tools.cardinality(counts)) : current+", "+Tools.cardinality(counts)+"\n"+Arrays.toString(counts);}
// System.err.println("Removed "+kmer2+"; count="+counts[kmer2]);
}
if(verify && i2>-1 && i<bases.length){
assert(shared.Vector.sum(counts)==window);
}
if(current<mincount && ns<1 && i2>=-1 && i<bases.length){
// System.err.println("Masked ("+(i2+1)+", "+(i+1)+")");
bs.set(i2+1, i+1);
}
}
}
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
private long maskLowEntropy(){
ArrayBlockingQueue<Read> queue=new ArrayBlockingQueue<Read>(map.size());
for(Read r : map.values()){queue.add(r);}
int numThreads=Tools.min(Shared.threads(), queue.size());
MaskLowEntropyThread[] threads=new MaskLowEntropyThread[numThreads];
long sum=0;
// outstream.println("Spawning "+numThreads+" threads.");
for(int i=0; i<threads.length; i++){threads[i]=new MaskLowEntropyThread(queue, mink2, maxk2, window, entropyCutoff);}
for(int i=0; i<threads.length; i++){threads[i].start();}
for(int i=0; i<threads.length; i++){
while(threads[i].getState()!=Thread.State.TERMINATED){
try {
threads[i].join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
sum+=threads[i].masked;
}
return sum;
}
private class MaskLowEntropyThread extends Thread{
MaskLowEntropyThread(ArrayBlockingQueue<Read> queue_, int mink_, int maxk_, int window_, float cutoff_){
queue=queue_;
minkT=mink_;
maxkT=maxk_;
windowT=(short)window_;
cutoff=cutoff_;
trackers=new EntropyTracker[maxkT+1];
for(int i=minkT; i<=maxkT; i++){
trackers[i]=new EntropyTracker(i, windowT, false, cutoff, true);
}
}
@Override
public void run(){
for(Read r=queue.poll(); r!=null; r=queue.poll()){
masked+=maskLowEntropy(r, minkT, maxkT, trackers);
}
}
final ArrayBlockingQueue<Read> queue;
final int minkT;
final int maxkT;
final float cutoff;
final int windowT;
final EntropyTracker[] trackers;
long masked=0;
}
private int maskLowEntropy(Read r, int mink, int maxk, EntropyTracker[] trackers){
// outstream.println("maskLowEntropy("+r.numericID+", "+mink+", "+maxk+", "+window+", "+cutoff+", "+matrix.length+", "+countCounts.length+")");
// System.err.println(new String(r.bases));
final byte[] bases=r.bases;
final BitSet bs=(BitSet)r.obj;
int before=bs.cardinality();
for(int k=mink; k<=maxk; k++){
maskLowEntropy(bases, bs, trackers[k]);
}
int after=bs.cardinality();
return after-before;
}
private static void maskLowEntropy(final byte[] bases, final BitSet bs, final EntropyTracker et){
final int window=et.windowBases();
if(bases.length<window){return;}
et.clear();
for(int i=0, min=window-1; i<bases.length; i++){
et.add(bases[i]);
if(i>=min && et.ns()<1 && !et.passes()){bs.set(et.leftPos(), et.rightPos()+1);}
}
}
/*--------------------------------------------------------------*/
private long maskRepeats_ST(){
long sum=0;
for(Read r : map.values()){
sum+=maskRepeats(r, mink, maxk, mincount, minlen);
}
return sum;
}
private long maskRepeats(){
ArrayBlockingQueue<Read> queue=new ArrayBlockingQueue<Read>(map.size());
for(Read r : map.values()){queue.add(r);}
int numThreads=Tools.min(Shared.threads(), queue.size());
MaskRepeatThread[] threads=new MaskRepeatThread[numThreads];
long sum=0;
for(int i=0; i<threads.length; i++){threads[i]=new MaskRepeatThread(queue, mink, maxk, mincount, minlen);}
for(int i=0; i<threads.length; i++){threads[i].start();}
for(int i=0; i<threads.length; i++){
while(threads[i].getState()!=Thread.State.TERMINATED){
try {
threads[i].join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
sum+=threads[i].masked;
}
return sum;
}
private class MaskRepeatThread extends Thread{
MaskRepeatThread(ArrayBlockingQueue<Read> queue_, int mink_, int maxk_, int mincount_, int minlen_){
queue=queue_;
minkT=mink_;
maxkT=maxk_;
mincountT=mincount_;
minlenT=minlen_;
}
@Override
public void run(){
for(Read r=queue.poll(); r!=null; r=queue.poll()){
masked+=maskRepeats(r, minkT, maxkT, mincountT, minlenT);
}
}
final ArrayBlockingQueue<Read> queue;
final int minkT;
final int maxkT;
final int mincountT;
final int minlenT;
long masked=0;
}
private static int maskRepeats(Read r, int mink, int maxk, int mincount, int minlen){
final byte[] bases=r.bases;
final BitSet bs=(BitSet)r.obj;
int before=bs.cardinality();
// System.err.println("\nbefore="+before+"\n"+new String(bases)+"\n"+bs);
for(int k=mink; k<=maxk; k++){
maskRepeats(bases, bs, k, Tools.max(minlen, k*mincount));
}
int after=bs.cardinality();
// System.err.println("before="+before+", after="+after+"\n"+new String(bases)+"\n"+bs);
return after-before;
}
private static void maskRepeats(final byte[] bases, final BitSet bs, final int k, final int minlen){
final int lim=bases.length-k;
final int mask=(k>15 ? -1 : ~((-1)<<(2*k)));
for(int loc=0; loc<lim; loc++){
int len=repeatLength(bases, k, mask, loc);
if(len>=minlen){
int a=loc-k, b=loc-k+len;
bs.set(a, b);
// System.err.println("len="+len+", minlen="+minlen+", set "+(loc-k)+"-"+(loc-k+len));
loc=Tools.max(loc, b-minlen);
// System.err.println("Reset loc to "+loc);
}else{
// System.err.println("len="+len+" < minlen="+minlen);
}
}
}
private static int repeatLength(final byte[] bases, final int k, final int mask, final int loc){
final int lim=bases.length;
final int key=getInitialKey(bases, loc, k);
if(key<0){return 0;}
int kmer=key;
int gap=0, last=-1;
for(int i=loc; i<lim && gap<k; i++){
final byte b=bases[i];
final int n=Dedupe.baseToNumber[b];
kmer=((kmer<<2)|n)&mask;
if(kmer==key){
last=i;
gap=0;
}else{
gap++;
}
// System.err.println("i="+i+", lim="+lim+", gap="+gap+", last="+last+", b="+(char)b+", n="+n+", key="+key+", kmer="+kmer);
}
// System.err.println("k="+k+", mask="+mask+", loc="+loc+", last="+last);
return (last<0 ? 0 : last-loc+k+1);
}
private static int getInitialKey(byte[] bases, int loc, int k){
assert(k<16);
int start=loc-k;
int key=0;
if(start<0){return -1;}
for(int i=start; i<loc; i++){
final byte b=bases[i];
final int n=Dedupe.baseToNumber[b];
key=(key<<2)|n;
}
assert(key>=0);
return key;
}
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
private LinkedHashMap<String, Read> map=null;
private ConcurrentHashMap<String, String> norefSet=new ConcurrentHashMap<String, String>(256, .75f, 16);
private HashMap<String, CoverageArray> covmap=null;
// private IntList refLengths=new IntList();
private long refReads=0;
private long refBases=0;
// private long repeatsMasked=0;
private long samReads=0;
private long samBases=0;
// private long samMasked=0;
public boolean errorState=false;
private String inRef=null;
private ArrayList<String> inSam=new ArrayList<String>();
private String qfinRef=null;
private String outRef=null;
private String qfoutRef=null;
private String extinRef=null;
private String extoutRef=null;
private boolean overwrite=true;
private boolean append=false;
private long maxReads=-1;
private boolean processRepeats=false;
private int mink=5;
private int maxk=5;
private int minlen=40;
private int mincount=4;
private boolean processEntropy=true;
private boolean entropyMode=true;
private boolean splitMode=false;
private int mink2=5;
private int maxk2=5;
private int window=80;
private float ratio=0.35f; //For complexity, if not in entropyMode
private float entropyCutoff=0.70f;
/** Use 32-bit coverage arrays */
private boolean bits32=true;
/** Include deletions when calculating coverage */
private boolean includeDeletionCoverage=true;
/** If nonnegative, mask bases with coverage outside this range. */
private int mincov=-1;
/** If nonnegative, mask bases with coverage outside this range. */
private int maxcov=-1;
private int samPad=0;
private final FileFormat ffinRef;
private final FileFormat[] ffinSam;
private final FileFormat ffoutRef;
private PrintStream outstream=System.err;
/*--------------------------------------------------------------*/
public static boolean verbose=false;
public static boolean CONVERT_NON_ACGTN=true;
private static boolean verify=false;
private static boolean MaskByLowercase=false;
}
|