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package jgi;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Arrays;
import java.util.HashMap;
import java.util.concurrent.ArrayBlockingQueue;
import java.util.concurrent.ConcurrentHashMap;
import java.util.concurrent.atomic.AtomicLongArray;
import barcode.BarcodeMappingStats;
import cardinality.CardinalityTracker;
import dna.AminoAcid;
import dna.Data;
import fileIO.ByteFile;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import fileIO.TextStreamWriter;
import kmer.AbstractKmerTable;
import kmer.ScheduleMaker;
import shared.KillSwitch;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import shared.TrimRead;
import stream.ArrayListSet;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.MultiCros;
import stream.Read;
import stream.SamLine;
import structures.ByteBuilder;
import structures.IntList;
import structures.IntList3;
import structures.ListNum;
import structures.StringCount;
import tax.GiToTaxid;
import tax.TaxNode;
import tax.TaxTree;
import tracker.ReadStats;
/**
* SEAL: Sequence Expression AnaLyzer
* Derived from BBDuk.
* Allows multiple values stored per kmer.
* Intended for RNA-seq, coverage, and other reads-per-sequence quantification.
* Also performs binning.
* @author Brian Bushnell
* @date November 10, 2014
*
*/
public class Seal {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
* @param args Command line arguments
*/
public static void main(String[] args){
//Create a new Seal instance
Seal x=new Seal(args);
///And run it
x.process();
//Close the print stream if it was redirected
Shared.closeStream(outstream);
}
/**
* Constructor.
* @param args Command line arguments
*/
public Seal(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), true);
args=pp.args;
outstream=pp.outstream;
}
/* Set global defaults */
ReadWrite.ZIPLEVEL=2;
ReadWrite.USE_UNPIGZ=true;
ReadWrite.USE_PIGZ=true;
SamLine.SET_FROM_OK=true;
IntList3.defaultMode=IntList3.ASCENDING;
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
/* Initialize local variables with defaults */
boolean rcomp_=true;
boolean forbidNs_=false;
boolean prealloc_=false;
boolean useCountvector_=false;
int tableType_=AbstractKmerTable.ARRAYHF;//Fine as long as numbers are assigned in order per thread; otherwise use ARRAYH.
int k_=31;
int ways_=-1; //Currently disabled
int minKmerHits_=1;
float minKmerFraction_=0;
long skipreads_=0;
Parser parser=new Parser();
parser.trimq=6;
parser.minAvgQuality=0;
parser.minReadLength=10;
parser.maxReadLength=Integer.MAX_VALUE;
parser.minLenFraction=0f;
parser.requireBothBad=false;
parser.maxNs=-1;
boolean ordered_=false;
int restrictLeft_=0, restrictRight_=0, speed_=0, qSkip_=1;
int ambigMode_=AMBIG_RANDOM;
int matchMode_=MATCH_ALL;
boolean keepPairsTogether_=true;
boolean printNonZeroOnly_=true;
boolean rename_=false, useRefNames_=false;
boolean ecc_=false;
int clearzone_=0;
float clearzoneFraction_=0;
{//Add a fake first scaffold so that the true first scaffold has ID 1, not 0.
scaffoldNames.add("");
scaffoldLengths.add(0);
scaffoldKmers.add(0);
scaffolds.add(null);
}
/* Parse arguments */
for(int i=0; i<args.length; i++){
final String arg=args[i];
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(Parser.parseZip(arg, a, b)){
//do nothing
}else if(Parser.parseHist(arg, a, b)){
//do nothing
}else if(Parser.parseCommonStatic(arg, a, b)){
//do nothing
}else if(Parser.parseQualityAdjust(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(parser.parseInterleaved(arg, a, b)){
//do nothing
}else if(parser.parseTrim(arg, a, b)){
//do nothing
}else if(parser.parseCardinality(arg, a, b)){
//do nothing
}else if(parser.parseCommon(arg, a, b)){
//do nothing
}else if(a.equals("in") || a.equals("in1")){
in1=b;
}else if(a.equals("in2")){
in2=b;
}else if(a.equals("qfin") || a.equals("qfin1")){
qfin1=b;
}else if(a.equals("qfin2")){
qfin2=b;
}else if(a.equals("out") || a.equals("out1") || a.equals("outm") || a.equals("outm1") || a.equals("outmatched") || a.equals("outmatched1")){
outm1=b;
}else if(a.equals("out2") || a.equals("outm") || a.equals("outm2") || a.equals("outmatched") || a.equals("outmatched2")){
outm2=b;
}else if(a.equals("outu") || a.equals("outu1") || a.equals("outunmatched") || a.equals("outunmatched1")){
outu1=b;
}else if(a.equals("outu2") || a.equals("outunmatched") || a.equals("outunmatched2")){
outu2=b;
}else if(a.equals("outpattern") || a.equals("pattern") || a.equals("basename")){
outpattern=b;
}else if(a.equals("stats") || a.equals("scafstats")){
outstats=b;
}else if(a.equals("refstats")){
outrefstats=b;
}else if(a.equals("rpkm") || a.equals("fpkm") || a.equals("cov") || a.equals("coverage") || a.equals("covstats")){
outrpkm=b;
}else if(a.equals("tax") || a.equals("taxa") || a.equals("outtax")){
outtax=b;
}else if(a.equals("outbarcodes") || a.equals("outbc")){
outbarcodes=b;
}else if(a.equals("ref")){
ref.clear();
String[] b2=(b==null) ? null : (new File(b).exists() ? new String[] {b} : b.split(","));
for(String b3 : b2){ref.add(b3);}
}else if(a.equals("literal")){
literal=(b==null) ? null : b.split(",");
// assert(false) : b+", "+Arrays.toString(literal);
}else if(a.equals("forest")){
boolean x=Parse.parseBoolean(b);
if(x){tableType_=AbstractKmerTable.FOREST2D;}
}else if(a.equals("array") || a.equals("array2")){
boolean x=Parse.parseBoolean(b);
if(x){tableType_=AbstractKmerTable.ARRAY2D;}
}else if(a.equals("array1")){
boolean x=Parse.parseBoolean(b);
if(x){tableType_=AbstractKmerTable.ARRAY1D;}
}else if(a.equals("arrayh") || a.equals("hybrid")){
boolean x=Parse.parseBoolean(b);
if(x){tableType_=AbstractKmerTable.ARRAYH;}
}else if(a.equals("arrayhf") || a.equals("hybridfast")){
boolean x=Parse.parseBoolean(b);
if(x){tableType_=AbstractKmerTable.ARRAYHF;}
}else if(a.equals("ways")){
ways_=Integer.parseInt(b);
}else if(a.equals("ordered") || a.equals("ord")){
ordered_=Parse.parseBoolean(b);
outstream.println("Set ORDERED to "+ordered_);
}else if(a.equals("k")){
assert(b!=null) : "\nThe k key needs an integer value greater than 0, such as k=27\n";
k_=Integer.parseInt(b);
assert(k_>0 && k_<32) : "k must be at least 1; default is 31.";
}else if(a.equals("hdist") || a.equals("hammingdistance")){
hammingDistance=Integer.parseInt(b);
assert(hammingDistance>=0 && hammingDistance<4) : "hamming distance must be between 0 and 3; default is 0.";
}else if(a.equals("qhdist") || a.equals("queryhammingdistance")){
qHammingDistance=Integer.parseInt(b);
assert(qHammingDistance>=0 && qHammingDistance<4) : "hamming distance must be between 0 and 3; default is 0.";
}else if(a.equals("edits") || a.equals("edist") || a.equals("editdistance")){
editDistance=Integer.parseInt(b);
assert(editDistance>=0 && editDistance<=3) : "edit distance must be between 0 and 2; default is 0.";
}else if(a.equals("skip") || a.equals("refskip") || a.equals("rskip")){
refSkip=Integer.parseInt(b);
}else if(a.equals("qskip")){
qSkip_=Integer.parseInt(b);
}else if(a.equals("speed")){
speed_=Integer.parseInt(b);
assert(speed_>=0 && speed_<=16) : "Speed range is 0 to 16. Value: "+speed_;
}else if(a.equals("skipreads")){
skipreads_=Parse.parseKMG(b);
}else if(a.equals("minkmerhits") || a.equals("minhits") || a.equals("mh") || a.equals("mkh")){
minKmerHits_=Integer.parseInt(b);
}else if(a.equals("minkmerfraction") || a.equals("minfraction") || a.equals("mkf")){
minKmerFraction_=Float.parseFloat(b);
}else if(a.equals("showspeed") || a.equals("ss")){
showSpeed=Parse.parseBoolean(b);
}else if(a.equals("verbose")){
assert(false) : "Verbose flag is currently static final; must be recompiled to change.";
assert(WAYS>1) : "WAYS=1 is for debug mode.";
}else if(a.equals("mm") || a.equals("maskmiddle")){
if(b==null || Tools.startsWithLetter(b)) {
maskMiddle=Parse.parseBoolean(b);
}else{
midMaskLen=Integer.parseInt(b);
maskMiddle=midMaskLen>0;
}
}else if(a.equals("rcomp")){
rcomp_=Parse.parseBoolean(b);
}else if(a.equals("forbidns") || a.equals("forbidn") || a.equals("fn")){
forbidNs_=Parse.parseBoolean(b);
}else if(a.equals("prealloc") || a.equals("preallocate")){
if(b==null || b.length()<1 || Character.isLetter(b.charAt(0))){
prealloc_=Parse.parseBoolean(b);
}else{
preallocFraction=Tools.max(0, Double.parseDouble(b));
prealloc_=(preallocFraction>0);
}
}else if(a.equals("restrictleft")){
restrictLeft_=Integer.parseInt(b);
}else if(a.equals("restrictright")){
restrictRight_=Integer.parseInt(b);
}else if(a.equals("statscolumns") || a.equals("columns") || a.equals("cols")){
STATS_COLUMNS=Integer.parseInt(b);
assert(STATS_COLUMNS==3 || STATS_COLUMNS==5) : "statscolumns bust be either 3 or 5. Invalid value: "+STATS_COLUMNS;
}else if(a.equals("ambiguous") || a.equals("ambig")){
if(b==null){
throw new RuntimeException(arg);
}else if(b.equalsIgnoreCase("keep") || b.equalsIgnoreCase("best") || b.equalsIgnoreCase("first")){
ambigMode_=AMBIG_FIRST;
}else if(b.equalsIgnoreCase("all")){
ambigMode_=AMBIG_ALL;
}else if(b.equalsIgnoreCase("random") || b.equalsIgnoreCase("rand")){
ambigMode_=AMBIG_RANDOM;
}else if(b.equalsIgnoreCase("toss") || b.equalsIgnoreCase("discard") || b.equalsIgnoreCase("remove")){
ambigMode_=AMBIG_TOSS;
}else{
throw new RuntimeException(arg);
}
}else if(a.equals("match") || a.equals("mode")){
if(b==null){
throw new RuntimeException(arg);
}else if(b.equalsIgnoreCase("all") || b.equalsIgnoreCase("best")){
matchMode_=MATCH_ALL;
}else if(b.equalsIgnoreCase("first")){
matchMode_=MATCH_FIRST;
}else if(b.equalsIgnoreCase("unique") || b.equalsIgnoreCase("firstunique")){
matchMode_=MATCH_UNIQUE;
}else{
throw new RuntimeException(arg);
}
}else if(a.equals("findbestmatch") || a.equals("fbm")){
matchMode_=(Parse.parseBoolean(b) ? MATCH_ALL : MATCH_FIRST);
}else if(a.equals("firstuniquematch") || a.equals("fum")){
if(Parse.parseBoolean(b)){matchMode_=MATCH_UNIQUE;}
}else if(a.equals("keeppairstogether") || a.equals("kpt")){
keepPairsTogether_=Parse.parseBoolean(b);
}else if(a.equals("nzo") || a.equals("nonzeroonly")){
printNonZeroOnly_=Parse.parseBoolean(b);
}else if(a.equals("rename")){
rename_=Parse.parseBoolean(b);
}else if(a.equals("renameduplicates")){
RENAME_DUPLICATES=Parse.parseBoolean(b);
}else if(a.equals("genesets") || a.equals("makesets") || a.equals("makegenesets")){
MAKE_GENE_SETS=Parse.parseBoolean(b);
}else if(a.equals("refnames") || a.equals("userefnames")){
useRefNames_=Parse.parseBoolean(b);
}else if(a.equals("addcount")){
addCount=Parse.parseBoolean(b);
}else if(a.equals("tophitonly")){
topHitOnly=Parse.parseBoolean(b);
}else if(a.equals("addunknown")){
addUnknown=Parse.parseBoolean(b);
}else if(a.equals("initialsize")){
initialSize=Parse.parseIntKMG(b);
}else if(a.equals("dump")){
dump=b;
}else if(a.equals("countvector")){
useCountvector_=Parse.parseBoolean(b);
}else if(a.equals("ecco") || a.equals("ecc")){
ecc_=Parse.parseBoolean(b);
}else if(a.equals("copyundefined") || a.equals("cu")){
REPLICATE_AMBIGUOUS=Parse.parseBoolean(b);
}else if(a.equals("bbsplit")){
BBSPLIT_STYLE=Parse.parseBoolean(b);
}else if(a.equals("table") || a.equals("gi") || a.equals("gitable")){
giTableFile=b;
if("auto".equalsIgnoreCase(b)){giTableFile=TaxTree.defaultTableFile();}
}else if(a.equals("taxnames") || a.equals("taxname")){
taxNameFile=b;
}else if(a.equals("taxnodes") || a.equals("taxnode")){
taxNodeFile=b;
}else if(a.equals("taxtree") || a.equals("tree")){
taxTreeFile=b;
}else if(a.equals("mincount")){
taxNodeCountLimit=Long.parseLong(b);
}else if(a.equals("maxnodes")){
taxNodeNumberLimit=Integer.parseInt(b);
}else if(a.equals("minlevel")){
taxNodeMinLevel=TaxTree.parseLevel(b);
}else if(a.equals("maxlevel")){
taxNodeMaxLevel=TaxTree.parseLevel(b);
}else if(a.equals("clearzone") || a.equals("cz")){
if(b.indexOf('.')>=0) {
clearzoneFraction_=Float.parseFloat(b);
}else {
clearzone_=Integer.parseInt(b);
}
}else if(a.equals("clearzonefraction") || a.equals("czf")){
clearzoneFraction_=Float.parseFloat(b);
}
else if(a.equals("processcontainedref")){
processContainedRef=Parse.parseBoolean(b);
}else if(a.equals("storerefbases")){
storeRefBases=Parse.parseBoolean(b);
}
else if(a.equals("trackbarcodes")){
trackBarcodes=Parse.parseBoolean(b);
}
else if(b==null && new File(arg).exists()){
ref.add(arg);
}else{
throw new RuntimeException("Unknown parameter "+args[i]);
}
}
if("auto".equals(taxTreeFile)){taxTreeFile=TaxTree.defaultTreeFile();}
if(ref.isEmpty()){ref=null;}
{//Process parser fields
Parser.processQuality();
maxReads=parser.maxReads;
samplerate=parser.samplerate;
sampleseed=parser.sampleseed;
overwrite=ReadStats.overwrite=parser.overwrite;
append=ReadStats.append=parser.append;
forceTrimModulo=parser.forceTrimModulo;
forceTrimLeft=parser.forceTrimLeft;
forceTrimRight=parser.forceTrimRight;
forceTrimRight2=parser.forceTrimRight2;
qtrimLeft=parser.qtrimLeft;
qtrimRight=parser.qtrimRight;
trimq=parser.trimq;
trimE=parser.trimE();
minLenFraction=parser.minLenFraction;
minAvgQuality=parser.minAvgQuality;
minAvgQualityBases=parser.minAvgQualityBases;
chastityFilter=parser.chastityFilter;
minReadLength=parser.minReadLength;
maxReadLength=parser.maxReadLength;
maxNs=parser.maxNs;
minConsecutiveBases=parser.minConsecutiveBases;
// minGC=parser.minGC;
// maxGC=parser.maxGC;
// filterGC=parser.filterGC;
// minTrimLength=(parser.minTrimLength>=0 ? parser.minTrimLength : minTrimLength);
// requireBothBad=parser.requireBothBad;
removePairsIfEitherBad=!parser.requireBothBad;
loglog=(parser.loglog ? CardinalityTracker.makeTracker(parser) : null);
loglogOut=(parser.loglogOut ? CardinalityTracker.makeTracker(parser) : null);
THREADS=Shared.threads();
}
refNames.add(null);//Fake first reference also
if(ref!=null){
ArrayList<String> temp=new ArrayList<String>();
for(String s : ref){
if(s==null){
assert(false) : "Null reference file.";
}else if(new File(s).exists()){
Tools.getFileOrFiles(s, temp, true, false, false, false);
}else{
String fname=null;
if("phix".equalsIgnoreCase(s)){
fname=Data.findPath("?phix2.fa.gz");
}else if("polya".equalsIgnoreCase(s) || "polyt".equalsIgnoreCase(s)){
fname=Data.findPath("?polyA.fa.gz");
}else if("lambda".equalsIgnoreCase(s)){
fname=Data.findPath("?lambda.fa.gz");
}else if("kapa".equalsIgnoreCase(s)){
fname=Data.findPath("?kapatags.L40.fa");
}else if("pjet".equalsIgnoreCase(s)){
fname=Data.findPath("?pJET1.2.fa");
}else if("mtst".equalsIgnoreCase(s)){
fname=Data.findPath("?mtst.fa");
}else if("adapters".equalsIgnoreCase(s)){
fname=Data.findPath("?adapters.fa");
}else if("truseq".equalsIgnoreCase(s)){
fname=Data.findPath("?truseq.fa.gz");
}else if("nextera".equalsIgnoreCase(s)){
fname=Data.findPath("?nextera.fa.gz");
}else if("artifacts".equalsIgnoreCase(s)){
fname=Data.findPath("?sequencing_artifacts.fa.gz");
}else if("crisprs".equalsIgnoreCase(s)){
s=Data.findPath("?crisprs.fa.gz");
}else{
assert(false) : "Can't find reference file "+s;
}
temp.add(fname);
}
}
ref=temp;
if(ref.size()<1){ref=null;}
refNames.addAll(temp);
}
// if(ref!=null){
// ArrayList<String> temp=new ArrayList<String>();
// for(String s : ref){
// Tools.getFileOrFiles(s, temp, true, false, false, false);
// }
// ref=temp.toArray(new String[0]);
// if(ref.length<1){ref=null;}
// refNames.addAll(temp);
// }
if(literal!=null){refNames.add("literal");}
refScafCounts=new int[refNames.size()];
if(prealloc_){
outstream.println("Note - if this program runs out of memory, please disable the prealloc flag.");
}
/* Set final variables; post-process and validate argument combinations */
tableType=tableType_;
hammingDistance=Tools.max(editDistance, hammingDistance);
refSkip=Tools.max(0, refSkip);
rcomp=rcomp_;
forbidNs=(forbidNs_ || hammingDistance<1);
skipreads=skipreads_;
ordered=ordered_;
restrictLeft=Tools.max(restrictLeft_, 0);
restrictRight=Tools.max(restrictRight_, 0);
ambigMode=ambigMode_;
matchMode=matchMode_;
keepPairsTogether=keepPairsTogether_;
printNonZeroOnly=printNonZeroOnly_;
rename=rename_;
useRefNames=useRefNames_;
speed=speed_;
qSkip=qSkip_;
noAccel=(speed<1 && qSkip<2);
clearzone=clearzone_;
clearzoneFraction=clearzoneFraction_;
parsecustom=FASTQ.PARSE_CUSTOM;
ecc=ecc_;
USE_TAXTREE=(taxNameFile!=null || taxNodeFile!=null || outtax!=null || taxTreeFile!=null);
USE_COUNTVECTOR=useCountvector_;
if(outbarcodes!=null){trackBarcodes=true;}
if(trackBarcodes){
barcodeData=new BarcodeMappingStats();
}
if((speed>0 && qSkip>1) || (qSkip>1 && refSkip>1) || (speed>0 && refSkip>1)){
outstream.println("WARNING: It is not recommended to use more than one of qskip, speed, and rskip together.");
outstream.println("qskip="+qSkip+", speed="+speed+", rskip="+refSkip);
}
{
long usableMemory;
long tableMemory;
{
long memory=Runtime.getRuntime().maxMemory();
double xmsRatio=Shared.xmsRatio();
usableMemory=(long)Tools.max(((memory-96000000-(20*400000 /* for atomic arrays */))*(xmsRatio>0.97 ? 0.82 : 0.72)), memory*0.45);
tableMemory=(long)(usableMemory*.95);
}
if(initialSize<1){
final int factor=(tableType==AbstractKmerTable.ARRAY1D ? 12 : tableType==AbstractKmerTable.ARRAYH ? 22 : tableType==AbstractKmerTable.ARRAYHF ? 22 : 27);
final long memOverWays=tableMemory/(factor*WAYS);
final double mem2=(prealloc_ ? preallocFraction : 1)*tableMemory;
initialSize=(prealloc_ || memOverWays<initialSizeDefault ? (int)Tools.min(2142000000, (long)(mem2/(factor*WAYS))) : initialSizeDefault);
if(initialSize!=initialSizeDefault){
outstream.println("Initial size set to "+initialSize);
}
}
}
k=k_;
k2=k-1;
if(maskMiddle) {
midMaskLen=(midMaskLen>0 ? midMaskLen : 2-(k&1));
}else{
midMaskLen=0;
}
minKmerHits=minKmerHits_;
minKmerFraction=Tools.max(minKmerFraction_, 0);
assert(minKmerHits>=1) : "minKmerHits must be at least 1; value="+minKmerHits;
assert(minKmerFraction<=1) : "minKmerFraction must range from 0 to 1; value="+minKmerFraction;
kfilter=(ref!=null || literal!=null);
assert(kfilter==false || (k>0 && k<32)) : "K must range from 1 to 31.";
if(maskMiddle){
final int bitsPerBase=2;
assert(k>midMaskLen+1);
int bits=midMaskLen*bitsPerBase;
// int shift=(k-maskMiddle)&(~1);//Equivalent to (x/2)*2
int shift=((k-midMaskLen)/2)*bitsPerBase; //old behavior before moving to variable width can be restored with +1: "((k-maskMiddle+1)/2)*bitsPerBase"
middleMask=~((~((-1L)<<bits))<<shift);
// mask<<=(shift);
}else{
middleMask=-1L;
}
// middleMask=maskMiddle ? ~(3L<<(2*(k/2))) : -1L;
/* Adjust I/O settings and filenames */
assert(FastaReadInputStream.settingsOK());
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
if(in1!=null && in1.contains("#") && !new File(in1).exists()){
int pound=in1.lastIndexOf('#');
String a=in1.substring(0, pound);
String b=in1.substring(pound+1);
in1=a+1+b;
in2=a+2+b;
}
if(in2!=null && (in2.contains("=") || in2.equalsIgnoreCase("null"))){in2=null;}
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;
}
if(qfin1!=null && qfin1.contains("#") && in2!=null && !new File(qfin1).exists()){
int pound=qfin1.lastIndexOf('#');
String a=qfin1.substring(0, pound);
String b=qfin1.substring(pound+1);
qfin1=a+1+b;
qfin2=a+2+b;
}
if(outu1!=null && outu1.contains("#")){
int pound=outu1.lastIndexOf('#');
String a=outu1.substring(0, pound);
String b=outu1.substring(pound+1);
outu1=a+1+b;
outu2=a+2+b;
}
if(outm1!=null && outm1.contains("#")){
int pound=outm1.lastIndexOf('#');
String a=outm1.substring(0, pound);
String b=outm1.substring(pound+1);
outm1=a+1+b;
outm2=a+2+b;
}
if((outu2!=null || outm2!=null) && (in1!=null && in2==null)){
if(!FASTQ.FORCE_INTERLEAVED){outstream.println("Forcing interleaved input because paired output was specified for a single input file.");}
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=true;
}
if(!Tools.testOutputFiles(overwrite, append, false, outu1, outu2, outm1, outm2, outpattern, outstats, outrpkm, outrefstats)){
throw new RuntimeException("\nCan't write to some output files; overwrite="+overwrite+"\n");
}
if(!Tools.testInputFiles(false, true, in1, in2, qfin1, qfin2, taxNameFile, taxNodeFile, giTableFile, taxTreeFile)){
throw new RuntimeException("\nCan't read some input files.\n");
}
if(!Tools.testInputFiles(true, true, ref)){
throw new RuntimeException("\nCan't read to some reference files.\n");
}
if(!Tools.testForDuplicateFiles(true, in1, in2, qfin1, qfin2, outu1, outu2, outm1, outm2, outpattern, outstats, outrpkm, outrefstats)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
assert(THREADS>0) : "THREADS must be greater than 0.";
assert(in1==null || in1.toLowerCase().startsWith("stdin") || in1.toLowerCase().startsWith("standardin") || new File(in1).exists()) : "Can't find "+in1;
assert(in2==null || in2.toLowerCase().startsWith("stdin") || in2.toLowerCase().startsWith("standardin") || new File(in2).exists()) : "Can't find "+in2;
if(ref==null && literal==null){
outstream.println("ERROR: No reference sequences specified. Use the -da flag to run anyway.");
assert(false) : "Please specify a reference.";
}
if(ref!=null){
for(String s0 : ref){
assert(s0!=null) : "Specified a null reference.";
String s=s0.toLowerCase();
assert(s==null || s.startsWith("stdin") || s.startsWith("standardin") || new File(s0).exists()) : "Can't find "+s0;
}
}
if(MAKE_GENE_SETS) {
setMap=new ConcurrentHashMap<String, StringCount>();
}
//Initialize tables
ScheduleMaker scheduleMaker=new ScheduleMaker(WAYS, 14, prealloc_, (prealloc_ ? preallocFraction : 0.9));
int[] schedule=scheduleMaker.makeSchedule();
keySets=AbstractKmerTable.preallocate(WAYS, tableType, schedule, -1L);
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* Primary driver function of class.
*/
public void process(){
/* Check for output file collisions */
if(!Tools.testOutputFiles(overwrite, append, false, outu1, outu2, outm1, outm2, outstats, outrpkm, outrefstats)){
throw new RuntimeException("One or more output files were duplicate or could not be written to. Check the names or set the 'overwrite=true' flag.");
}
/* Start overall timer */
Timer t=new Timer();
// boolean dq0=FASTQ.DETECT_QUALITY;
// boolean ti0=FASTQ.TEST_INTERLEAVED;
// int rbl0=Shared.bufferLen();;
// FASTQ.DETECT_QUALITY=false;
// FASTQ.TEST_INTERLEAVED=false;
// Shared.setBufferLen(16;
process2(t.time1);
// FASTQ.DETECT_QUALITY=dq0;
// FASTQ.TEST_INTERLEAVED=ti0;
// Shared.setBufferLen(rbl0;
/* Stop timer and calculate speed statistics */
t.stop();
lastReadsOut=readsUnmatched;
if(showSpeed){
outstream.println();
outstream.println(Tools.timeReadsBasesProcessed(t, readsIn, basesIn, 8));
}
if(outstream!=System.err && outstream!=System.out){outstream.close();}
/* Throw an exception if errors were detected */
if(errorState){
throw new RuntimeException("Seal terminated in an error state; the output may be corrupt.");
}
}
public void process2(long startTime){
/* Start phase timer */
Timer t=new Timer();
if(DISPLAY_PROGRESS){
outstream.println("Initial:");
Shared.printMemory();
outstream.println();
}
/* Fill tables with reference kmers */
{
final boolean oldTI=FASTQ.TEST_INTERLEAVED; //TODO: This needs to be changed to a non-static field, or somehow 'read mode' and 'ref mode' need to be distinguished.
final boolean oldFI=FASTQ.FORCE_INTERLEAVED;
final boolean oldSplit=FastaReadInputStream.SPLIT_READS;
final int oldML=FastaReadInputStream.MIN_READ_LEN;
FASTQ.TEST_INTERLEAVED=false;
FASTQ.FORCE_INTERLEAVED=false;
FastaReadInputStream.SPLIT_READS=false;
FastaReadInputStream.MIN_READ_LEN=1;
storedKmers=spawnLoadThreads();
FASTQ.TEST_INTERLEAVED=oldTI;
FASTQ.FORCE_INTERLEAVED=oldFI;
FastaReadInputStream.SPLIT_READS=oldSplit;
FastaReadInputStream.MIN_READ_LEN=oldML;
// if(useRefNames){toRefNames();}
t.stop();
}
/* Check memory */
{
long ram=freeMemory();
ALLOW_LOCAL_ARRAYS=(scaffoldNames!=null && Tools.max(THREADS, 1)*3*8*scaffoldNames.size()<ram*5);
}
/* Dump kmers to text */
if(dump!=null){
ByteStreamWriter bsw=new ByteStreamWriter(dump, overwrite, false, true);
bsw.start();
for(AbstractKmerTable set : keySets){
set.dumpKmersAsBytes(bsw, k, 0, Integer.MAX_VALUE, null);
}
bsw.poisonAndWait();
}
final boolean vic=Read.VALIDATE_IN_CONSTRUCTOR;
Read.VALIDATE_IN_CONSTRUCTOR=THREADS<4;
/* Do kmer matching of input reads */
spawnProcessThreads(t);
Read.VALIDATE_IN_CONSTRUCTOR=vic;
/* Unload kmers to save memory */
if(RELEASE_TABLES){
unloadKmers();
}
if(USE_TAXTREE){
if(giTableFile!=null){loadGiToTaxid();}
if(USE_TAXTREE){tree=TaxTree.loadTaxTree(taxTreeFile, taxNameFile, taxNodeFile, null, DISPLAY_PROGRESS ? outstream : null, false, false);}
addToTree();
}
/* Write statistics to files */
writeStats();
writeRPKM();
writeBarcodeStats();
addToRqcMap();
if(!BBSPLIT_STYLE){
writeRefStats();
}else{
writeRefStats_BBSplitStyle(readsIn);
}
writeTaxonomy();
/* Unload sequence data to save memory */
if(RELEASE_TABLES){
unloadScaffolds();
tree=null;
GiToTaxid.unload();
}
outstream.println("\nInput: \t"+readsIn+" reads \t\t"+basesIn+" bases.");
if(ref!=null || literal!=null){
outstream.println("Matched reads: \t"+readsMatched+" reads ("+toPercent(readsMatched, readsIn)+") \t"+
basesMatched+" bases ("+toPercent(basesMatched, basesIn)+")");
outstream.println("Unmatched reads: \t"+readsUnmatched+" reads ("+toPercent(readsUnmatched, readsIn)+") \t"+
basesUnmatched+" bases ("+toPercent(basesUnmatched, basesIn)+")");
outstream.flush();
}
if(qtrimLeft || qtrimRight){
outstream.println("QTrimmed: \t"+readsQTrimmed+" reads ("+toPercent(readsQTrimmed, readsIn)+") \t"+
basesQTrimmed+" bases ("+toPercent(basesQTrimmed, basesIn)+")");
}
if(forceTrimLeft>0 || forceTrimRight>0 || forceTrimRight2>0 || forceTrimModulo>0){
outstream.println("FTrimmed: \t"+readsFTrimmed+" reads ("+toPercent(readsFTrimmed, readsIn)+") \t"+
basesFTrimmed+" bases ("+toPercent(basesFTrimmed, basesIn)+")");
}
if(minAvgQuality>0 || maxNs>=0){
outstream.println("Low quality discards: \t"+readsQFiltered+" reads ("+toPercent(readsQFiltered, readsIn)+") \t"+
basesQFiltered+" bases ("+toPercent(basesQFiltered, basesIn)+")");
}
if(loglog!=null){
outstream.println("Unique "+loglog.k+"-mers: \t"+loglog.cardinality());
}
if(loglogOut!=null){
outstream.println("Unique "+loglogOut.k+"-mers out: \t"+loglogOut.cardinality());
}
if(parsecustom){
outstream.println();
outstream.println("Correctly mapped: \t"+correctReads+" reads ("+toPercent(correctReads, readsIn)+")");
outstream.println("Incorrectly mapped: \t"+incorrectReads+" reads ("+toPercent(incorrectReads, readsIn)+")");
}
// outstream.println("Result: \t"+readsMatched+" reads ("+toPercent(readsMatched*100.0/readsIn)+"%) \t"+
// basesMatched+" bases ("+toPercent(basesMatched*100.0/basesIn)+"%)");
}
private static String toPercent(long numerator, long denominator){
if(denominator<1){return "0.00%";}
return Tools.format("%.2f%%",numerator*100.0/denominator);
}
/**
* Clear stored kmers.
*/
public void unloadKmers(){
if(keySets!=null){
for(int i=0; i<keySets.length; i++){keySets[i]=null;}
}
}
/**
* Clear stored sequence data.
*/
public void unloadScaffolds(){
if(scaffoldNames!=null && !scaffoldNames.isEmpty()){
scaffoldNames.clear();
scaffoldNames.trimToSize();
}
scaffoldReadCounts=null;
scaffoldFragCounts=null;
scaffoldBaseCounts=null;
scaffoldAmbigReadCounts=null;
scaffoldLengths=null;
scaffoldKmers=null;
scaffolds=null;
}
private void addToRqcMap(){
BBDuk.putRqc("inputReads", readsIn, false, false);
BBDuk.putRqc("inputBases", basesIn, false, false);
if(qtrimLeft || qtrimRight){
BBDuk.putRqc("qtrimmedReads", readsQTrimmed, false, true);
BBDuk.putRqc("qtrimmedBases", basesQTrimmed, false, true);
}
BBDuk.putRqc("qfilteredReads", readsQFiltered, false, true);
BBDuk.putRqc("qfilteredBases", basesQFiltered, false, true);
{//This is kind of a hack, to match BBDuk's syntax for RQCFilter.
BBDuk.putRqc("kfilteredReads", readsMatched, false, true);
BBDuk.putRqc("kfilteredBases", basesMatched, false, true);
BBDuk.putRqc("outputReads", readsUnmatched, true, false);
BBDuk.putRqc("outputBases", basesUnmatched, true, false);
}
}
/**
* Write statistics about how many reads matched each reference scaffold.
*/
private void writeStats(){
if(outstats==null){return;}
final TextStreamWriter tsw=new TextStreamWriter(outstats, overwrite, false, false);
tsw.start();
long rsum=0, bsum=0, asum=0;
/* Create StringCount list of scaffold names and hitcounts */
ArrayList<StringCount> list=new ArrayList<StringCount>();
for(int i=1; i<scaffoldNames.size(); i++){
final long num1=scaffoldReadCounts.get(i), num2=scaffoldBaseCounts.get(i), num3=scaffoldAmbigReadCounts.get(i);
if(num1>0 || !printNonZeroOnly){
rsum+=num1;
bsum+=num2;
asum+=num3;
final String s=scaffoldNames.get(i);
final int len=scaffoldLengths.get(i);
final StringCount sn=new StringCount(s, len, num1, num2, num3);
list.add(sn);
}
}
Shared.sort(list);
if(MAKE_GENE_SETS) {
ArrayList<StringCount> list2=new ArrayList<StringCount>(setMap.values());
Shared.sort(list2);
list.addAll(list2);
}
final double rmult=100.0/(readsIn>0 ? readsIn : 1);
final double bmult=100.0/(basesIn>0 ? basesIn : 1);
tsw.print("#File\t"+in1+(in2==null ? "" : "\t"+in2)+"\n");
if(STATS_COLUMNS==3){
tsw.print(Tools.format("#Total\t%d\n",readsIn));
// tsw.print(Tools.format("#Matched\t%d\t%.5f%%\n",rsum,rmult*rsum)); //With ambig=all, gives over 100%
tsw.print(Tools.format("#Matched\t%d\t%.5f%%\n",readsMatched,rmult*readsMatched));
tsw.print("#Name\tReads\tReadsPct\n");
for(int i=0; i<list.size(); i++){
StringCount sn=list.get(i);
tsw.print(Tools.format("%s\t%d\t%.5f%%\n",sn.name,sn.reads,(sn.reads*rmult)));
}
}else{
tsw.print(Tools.format("#Total\t%d\t%d\n",readsIn,basesIn));
// tsw.print(Tools.format("#Matched\t%d\t%.5f%%\n",rsum,rmult*rsum,bsum,bsum*bmult)); //With ambig=all, gives over 100%
tsw.print(Tools.format("#Matched\t%d\t%.5f%%\t%d\n",readsMatched,rmult*readsMatched,basesMatched,basesMatched*bmult,asum));
tsw.print("#Name\tReads\tReadsPct\tBases\tBasesPct\tAmbigReads\n");
for(int i=0; i<list.size(); i++){
StringCount sn=list.get(i);
tsw.print(Tools.format("%s\t%d\t%.5f%%\t%d\t%.5f%%\t%d\n",sn.name,sn.reads,(sn.reads*rmult),sn.bases,(sn.bases*bmult),sn.ambigReads));
}
}
tsw.poisonAndWait();
}
private void writeBarcodeStats(){
if(barcodeData==null || outbarcodes==null) {return;}
barcodeData.writeStats(outbarcodes, overwrite);
}
private void writeRPKM(){
writeRPKM(outrpkm, in1, in2, readsIn, printNonZeroOnly,
scaffoldNames, scaffoldLengths,
scaffoldReadCounts, scaffoldFragCounts, scaffoldBaseCounts, scaffoldAmbigReadCounts);
}
/**
* Write RPKM statistics.
*/
public void writeRPKM(String out, String in1, String in2, long readsIn, boolean printNonZeroOnly,
ArrayList<String> scaffoldNames, IntList scaffoldLengths, AtomicLongArray scaffoldReadCounts,
AtomicLongArray scaffoldFragCounts, AtomicLongArray scaffoldBaseCounts, AtomicLongArray scaffoldAmbigReadCounts){
if(out==null){return;}
final TextStreamWriter tsw=new TextStreamWriter(out, overwrite, false, false);
tsw.start();
/* Count mapped reads */
long mappedReads=0;
long mappedFrags=0;
for(int i=0; i<scaffoldReadCounts.length(); i++){
mappedReads+=scaffoldReadCounts.get(i);
mappedFrags+=scaffoldFragCounts.get(i);
}
/* Print header */
tsw.print("#File\t"+in1+(in2==null ? "" : "\t"+in2)+"\n");
tsw.print(Tools.format("#Reads\t%d\n",readsIn));
// tsw.print(Tools.format("#Mapped\t%d\n",mappedReads));
tsw.print(Tools.format("#Mapped\t%d\n",readsMatched));
tsw.print(Tools.format("#RefSequences\t%d\n",Tools.max(0, scaffoldNames.size()-1)));
tsw.print("#Name\tLength\tBases\tCoverage\tReads\tRPKM\tFrags\tFPKM\tAmbigReads\n");
final float readMult=1000000000f/Tools.max(1, mappedReads);
final float fragMult=1000000000f/Tools.max(1, mappedFrags);
/* Print data */
for(int i=1; i<scaffoldNames.size(); i++){
final long reads=scaffoldReadCounts.get(i);
final long frags=scaffoldFragCounts.get(i);
final long bases=scaffoldBaseCounts.get(i);
final long ambig=scaffoldAmbigReadCounts.get(i);
final String s=scaffoldNames.get(i);
final int len=scaffoldLengths.get(i);
final double invlen=1.0/Tools.max(1, len);
final double readMult2=readMult*invlen;
final double fragMult2=fragMult*invlen;
if(reads>0 || !printNonZeroOnly){
tsw.print(Tools.format("%s\t%d\t%d\t%.4f\t%d\t%.4f\t%d\t%.4f\t%d\n",s,len,bases,bases*invlen,reads,reads*readMult2,frags,frags*fragMult2,ambig));
}
}
if(MAKE_GENE_SETS) {
ArrayList<StringCount> list=new ArrayList<StringCount>(setMap.values());
Shared.sort(list);
for(StringCount sc : list) {
String s=sc.name;
final long reads=sc.reads;
final long frags=sc.reads;
final long bases=sc.bases;
final long ambig=sc.ambigReads;
final int len=sc.length;
final double invlen=1.0/Tools.max(1, len);
final double readMult2=readMult*invlen;
final double fragMult2=fragMult*invlen;
if(reads>0 || !printNonZeroOnly){
tsw.print(Tools.format("%s\t%d\t%d\t%.4f\t%d\t%.4f\t%d\t%.4f\t%d\n",s,len,bases,bases*invlen,reads,reads*readMult2,frags,frags*fragMult2,ambig));
}
}
}
tsw.poisonAndWait();
}
/**
* Write statistics on a per-reference basis.
*/
private void writeRefStats(){
if(outrefstats==null){return;}
final TextStreamWriter tsw=new TextStreamWriter(outrefstats, overwrite, false, false);
tsw.start();
/* Count mapped reads */
long mapped=0;
for(int i=0; i<scaffoldReadCounts.length(); i++){
mapped+=scaffoldReadCounts.get(i);
}
final int numRefs=refNames.size();
final int totalScafs=scaffoldReadCounts.length();
final int totalScafs2=(int)shared.Vector.sum(refScafCounts);
assert(totalScafs<=totalScafs2+1) : totalScafs+", "+totalScafs2;
long[] refReadCounts=new long[numRefs];
long[] refFragCounts=new long[numRefs];
long[] refBaseCounts=new long[numRefs];
long[] refLengths=new long[numRefs];
long[] ambigReadCounts=new long[numRefs];
for(int refNum=1, scafNum=1; refNum<numRefs; refNum++){
final int scafs=(useRefNames ? 1 : refScafCounts[refNum]);
final int scafLimitThisRef=scafNum+scafs;
// System.err.println("ref="+refNum+", scafs="+scafs+", scafnum="+scafNum+", limit="+scafLimitThisRef+", max="+totalScafs);
assert(refScafCounts[refNum]>0) : refNum;
assert(scafLimitThisRef<=scaffoldReadCounts.length()) : scafLimitThisRef+", "+scaffoldReadCounts.length()+", "+refNum+", "+refScafCounts[refNum]+", "+Arrays.toString(refScafCounts);
while(scafNum<scafLimitThisRef){
refReadCounts[refNum]+=scaffoldReadCounts.get(scafNum);
refFragCounts[refNum]+=scaffoldFragCounts.get(scafNum);
refBaseCounts[refNum]+=scaffoldBaseCounts.get(scafNum);
refLengths[refNum]+=scaffoldLengths.get(scafNum);
ambigReadCounts[refNum]+=scaffoldAmbigReadCounts.get(scafNum);
scafNum++;
}
}
/* Print header */
tsw.print("#File\t"+in1+(in2==null ? "" : "\t"+in2)+"\n");
tsw.print(Tools.format("#Reads\t%d\n",readsIn));
tsw.print(Tools.format("#Mapped\t%d\n",mapped));
tsw.print(Tools.format("#References\t%d\n",refNames.size()-1));
tsw.print("#Name\tLength\tScaffolds\tBases\tCoverage\tReads\tRPKM\tFrags\tFPKM\tAmbigReads\n");
final float mult=1000000000f/Tools.max(1, mapped);
/* Print data */
for(int i=1; i<refNames.size(); i++){
final long reads=refReadCounts[i];
final long frags=refFragCounts[i];
final long bases=refBaseCounts[i];
final long len=refLengths[i];
final long ambig=ambigReadCounts[i];
final int scafs=refScafCounts[i];
final String name=ReadWrite.stripToCore(refNames.get(i));
final double invlen=1.0/Tools.max(1, len);
final double mult2=mult*invlen;
if(reads>0 || !printNonZeroOnly){
tsw.print(Tools.format("%s\t%d\t%d\t%d\t%.4f\t%d\t%.4f\t%d\t%.4f\t%d\n",name,len,scafs,bases,bases*invlen,reads,reads*mult2,frags,frags*mult2,ambig));
}
}
tsw.poisonAndWait();
}
/**
* Write statistics on a per-reference basis.
*/
private void writeRefStats_BBSplitStyle(long totalReads){
if(outrefstats==null){return;}
final TextStreamWriter tsw=new TextStreamWriter(outrefstats, overwrite, false, false);
tsw.start();
final int numRefs=refNames.size();
long[] refReadCounts=new long[numRefs];
long[] refBaseCounts=new long[numRefs];
long[] refAmbigCounts=new long[numRefs];
for(int r=1, s=1; r<numRefs; r++){
final int lim=s+(useRefNames ? 1 : refScafCounts[r]);
while(s<lim){
refReadCounts[r]+=scaffoldReadCounts.get(s);
refBaseCounts[r]+=scaffoldBaseCounts.get(s);
refAmbigCounts[r]+=scaffoldAmbigReadCounts.get(s);
s++;
}
}
/* Print header */
tsw.print("#name\t%unambiguousReads\tunambiguousMB\t%ambiguousReads\tambiguousMB\tunambiguousReads\tambiguousReads\n");
final float rmult=100f/Tools.max(1, totalReads);
/* Print data */
for(int i=1; i<refNames.size(); i++){
final long reads=refReadCounts[i];
final long bases=refBaseCounts[i];
final float unambigMB=bases*0.000001f;
final long ambigReads=refAmbigCounts[i];
final long ambigBases=0; //TODO but not urgent
final float ambigMB=ambigBases*0.000001f;
final String name=ReadWrite.stripToCore(refNames.get(i));
final double unambigReadP=rmult*reads;
final double ambigReadP=rmult*ambigReads;
if(reads>0 || !printNonZeroOnly){
tsw.print(Tools.format("%s\t%.5f\t%.5f\t%.5f\t%.5f\t%d\t%d\n",name,unambigReadP,unambigMB,ambigReadP,ambigMB,reads,ambigReads));
}
}
tsw.poisonAndWait();
}
/**
* Write taxonomic information.
*/
private void writeTaxonomy(){
if(!USE_TAXTREE || outtax==null){return;}
long mappedFrags=0;
for(int i=0; i<scaffoldReadCounts.length(); i++){
mappedFrags+=scaffoldFragCounts.get(i);
}
final double fragMult=100.0/Tools.max(1, fragsIn);
final TextStreamWriter tsw=new TextStreamWriter(outtax, overwrite, false, false);
tsw.start();
tsw.print("#File\t"+in1+(in2==null ? "" : "\t"+in2)+"\n");
tsw.print(Tools.format("#Reads\t%d\n",fragsIn));
tsw.print(Tools.format("#Mapped\t%d\n",mappedFrags));
tsw.print(Tools.format("#Limits\t%d\t%d\t%d\t%d\n", taxNodeCountLimit, taxNodeNumberLimit, taxNodeMinLevel, taxNodeMaxLevel));
tsw.print("#ID\tCount\tPercent\tLevel\tName\n");
// assert(false) : taxNodeCountLimit+", "+taxNodeMinLevel+", "+taxNodeMaxLevel;
ArrayList<TaxNode> nodes=tree.gatherNodesAtLeastLimit(taxNodeCountLimit, taxNodeMinLevel, taxNodeMaxLevel);
for(int i=0, cap=Tools.min(nodes.size(), (taxNodeNumberLimit>0 ? taxNodeNumberLimit : Integer.MAX_VALUE)); i<cap; i++){
TaxNode n=nodes.get(i);
tsw.print(Tools.format("%d\t%d\t%.4f\t%s\t%s\n", n.id, n.countSum, n.countSum*fragMult, n.levelStringExtended(false), n.name));
}
tsw.poisonAndWait();
}
// /**
// * Fills the scaffold names array with reference names.
// */
// private void toRefNames(){
// final int numRefs=refNames.size();
// for(int r=0, s=1; r<numRefs; r++){
// final int scafs=refScafCounts[r];
// final int lim=s+scafs;
// final String name=ReadWrite.stripToCore(refNames.get(r));
//// outstream.println("r="+r+", s="+s+", scafs="+scafs+", lim="+lim+", name="+name);
// while(s<lim){
//// outstream.println(r+", "+s+". Setting "+scaffoldNames.get(s)+" -> "+name);
// scaffoldNames.set(s, name);
// s++;
// }
// }
// }
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
private static int numKmers(Read r1, Read r2, int k){
int x=0;
if(r1!=null){
x+=Tools.max(r1.length()-k+1, 0);
}
if(r2!=null){
x+=Tools.max(r2.length()-k+1, 0);
}
return x;
}
private void loadGiToTaxid(){
Timer t=new Timer();
outstream.println("Loading gi to taxa translation table.");
GiToTaxid.initialize(giTableFile);
t.stop();
if(DISPLAY_PROGRESS){
outstream.println("Time: \t"+t);
Shared.printMemory();
outstream.println();
}
}
// private TaxTree loadTaxTree(){
// assert(taxTreeFile!=null || (taxNameFile!=null && taxNodeFile!=null)) : "Must specify both taxname and taxnode files.";
// Timer t=new Timer();
// outstream.print("\nLoading tax tree; ");
// final TaxTree tree;
// if(taxTreeFile!=null){
// tree=ReadWrite.read(TaxTree.class, taxTreeFile, true);
// }else{
// tree=new TaxTree(taxNameFile, taxNodeFile);
// }
// t.stop();
// if(DISPLAY_PROGRESS){
// outstream.println("time: \t"+t);
// Shared.printMemory();
// outstream.println();
// }
// return tree;
// }
private void addToTree(){
for(int i=0; i<scaffoldFragCounts.length(); i++){
long count=scaffoldFragCounts.get(i);
if(count>0){
String name=scaffoldNames.get(i);
assert(name.startsWith("ncbi|") || name.startsWith("tid|") || (name.startsWith("gi|") && GiToTaxid.isInitialized())) :
"\nFor taxonomy, all ref names must start with 'gi|' or 'ncbi|' or 'tid|'.\n" +
"If the names start with 'gi', the gi= flag must be set.\n";
int id=GiToTaxid.getID(name);
if(id>-1){
tree.incrementRaw(id, count);
}
}
}
tree.percolateUp();
}
/**
* Fills tables with kmers from references, using multiple LoadThread.
* @return Number of kmers stored.
*/
private long spawnLoadThreads(){
Timer t=new Timer();
if((ref==null || ref.size()<1) && (literal==null || literal.length<1)){return 0;}
long added=0;
final boolean oldParseCustom=FASTQ.PARSE_CUSTOM;
FASTQ.PARSE_CUSTOM=false;
/* Create load threads */
LoadThread[] loaders=new LoadThread[WAYS];
for(int i=0; i<loaders.length; i++){
loaders[i]=new LoadThread(i);
loaders[i].start();
}
boolean warned=false;
/* For each reference file... */
int refNum=1;
if(ref!=null){
HashMap<String, Integer> nameMap=new HashMap<String, Integer>();
for(String refname : ref){
/* Start an input stream */
FileFormat ff=FileFormat.testInput(refname, FileFormat.FASTA, null, true, true);
ConcurrentReadInputStream cris=ConcurrentReadInputStream.getReadInputStream(-1L, false, ff, null, null, null, Shared.USE_MPI, true);
cris.start(); //4567
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
final String core=ReadWrite.stripToCore(refname);
if(useRefNames){
assert(refNum==scaffoldNames.size());
assert(!nameMap.containsKey(core)) : "Duplicate file name: "+core;
Integer id=scaffoldNames.size();
scaffoldNames.add(core);
nameMap.put(core, id);
}
/* Iterate through read lists from the input stream */
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
{
/* Assign a unique ID number to each scaffold */
ArrayList<Read> reads2=new ArrayList<Read>(reads);
for(Read r1 : reads2){
final Read r2=r1.mate;
if(useRefNames){
r1.id=core;
if(r2!=null){r2.id=core;}
}else if(r1.id==null){r1.id=Integer.valueOf(scaffoldNames.size()).toString();}
final Integer id;
{
String name=r1.id;
Integer x=nameMap.get(r1.id);
for(int suffix=2; !useRefNames && x!=null; suffix++){
name=r1.id+"_"+suffix;
if(!warned) {
assert(RENAME_DUPLICATES) : ("Duplicate scaffold found: "+r1.id+"\n"+
"Please ensure all scaffolds have unique names (using rename.sh), or add the flag 'renameduplicates'");
System.err.println("WARNING: Duplicate scaffold is being renamed; future warnings will be suppressed."
+ "\nTo force a crash when this occurs, add the flag 'renameduplicates=f'\nDuplicate name: "+r1.id);
}
warned=true;
x=nameMap.get(name);
}
assert(x==null || useRefNames) : "Duplicate scaffold name "+r1.id+" found in scafnum "+x+" and "+scaffoldNames.size();
if(x!=null){
id=x;
}else{
id=scaffoldNames.size();
scaffoldNames.add(name);
Integer old=nameMap.put(name, id);
}
}
if(useRefNames){assert(refNum==id);}
refScafCounts[refNum]++;
int len=r1.pairLength();
r1.obj=id;
if(r2!=null){r2.obj=id;}
scaffoldLengths.increment(id, len);
}
if(REPLICATE_AMBIGUOUS){
reads2=Tools.replicateAmbiguous(reads2, k);
}
/* Send a pointer to the read list to each LoadThread */
for(LoadThread lt : loaders){
boolean b=true;
while(b){
try {
lt.queue.put(reads2);
b=false;
} catch (InterruptedException e) {
//TODO: This will hang due to still-running threads.
throw new RuntimeException(e);
}
}
}
}
/* Dispose of the old list and fetch a new one */
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
/* Cleanup */
cris.returnList(ln);
errorState|=ReadWrite.closeStream(cris);
refNum++;
}
}
// int refNum=0;
// if(ref!=null){
// for(String refname : ref){
//
// /* Start an input stream */
// FileFormat ff=FileFormat.testInput(refname, FileFormat.FASTA, null, true, true);
// ConcurrentReadInputStream cris=ConcurrentReadInputStream.getReadInputStream(-1L, false, ff, null, null, null, Shared.USE_MPI, true);
// cris.start(); //4567
// ListNum<Read> ln=cris.nextList();
// ArrayList<Read> reads=(ln!=null ? ln.list : null);
//
// final String core=ReadWrite.stripToCore(refname);
//
// /* Iterate through read lists from the input stream */
// while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
// {
// /* Assign a unique ID number to each scaffold */
// ArrayList<Read> reads2=new ArrayList<Read>(reads);
// for(Read r1 : reads2){
// final Read r2=r1.mate;
// final Integer id=scaffoldNames.size();
// refScafCounts[refNum]++;
// scaffoldNames.add(r1.id==null ? id.toString() : r1.id);
// int len=r1.length();
// r1.obj=id;
// if(r2!=null){
// r2.obj=id;
// len+=r2.length();
// }
// scaffoldLengths.add(len);
// }
//
// if(REPLICATE_AMBIGUOUS){
// reads2=Tools.replicateAmbiguous(reads2, k);
// }
//
// /* Send a pointer to the read list to each LoadThread */
// for(LoadThread lt : loaders){
// boolean b=true;
// while(b){
// try {
// lt.queue.put(reads2);
// b=false;
// } catch (InterruptedException e) {
// //TODO: This will hang due to still-running threads.
// throw new RuntimeException(e);
// }
// }
// }
// }
//
// /* Dispose of the old list and fetch a new one */
// cris.returnList(ln);
// ln=cris.nextList();
// reads=(ln!=null ? ln.list : null);
// }
// /* Cleanup */
// cris.returnList(ln);
// errorState|=ReadWrite.closeStream(cris);
// refNum++;
// }
// }
/* If there are literal sequences to use as references */
if(literal!=null){
ArrayList<Read> list=new ArrayList<Read>(literal.length);
if(verbose){outstream.println("Adding literals "+Arrays.toString(literal));}
/* Assign a unique ID number to each literal sequence */
if(useRefNames){
final Integer id=scaffoldNames.size();
scaffoldNames.add("literal");
for(int i=0; i<literal.length; i++){
final Read r=new Read(literal[i].getBytes(), null, id);
refScafCounts[refNum]++;
scaffoldLengths.increment(id, r.length());
r.obj=id;
list.add(r);
}
}else{
for(int i=0; i<literal.length; i++){
final int id=scaffoldNames.size();
final Read r=new Read(literal[i].getBytes(), null, id);
refScafCounts[refNum]++;
scaffoldNames.add(""+id);
scaffoldLengths.set(id, r.length());
r.obj=id;
list.add(r);
}
}
if(REPLICATE_AMBIGUOUS){
list=Tools.replicateAmbiguous(list, k);
}
/* Send a pointer to the read list to each LoadThread */
for(LoadThread lt : loaders){
boolean b=true;
while(b){
try {
lt.queue.put(list);
b=false;
} catch (InterruptedException e) {
//TODO: This will hang due to still-running threads.
throw new RuntimeException(e);
}
}
}
}
/* Signal loaders to terminate */
for(LoadThread lt : loaders){
boolean b=true;
while(b){
try {
lt.queue.put(POISON);
b=false;
} catch (InterruptedException e) {
//TODO: This will hang due to still-running threads.
throw new RuntimeException(e);
}
}
}
/* Wait for loaders to die, and gather statistics */
boolean success=true;
for(LoadThread lt : loaders){
while(lt.getState()!=Thread.State.TERMINATED){
try {
lt.join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
added+=lt.addedT;
refKmers+=lt.refKmersT;
refBases+=lt.refBasesT;
refReads+=lt.refReadsT;
success&=lt.success;
}
if(!success){KillSwitch.kill("Failed loading ref kmers; aborting.");}
//Correct statistics for number of threads, since each thread processes all reference data
refKmers/=WAYS;
refBases/=WAYS;
refReads/=WAYS;
scaffoldReadCounts=new AtomicLongArray(scaffoldNames.size());
scaffoldFragCounts=new AtomicLongArray(scaffoldNames.size());
scaffoldBaseCounts=new AtomicLongArray(scaffoldNames.size());
scaffoldAmbigReadCounts=new AtomicLongArray(scaffoldNames.size());
t.stop();
if(DISPLAY_PROGRESS){
outstream.println("Added "+added+" kmers; time: \t"+t);
Shared.printMemory();
outstream.println();
}
if(verbose){
TextStreamWriter tsw=new TextStreamWriter("stdout", false, false, false, FileFormat.TEXT);
tsw.start();
for(AbstractKmerTable table : keySets){
table.dumpKmersAsText(tsw, k, 1, Integer.MAX_VALUE);
}
tsw.poisonAndWait();
}
FASTQ.PARSE_CUSTOM=oldParseCustom;
return added;
}
/**
* Match reads against reference kmers, using multiple ProcessThread.
* @param t
*/
private void spawnProcessThreads(Timer t){
t.start();
/* Create read input stream */
final ConcurrentReadInputStream cris;
final boolean paired;
{
FileFormat ff1=FileFormat.testInput(in1, FileFormat.FASTQ, null, true, true);
FileFormat ff2=FileFormat.testInput(in2, FileFormat.FASTQ, null, true, true);
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, ff1.samOrBam(), ff1, ff2, qfin1, qfin2);
cris.setSampleRate(samplerate, sampleseed);
cris.start(); //4567
paired=cris.paired();
if(!ff1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
}
/* Create read output streams */
final ConcurrentReadOutputStream rosm, rosu;
final MultiCros mcros;
if(outu1!=null){
final int buff=(!ordered ? 12 : Tools.max(32, 2*Shared.threads()));
FileFormat ff1=FileFormat.testOutput(outu1, FileFormat.FASTQ, null, true, overwrite, append, ordered);
FileFormat ff2=FileFormat.testOutput(outu2, FileFormat.FASTQ, null, true, overwrite, append, ordered);
rosu=ConcurrentReadOutputStream.getStream(ff1, ff2, null, null, buff, null, true);
rosu.start();
}else{rosu=null;}
if(outm1!=null){
final int buff=(!ordered ? 12 : Tools.max(32, 2*Shared.threads()));
FileFormat ff1=FileFormat.testOutput(outm1, FileFormat.FASTQ, null, true, overwrite, append, ordered);
FileFormat ff2=FileFormat.testOutput(outm2, FileFormat.FASTQ, null, true, overwrite, append, ordered);
rosm=ConcurrentReadOutputStream.getStream(ff1, ff2, null, null, buff, null, true);
rosm.start();
}else{rosm=null;}
if(outpattern!=null){
final int buff=(!ordered ? 12 : Tools.max(32, 2*Shared.threads()));
mcros=new MultiCros(outpattern, null, ordered, overwrite, append, true, false, FileFormat.FASTQ, buff);
}else{mcros=null;}
if(rosu!=null || rosm!=null || mcros!=null){
t.stop();
outstream.println("Started output streams:\t"+t);
t.start();
}
/* Optionally skip the first reads, since initial reads may have lower quality */
if(skipreads>0){
long skipped=0;
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
while(skipped<skipreads && reads!=null && reads.size()>0){
skipped+=reads.size();
if(rosm!=null){rosm.add(new ArrayList<Read>(1), ln.id);}
if(rosu!=null){rosu.add(new ArrayList<Read>(1), ln.id);}
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
cris.returnList(ln);
if(reads==null || reads.isEmpty()){
ReadWrite.closeStreams(cris, rosu, rosm);
ReadWrite.closeStreams(mcros);
outstream.println("Skipped all of the reads.");
System.exit(0);
}
}
/* Create ProcessThreads */
ArrayList<ProcessThread> alpt=new ArrayList<ProcessThread>(THREADS);
for(int i=0; i<THREADS; i++){alpt.add(new ProcessThread(cris, rosm, rosu, mcros, ALLOW_LOCAL_ARRAYS));}
for(ProcessThread pt : alpt){pt.start();}
/* Wait for threads to die, and gather statistics */
for(ProcessThread pt : alpt){
while(pt.getState()!=Thread.State.TERMINATED){
try {
pt.join();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
readsIn+=pt.readsInT;
fragsIn+=pt.fragsInT;
basesIn+=pt.basesInT;
readsMatched+=pt.readsMatchedT;
basesMatched+=pt.basesMatchedT;
readsUnmatched+=pt.readsUnmatchedT;
basesUnmatched+=pt.basesUnmatchedT;
readsQTrimmed+=pt.readsQTrimmedT;
basesQTrimmed+=pt.basesQTrimmedT;
readsFTrimmed+=pt.readsFTrimmedT;
basesFTrimmed+=pt.basesFTrimmedT;
readsQFiltered+=pt.readsQFilteredT;
basesQFiltered+=pt.basesQFilteredT;
correctReads+=pt.correctT;
incorrectReads+=pt.incorrectT;
if(pt.scaffoldReadCountsT!=null && scaffoldReadCounts!=null){
for(int i=0; i<pt.scaffoldReadCountsT.length; i++){scaffoldReadCounts.addAndGet(i, pt.scaffoldReadCountsT[i]);}
pt.scaffoldReadCountsT=null;
}
if(pt.scaffoldBaseCountsT!=null && scaffoldBaseCounts!=null){
for(int i=0; i<pt.scaffoldBaseCountsT.length; i++){scaffoldBaseCounts.addAndGet(i, pt.scaffoldBaseCountsT[i]);}
pt.scaffoldBaseCountsT=null;
}
if(pt.scaffoldFragCountsT!=null && scaffoldFragCounts!=null){
for(int i=0; i<pt.scaffoldFragCountsT.length; i++){scaffoldFragCounts.addAndGet(i, pt.scaffoldFragCountsT[i]);}
pt.scaffoldFragCountsT=null;
}
if(pt.scaffoldAmbigReadCountsT!=null && scaffoldAmbigReadCounts!=null){
for(int i=0; i<pt.scaffoldAmbigReadCountsT.length; i++){scaffoldAmbigReadCounts.addAndGet(i, pt.scaffoldAmbigReadCountsT[i]);}
pt.scaffoldAmbigReadCountsT=null;
}
if(barcodeData!=null) {
barcodeData.merge(pt.barcodeDataT);
}
}
/* Shut down I/O streams; capture error status */
errorState|=ReadWrite.closeStreams(cris, rosu, rosm);
errorState|=ReadWrite.closeStreams(mcros);
errorState|=ReadStats.writeAll();
t.stop();
if(showSpeed){
outstream.println("Processing time: \t\t"+t);
}
}
/*--------------------------------------------------------------*/
/*---------------- Inner Classes ----------------*/
/*--------------------------------------------------------------*/
/**
* Loads kmers into a table. Each thread handles all kmers X such that X%WAYS==tnum.
*/
private class LoadThread extends Thread{
public LoadThread(final int tnum_){
tnum=tnum_;
map=keySets[tnum];
}
/**
* Get the next list of reads (or scaffolds) from the queue.
* @return List of reads
*/
private ArrayList<Read> fetch(){
ArrayList<Read> list=null;
while(list==null){
try {
list=queue.take();
} catch (InterruptedException e) {
// TODO Auto-generated catch block
e.printStackTrace();
}
}
return list;
}
@Override
public void run(){
ArrayList<Read> reads=fetch();
while(reads!=POISON){
for(Read r1 : reads){
assert(r1.pairnum()==0);
final Read r2=r1.mate;
addedT+=addToMap(r1, refSkip);
if(r2!=null){
addedT+=addToMap(r2, refSkip);
}
}
reads=fetch();
}
// if(AbstractKmerTable.TESTMODE){
// for(int i=0; i<ll.size; i++){
// assert(map.contains(ll.get(i), il.get(i)));
// assert(!map.contains(ll.get(i), Integer.MAX_VALUE));
// }
// ll=null;
// il=null;
// }
if(map.canRebalance() && map.size()>2L*map.arrayLength()){
map.rebalance();
}
success=true;
}
/**
* @param r The current read to process
* @param skip Number of bases to skip between kmers
* @return Number of kmers stored
*/
private long addToMap(final Read r, final int skip){
final byte[] bases=r.bases;
final int shift=2*k;
final int shift2=shift-2;
final long mask=(shift>63 ? -1L : ~((-1L)<<shift));
final long kmask=lengthMasks[k];
long kmer=0;
long rkmer=0;
long added=0;
int len=0;
int totalKmers=0;
if(tnum==0){
if(storeRefBases){
assert(r.mate==null);
assert(scaffolds.size()==(Integer)r.obj) : scaffolds.size()+", "+(Integer)r.obj/*+"\n"+r.toFasta()*/;
scaffolds.add(bases);
}
if(bases==null || bases.length<k){scaffoldKmers.add(0);}
}
if(bases!=null){
refReadsT++;
refBasesT+=bases.length;
}
if(bases==null || bases.length<k){return 0;}
final int id=(Integer)r.obj;
if(skip>1){ //Process while skipping some kmers
for(int i=0; i<bases.length; i++){
byte b=bases[i];
long x=AminoAcid.baseToNumber[b];
long x2=AminoAcid.baseToComplementNumber[b];
kmer=((kmer<<2)|x)&mask;
rkmer=((rkmer>>>2)|(x2<<shift2))&mask;
if(x<0){len=0; rkmer=0;}else{len++;}
if(verbose){outstream.println("Scanning1 i="+i+", kmer="+kmer+", rkmer="+rkmer+", bases="+new String(bases, Tools.max(0, i-k2), Tools.min(i+1, k)));}
if(len>=k){
totalKmers++;
if(len%skip==0){
final long extraBase=(i>=bases.length-1 ? -1 : AminoAcid.baseToNumber[bases[i+1]]);
added+=addToMap(kmer, rkmer, k, extraBase, id, kmask);
}
}
}
}else{ //Process all kmers
for(int i=0; i<bases.length; i++){
byte b=bases[i];
long x=AminoAcid.baseToNumber[b];
long x2=AminoAcid.baseToComplementNumber[b];
kmer=((kmer<<2)|x)&mask;
rkmer=((rkmer>>>2)|(x2<<shift2))&mask;
if(x<0){len=0; rkmer=0;}else{len++;}
if(verbose){outstream.println("Scanning2 i="+i+", kmer="+kmer+", rkmer="+rkmer+", bases="+new String(bases, Tools.max(0, i-k2), Tools.min(i+1, k)));}
if(len>=k){
totalKmers++;
final long extraBase=(i>=bases.length-1 ? -1 : AminoAcid.baseToNumber[bases[i+1]]);
final long atm=addToMap(kmer, rkmer, k, extraBase, id, kmask);
added+=atm;
}
}
}
refKmersT+=totalKmers;
if(tnum==0){scaffoldKmers.add(totalKmers);}
return added;
}
/**
* Adds this kmer to the table, including any mutations implied by editDistance or hammingDistance.
* @param kmer Forward kmer
* @param rkmer Reverse kmer
* @param extraBase Base added to end in case of deletions
* @param id Scaffold number
* @param kmask0
* @return Number of kmers stored
*/
private long addToMap(final long kmer, final long rkmer, final int len, final long extraBase, final int id, final long kmask0){
assert(kmask0==lengthMasks[len]) : kmask0+", "+len+", "+lengthMasks[len]+", "+Long.numberOfTrailingZeros(kmask0)+", "+Long.numberOfTrailingZeros(lengthMasks[len]);
if(verbose){outstream.println("addToMap_A; len="+len+"; kMasks[len]="+lengthMasks[len]);}
assert((kmer&kmask0)==0);
final long added;
if(hammingDistance==0){
final long key=toValue(kmer, rkmer, kmask0);
if(failsSpeed(key)){return 0;}
if(key%WAYS!=tnum){return 0;}
if(verbose){outstream.println("addToMap_B: "+AminoAcid.kmerToString(kmer&~lengthMasks[len], len)+" = "+key);}
// int[] old=map.getValues(key, new int[1]);
// int[] old=map.getValues(key, new int[1]); //123
added=map.set(key, id);
// assert(old==null || map.contains(key, old)); //123
// assert(map.contains(key, id)); //123
// ll.add(key);
// il.add(id); assert(AbstractKmerTable.TESTMODE);
// if(AbstractKmerTable.TESTMODE){
// for(int i=0; i<ll.size; i++){
// assert(map.contains(ll.get(i), il.get(i)));
// assert(!map.contains(ll.get(i), Integer.MAX_VALUE));
// }
// }
}else if(editDistance>0){
// long extraBase=(i>=bases.length-1 ? -1 : AminoAcid.baseToNumber[bases[i+1]]);
added=mutate(kmer, rkmer, len, id, editDistance, extraBase);
}else{
added=mutate(kmer, rkmer, len, id, hammingDistance, -1);
}
if(verbose){outstream.println("addToMap added "+added+" keys.");}
return added;
}
// private LongList ll=new LongList();
// private IntList il=new IntList();
/**
* Mutate and store this kmer through 'dist' recursions.
* @param kmer Forward kmer
* @param rkmer Reverse kmer
* @param id Scaffold number
* @param dist Number of mutations
* @param extraBase Base added to end in case of deletions
* @return Number of kmers stored
*/
private long mutate(final long kmer, final long rkmer, final int len, final int id, final int dist, final long extraBase){
long added=0;
final long key=toValue(kmer, rkmer, lengthMasks[len]);
if(verbose){outstream.println("mutate_A; len="+len+"; kmer="+kmer+"; rkmer="+rkmer+"; kMasks[len]="+lengthMasks[len]);}
if(key%WAYS==tnum){
if(verbose){outstream.println("mutate_B: "+AminoAcid.kmerToString(kmer&~lengthMasks[len], len)+" = "+key);}
int x=map.set(key, id);
if(verbose){outstream.println("mutate_B added "+x+" keys.");}
added+=x;
assert(map.contains(key));
}
if(dist>0){
final int dist2=dist-1;
//Sub
for(int j=0; j<4; j++){
for(int i=0; i<len; i++){
final long temp=(kmer&clearMasks[i])|setMasks[j][i];
if(temp!=kmer){
long rtemp=rcomp(temp, len);
added+=mutate(temp, rtemp, len, id, dist2, extraBase);
}
}
}
if(editDistance>0){
//Del
if(extraBase>=0 && extraBase<=3){
for(int i=1; i<len; i++){
final long temp=(kmer&leftMasks[i])|((kmer<<2)&rightMasks[i])|extraBase;
if(temp!=kmer){
long rtemp=rcomp(temp, len);
added+=mutate(temp, rtemp, len, id, dist2, -1);
}
}
}
//Ins
final long eb2=kmer&3;
for(int i=1; i<len; i++){
final long temp0=(kmer&leftMasks[i])|((kmer&rightMasks[i])>>2);
for(int j=0; j<4; j++){
final long temp=temp0|setMasks[j][i-1];
if(temp!=kmer){
long rtemp=rcomp(temp, len);
added+=mutate(temp, rtemp, len, id, dist2, eb2);
}
}
}
}
}
return added;
}
/*--------------------------------------------------------------*/
/** Number of kmers stored by this thread */
public long addedT=0;
/** Number of items encountered by this thread */
public long refKmersT=0, refReadsT=0, refBasesT=0;
/** Thread number; used to determine which kmers to store */
public final int tnum;
/** Buffer of input read lists */
public final ArrayBlockingQueue<ArrayList<Read>> queue=new ArrayBlockingQueue<ArrayList<Read>>(32);
/** Destination for storing kmers */
private final AbstractKmerTable map;
/** Completed successfully */
boolean success=false;
}
/*--------------------------------------------------------------*/
/**
* Matches read kmers against reference kmers, performs binning and/or trimming, and writes output.
*/
private class ProcessThread extends Thread{
/**
* Constructor
* @param cris_ Read input stream
* @param rosu_ Unmatched read output stream (optional)
* @param rosm_ Matched read output stream (optional)
*/
public ProcessThread(ConcurrentReadInputStream cris_, ConcurrentReadOutputStream rosm_, ConcurrentReadOutputStream rosu_,
MultiCros mcros_, boolean localArrays){
cris=cris_;
rosm=rosm_;
rosu=rosu_;
mcros=mcros_;
readstats=(ReadStats.collectingStats()) ? new ReadStats() : null;
barcodeDataT=(trackBarcodes ? new BarcodeMappingStats() : null);
final int alen=(scaffoldNames==null ? 0 : scaffoldNames.size());
if(localArrays && alen>0 && alen<10000){
scaffoldReadCountsT=new long[alen];
scaffoldBaseCountsT=new long[alen];
scaffoldFragCountsT=new long[alen];
scaffoldAmbigReadCountsT=new long[alen];
}else{
scaffoldReadCountsT=scaffoldBaseCountsT=scaffoldFragCountsT=scaffoldAmbigReadCountsT=null;
}
if(USE_COUNTVECTOR){
countVector=new IntList(1000);
countArray=null;
}else{
countVector=null;
countArray=new int[alen];
}
}
@Override
public void run(){
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
final ArrayList<Read> mlist=(rosm==null ? null : new ArrayList<Read>(Shared.bufferLen()));
final ArrayList<Read> ulist=(rosu==null ? null : new ArrayList<Read>(Shared.bufferLen()));
final ArrayListSet als=(outpattern==null ? null : new ArrayListSet(ordered));
//While there are more reads lists...
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
//For each read (or pair) in the list...
for(int i=0; i<reads.size(); i++){
final Read r1=reads.get(i);
final Read r2=r1.mate;
if(!r1.validated()){r1.validate(true);}
if(r2!=null && !r2.validated()){r2.validate(true);}
if(readstats!=null){
readstats.addToHistograms(r1);
}
final int initialLength1=r1.length();
final int initialLength2=(r1.mateLength());
final int minlen1=(int)Tools.max(initialLength1*minLenFraction, minReadLength);
final int minlen2=(int)Tools.max(initialLength2*minLenFraction, minReadLength);
if(loglog!=null){loglog.hash(r1);}
if(verbose){outstream.println("Considering read "+r1.id+" "+new String(r1.bases));}
fragsInT++;
readsInT+=(1+r1.mateCount());
basesInT+=r1.pairLength();
boolean remove=false;
if(chastityFilter){
if(r1!=null && r1.failsChastity()){
basesQFilteredT+=r1.length();
readsQFilteredT++;
r1.setDiscarded(true);
}
if(r2!=null && r2.failsChastity()){
basesQFilteredT+=r2.length();
readsQFilteredT++;
r2.setDiscarded(true);
}
}
if(forceTrimLeft>0 || forceTrimRight>0 || forceTrimRight2>0 || forceTrimModulo>0){
if(r1!=null && !r1.discarded()){
final int len=r1.length();
final int a=forceTrimLeft>0 ? forceTrimLeft : 0;
final int b0=forceTrimModulo>0 ? len-1-len%forceTrimModulo : len;
final int b1=forceTrimRight>0 ? forceTrimRight : len;
final int b2=forceTrimRight2>0 ? len-1-forceTrimRight2 : len;
final int b=Tools.min(b0, b1, b2);
final int x=TrimRead.trimToPosition(r1, a, b, 1);
basesFTrimmedT+=x;
readsFTrimmedT+=(x>0 ? 1 : 0);
if(r1.length()<minlen1){r1.setDiscarded(true);}
}
if(r2!=null && !r2.discarded()){
final int len=r2.length();
final int a=forceTrimLeft>0 ? forceTrimLeft : 0;
final int b0=forceTrimModulo>0 ? len-1-len%forceTrimModulo : len;
final int b1=forceTrimRight>0 ? forceTrimRight : len;
final int b2=forceTrimRight2>0 ? len-1-forceTrimRight2 : len;
final int b=Tools.min(b0, b1, b2);
final int x=TrimRead.trimToPosition(r2, a, b, 1);
basesFTrimmedT+=x;
readsFTrimmedT+=(x>0 ? 1 : 0);
if(r2.length()<minlen2){r2.setDiscarded(true);}
}
}
if(removePairsIfEitherBad){remove=r1.discarded() || (r2!=null && r2.discarded());}
else{remove=r1.discarded() && (r2==null || r2.discarded());}
if(!remove){
//Do quality trimming
int rlen1=0, rlen2=0;
if(r1!=null){
if(qtrimLeft || qtrimRight){
int x=TrimRead.trimFast(r1, qtrimLeft, qtrimRight, trimq, trimE, 1);
basesQTrimmedT+=x;
readsQTrimmedT+=(x>0 ? 1 : 0);
}
rlen1=r1.length();
if(rlen1<minlen1 || rlen1>maxReadLength){
r1.setDiscarded(true);
if(verbose){outstream.println(r1.id+" discarded due to length.");}
}
}
if(r2!=null){
if(qtrimLeft || qtrimRight){
int x=TrimRead.trimFast(r2, qtrimLeft, qtrimRight, trimq, trimE, 1);
basesQTrimmedT+=x;
readsQTrimmedT+=(x>0 ? 1 : 0);
}
rlen2=r2.length();
if(rlen2<minlen2 || rlen2>maxReadLength){
r2.setDiscarded(true);
if(verbose){outstream.println(r2.id+" discarded due to length.");}
}
}
//Discard reads if too short
if((removePairsIfEitherBad && (r1.discarded() || (r2!=null && r2.discarded()))) ||
(r1.discarded() && (r2==null || r2.discarded()))){
basesQFilteredT+=r1.pairLength();
readsQTrimmedT+=r1.pairCount();
remove=true;
}
}
if(!remove){
//Do quality filtering
//Determine whether to discard the reads based on average quality
if(minAvgQuality>0){
if(r1!=null && r1.quality!=null && r1.avgQuality(false, minAvgQualityBases)<minAvgQuality){
r1.setDiscarded(true);
if(verbose){outstream.println(r1.id+" discarded due to low quality.");}
}
if(r2!=null && r2.quality!=null && r2.avgQuality(false, minAvgQualityBases)<minAvgQuality){
r2.setDiscarded(true);
if(verbose){outstream.println(r2.id+" discarded due to low quality.");}
}
}
//Determine whether to discard the reads based on the presence of Ns
if(maxNs>=0){
if(r1!=null && r1.countUndefined()>maxNs){
r1.setDiscarded(true);
if(verbose){outstream.println(r1.id+" discarded due to Ns.");}
}
if(r2!=null && r2.countUndefined()>maxNs){
r2.setDiscarded(true);
if(verbose){outstream.println(r2.id+" discarded due to Ns.");}
}
}
//Determine whether to discard the reads based on a lack of useful kmers
if(minConsecutiveBases>0){
if(r1!=null && !r1.discarded() && !r1.hasMinConsecutiveBases(minConsecutiveBases)){r1.setDiscarded(true);}
if(r2!=null && !r2.discarded() && !r2.hasMinConsecutiveBases(minConsecutiveBases)){r2.setDiscarded(true);}
}
//Discard reads if too short
if((removePairsIfEitherBad && (r1.discarded() || (r2!=null && r2.discarded()))) ||
(r1.discarded() && (r2==null || r2.discarded()))){
basesQFilteredT+=r1.pairLength();
readsQFilteredT+=r1.pairCount();
remove=true;
}
}
final int sites, assigned;
if(remove){
if(r1!=null){
basesQFilteredT+=r1.length();
readsQFilteredT++;
}
if(r2!=null){
basesQFilteredT+=r2.length();
readsQFilteredT++;
}
sites=assigned=0;
}else{
if(ecc && r1!=null && r2!=null){BBMerge.findOverlapStrict(r1, r2, true);}
//Do kmer matching
if(keepPairsTogether){
final int a, b, max;
if(countArray==null){
countVector.size=0;
a=findBestMatch(r1, keySets, countVector);
b=findBestMatch(r2, keySets, countVector);
if(verbose){outstream.println("countVector: "+countVector);}
max=condenseLoose(countVector, idList1, countList1);
}else{
idList1.size=0;
a=findBestMatch(r1, keySets, countArray, idList1);
b=findBestMatch(r2, keySets, countArray, idList1);
max=condenseLoose(countArray, idList1, countList1);
}
if(verbose){
outstream.println("idList1: "+idList1);
outstream.println("countList1: "+countList1);
}
final int cz=clearzoneFraction<=0 ? clearzone :
Tools.max(clearzone, (int)Math.ceil(clearzoneFraction*(r1.numValidPairKmers(k))));
filterTopScaffolds(r1, r2, idList1, countList1, finalList1, max, cz);
if(verbose){
outstream.println("idList1: "+idList1);
outstream.println("countList1: "+countList1);
outstream.println("finalList1: "+finalList1);
}
sites=finalList1.size;
final int minhits=Tools.max(minKmerHits, (int)(minKmerFraction*numKmers(r1, r2, k)));
if(max>=minhits){
assigned=assignTogether(r1, r2, als);
}else{
readsUnmatchedT+=r1.pairCount();
basesUnmatchedT+=r1.pairLength();
assigned=0;
}
if(rename){
rename(r1, idList1, countList1, max, assigned);
rename(r2, idList1, countList1, max, assigned);
}
}else{
final int max1, max2, a, b;
{
if(countArray==null){
countVector.size=0;
a=findBestMatch(r1, keySets, countVector);
max1=condenseLoose(countVector, idList1, countList1);
}else{
idList1.size=0;
a=findBestMatch(r1, keySets, countArray, idList1);
max1=condenseLoose(countArray, idList1, countList1);
}
// if(rename){rename(r1, idList1, countList1, max1);}
final int cz=clearzoneFraction<=0 ? clearzone :
Tools.max(clearzone, (int)Math.ceil(clearzoneFraction*(r1.numValidKmers(k))));
filterTopScaffolds(r1, null, idList1, countList1, finalList1, max1, cz);
}
if(r2!=null){
if(countArray==null){
countVector.size=0;
b=findBestMatch(r2, keySets, countVector);
max2=condenseLoose(countVector, idList2, countList1);
}else{
idList2.size=0;
b=findBestMatch(r2, keySets, countArray, idList2);
max2=condenseLoose(countArray, idList2, countList2);
}
// if(rename){rename(r2, idList2, countList2, max2);}
final int cz=clearzoneFraction<=0 ? clearzone :
Tools.max(clearzone, (int)Math.ceil(clearzoneFraction*(r2.numValidKmers(k))));
filterTopScaffolds(r2, null, idList2, countList2, finalList2, max2, cz);
}else{max2=0;}
sites=finalList1.size+finalList2.size;
assigned=assignIndependently(r1, r2, max1, max2, als);
if(rename) {
rename(r1, idList1, countList1, max1, assigned);
if(r2!=null) {rename(r2, idList2, countList2, max2, assigned);}
}
}
}
if(remove || assigned<1){
if(ulist!=null){
if(loglogOut!=null){loglogOut.hash(r1);}
ulist.add(r1);
}
}else{
if(mlist!=null){mlist.add(r1);}
}
}
//Send matched list to matched output stream
if(rosu!=null){
rosu.add(ulist, ln.id);
ulist.clear();
}
//Send unmatched list to unmatched output stream
if(rosm!=null){
rosm.add(mlist, ln.id);
mlist.clear();
}
if(mcros!=null){
mcros.add(als, ln.id);
}
//Fetch a new read list
cris.returnList(ln);
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
cris.returnList(ln);
}
/*--------------------------------------------------------------*/
/*---------------- Helper Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* @param r
* @param idList
* @param countList
*/
private void rename(Read r, IntList idList, IntList countList, final int max, int assignedSites) {
if(r==null || (idList.size<1 && !addUnknown)){return;}
StringBuilder sb=new StringBuilder();
if(r.id==null){sb.append(r.numericID);}
else{sb.append(r.id);}
if(idList.size>0 && (assignedSites>0 || !topHitOnly)) {
for(int i=0; i<idList.size; i++){
int id=idList.get(i);
int count=countList.get(i);
if(count==max || !topHitOnly) {
sb.append('\t');
sb.append(scaffoldNames.get(id));
if(addCount) {
sb.append('=');
sb.append(count);
}
if(topHitOnly) {break;}
}
}
}else if(addUnknown) {
sb.append('\t').append("unknown");
}
r.id=sb.toString();
}
private void addGeneSet(IntList finalList1, int start, int stop, int readSum, int lenSum) {
if(MAKE_GENE_SETS && stop>start+1) {
finalList1.sort();
ByteBuilder bb=new ByteBuilder();
int len=0;
for(int j=start; j<stop; j++) {
int id=finalList1.get(j);
if(!bb.isEmpty()) {bb.comma();}
bb.append(scaffoldNames.get(id));
len+=scaffoldLengths.get(id);
}
String name=bb.toString();
StringCount sc=setMap.get(name);
if(sc==null) {
synchronized(setMap) {
sc=setMap.get(name);
if(sc==null) {
sc=new StringCount(name);
sc.length=len;
setMap.put(name, sc);
}
}
}
assert(sc!=null);
synchronized(sc) {
sc.reads+=readSum;
sc.bases+=lenSum;
sc.ambigReads+=readSum;
}
}
}
/**
* @param r1 Read 1
* @param r2 Read 2
* @return Number of sites assigned
*/
private int assignTogether(Read r1, Read r2, ArrayListSet als){
final int sites=finalList1.size;
final int lenSum=r1.length()+(r1.mateLength());
final int readSum=1+(r2==null ? 0 : 1);
final int start, stop;
final String barcode=(trackBarcodes ? r1.barcode(true) : null);
if(sites<2 || ambigMode==AMBIG_ALL){
start=0;
stop=sites;
}else if(ambigMode==AMBIG_TOSS){
start=stop=0;
}else if(ambigMode==AMBIG_FIRST){
finalList1.sort();
start=0;
stop=1;
}else if(ambigMode==AMBIG_RANDOM){
start=(int)(r1.numericID%sites);
stop=start+1;
}else{
throw new RuntimeException("Unknown mode "+ambigMode);
}
if(MAKE_GENE_SETS && stop>start+1) {addGeneSet(finalList1, start, stop, readSum, lenSum);}
for(int j=start; j<stop; j++){
int id=finalList1.get(j);
if(als!=null){
als.add(r1, scaffoldNames.get(id));
}
if(parsecustom && j==start){
String scafName=scaffoldNames.get(id);
String rname=r1.parseCustomRname();
if(scafName.equals(rname)){
correctT+=(1+r1.mateCount());
}else{
incorrectT+=(1+r1.mateCount());
}
}
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, scaffoldNames.get(id), r1.pairCount());}
if(scaffoldReadCountsT!=null){
scaffoldReadCountsT[id]+=readSum;
scaffoldBaseCountsT[id]+=lenSum;
scaffoldFragCountsT[id]++;
if(sites>1) {scaffoldAmbigReadCountsT[id]+=readSum;}
}else{
scaffoldReadCounts.addAndGet(id, readSum);
scaffoldBaseCounts.addAndGet(id, lenSum);
scaffoldFragCounts.addAndGet(id, 1);
if(sites>1) {scaffoldAmbigReadCounts.addAndGet(id, readSum);}
}
}
if(trackBarcodes){barcodeDataT.incrementCodeMap(barcode, r1.pairCount());}
if(start<stop){
readsMatchedT+=r1.pairCount();
basesMatchedT+=r1.pairLength();
}else{
readsUnmatchedT+=r1.pairCount();
basesUnmatchedT+=r1.pairLength();
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, sites>1 ? "AMBIG" : "UNKNOWN", r1.pairCount());}
}
return stop-start;
}
/**
* @param r1 Read 1
* @param r2 Read 2
* @param max1 Highest match count for read 1
* @param max2 Highest match count for read 2
* @return Number of sites assigned
*/
private int assignIndependently(Read r1, Read r2, int max1, int max2, ArrayListSet als){
assert(als==null || r2==null) : "Pattern output does not work with keepPairsTogether=false and paired reads\n"+als;
int assigned=0;
final String barcode=(trackBarcodes ? r1.barcode(true) : null);
if(trackBarcodes){barcodeDataT.incrementCodeMap(barcode, r1.pairCount());}
if(max1>=Tools.max(minKmerHits, (int)(minKmerFraction*numKmers(r1, null, k)))){
final int sites=finalList1.size;
final int lenSum=r1.length();
final int start, stop;
if(sites<2 || ambigMode==AMBIG_ALL){
start=0;
stop=sites;
}else if(ambigMode==AMBIG_TOSS){
start=stop=0;
}else if(ambigMode==AMBIG_FIRST){
finalList1.sort();
start=0;
stop=1;
}else if(ambigMode==AMBIG_RANDOM){
start=(int)(r1.numericID%sites);
stop=start+1;
}else{
throw new RuntimeException("Unknown mode "+ambigMode);
}
if(MAKE_GENE_SETS && stop>start+1) {addGeneSet(finalList1, start, stop, 1, lenSum);}
for(int j=start; j<stop; j++){
int id=finalList1.get(j);
if(als!=null){
als.add(r1, scaffoldNames.get(id));
}
if(parsecustom && j==start){
String scafName=scaffoldNames.get(id);
String rname=r1.parseCustomRname();
if(scafName.equals(rname)){
correctT++;
}else{
incorrectT++;
}
}
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, scaffoldNames.get(id), 1);}
if(scaffoldReadCountsT!=null){
scaffoldReadCountsT[id]++;
scaffoldBaseCountsT[id]+=lenSum;
if(max1>=max2){
scaffoldFragCountsT[id]++;
}
if(sites>1) {scaffoldAmbigReadCountsT[id]++;}
}else{
scaffoldReadCounts.addAndGet(id, 1);
scaffoldBaseCounts.addAndGet(id, lenSum);
if(max1>=max2){
scaffoldFragCounts.addAndGet(id, 1);
}
if(sites>1) {scaffoldAmbigReadCounts.incrementAndGet(id);}
}
}
if(start<stop){
readsMatchedT++;
basesMatchedT+=r1.length();
assigned+=(stop-start);
}else{
readsUnmatchedT++;
basesUnmatchedT+=r1.length();
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, sites>1 ? "AMBIG" : "UNKNOWN", 1);}
}
}
if(max2>=Tools.max(minKmerHits, (int)(minKmerFraction*numKmers(r2, null, k)))){
final int sites=finalList2.size;
final int lenSum=r2.length();
final int start, stop;
if(sites<2 || ambigMode==AMBIG_ALL){
start=0;
stop=sites;
}else if(ambigMode==AMBIG_TOSS){
start=stop=0;
}else if(ambigMode==AMBIG_FIRST){
finalList2.sort();
start=0;
stop=1;
}else if(ambigMode==AMBIG_RANDOM){
start=(int)(r2.numericID%sites);
stop=start+1;
}else{
throw new RuntimeException("Unknown mode "+ambigMode);
}
if(MAKE_GENE_SETS && stop>start+1) {addGeneSet(finalList1, start, stop, 1, lenSum);}
for(int j=start; j<stop; j++){
int id=finalList2.get(j);
if(als!=null){
als.add(r2, scaffoldNames.get(id));
throw new RuntimeException("Pattern output does not currently work with keepPairsTogether=false");
}
if(parsecustom && j==start){
String scafName=scaffoldNames.get(id);
String rname=r2.parseCustomRname();
if(scafName.equals(rname)){
correctT++;
}else{
incorrectT++;
}
}
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, scaffoldNames.get(id), 1);}
if(scaffoldReadCountsT!=null){
scaffoldReadCountsT[id]++;
scaffoldBaseCountsT[id]+=lenSum;
if(max2>max1){
scaffoldFragCountsT[id]++;
}
if(sites>1) {scaffoldAmbigReadCountsT[id]++;}
}else{
scaffoldReadCounts.addAndGet(id, 1);
scaffoldBaseCounts.addAndGet(id, lenSum);
if(max2>max1){
scaffoldFragCounts.addAndGet(id, 1);
}
if(sites>1) {scaffoldAmbigReadCounts.incrementAndGet(id);}
}
}
if(start<stop){
readsMatchedT++;
basesMatchedT+=r2.length();
assigned+=(stop-start);
}else{
readsUnmatchedT++;
basesUnmatchedT+=r2.length();
if(trackBarcodes){barcodeDataT.incrementSourceMap(barcode, sites>1 ? "AMBIG" : "UNKNOWN", 1);}
}
}
return assigned;
}
/**
* Pack a list of nonunique values into a list of unique values and a list of their counts.
* @param loose Nonunique values
* @param packed Unique values
* @param counts Counts of values
* @return
*/
private int condenseLoose(IntList loose, IntList packed, IntList counts){
packed.size=0;
counts.size=0;
if(loose.size<1){return 0;}
loose.sort();
int prev=-1;
int max=0;
int count=0;
for(int i=0; i<loose.size; i++){
int id=loose.get(i);
// outstream.println("i="+i+", id="+id+", count="+count+", prev="+prev);
if(id==prev){
count++;
}else{
if(count>0){
packed.add(prev);
counts.add(count);
max=Tools.max(count, max);
// assert(false) : "i="+i+", "+id+", "+count+", "+packed+", "+counts;
}
prev=id;
count=1;
}
}
if(count>0){
packed.add(prev);
counts.add(count);
max=Tools.max(count, max);
}
return max;
}
/**
* Pack a list of counts from an array to an IntList.
* @param loose Counter array
* @param packed Unique values
* @param counts Counts of values
* @return
*/
private int condenseLoose(int[] loose, IntList packed, IntList counts){
counts.size=0;
if(packed.size<1){return 0;}
int max=0;
for(int i=0; i<packed.size; i++){
final int p=packed.get(i);
final int c=loose[p];
counts.add(c);
loose[p]=0;
max=Tools.max(max, c);
}
return max;
}
private void filterTopScaffolds(Read r1, Read r2, IntList packed, IntList counts, IntList out, int max, int cz){
out.size=0;
if(packed.size<1){return;}
if(processContainedRef){
filterTopScaffolds_withContainedRef(r1, r2, packed, counts, out);
}else{
filterTopScaffolds_withClearzone(packed, counts, out, max, cz);
}
}
private void filterTopScaffolds_withContainedRef(Read r1, Read r2, IntList packed, IntList counts, IntList out){
for(int i=0; i<packed.size; i++){
final int p=packed.get(i);
final int c=counts.get(i);
if(storeRefBases){
if(Tools.containsForward(r1.bases, scaffolds.get(p), hammingDistance)>=0 ||
(r2!=null && Tools.containsForward(r2.bases, scaffolds.get(p), hammingDistance)>=0)){
out.add(p);
}else if(rcomp && (Tools.containsReverse(r1.bases, scaffolds.get(p), hammingDistance)>=0 ||
(r2!=null && Tools.containsReverse(r2.bases, scaffolds.get(p), hammingDistance)>=0))){
out.add(p);
}
}else if(c>=scaffoldKmers.get(p)){
out.add(p);
}
}
}
private void filterTopScaffolds_withClearzone(IntList packed, IntList counts, IntList out, int max, int cz){
final int thresh=Tools.max(1, max-cz);
for(int i=0; i<packed.size; i++){
final int c=counts.get(i);
assert((c>0) && c<=max) : c+"\n"+packed+"\n"+counts;
if(c>=thresh){
out.add(packed.get(i));
}
}
}
/**
* Transforms a kmer into all canonical values for a given Hamming distance.
* Returns the related id stored in the tables.
* @param kmer Forward kmer
* @param rkmer Reverse kmer
* @param lengthMask Bitmask with single '1' set to left of kmer
* @param qPos Position of kmer in query
* @param len kmer length
* @param qHDist Hamming distance
* @param sets Kmer hash tables
* @return Value stored in table, or -1
*/
private final int[] getValues(final long kmer, final long rkmer, final long lengthMask, final int qPos, final int len, final int qHDist, final AbstractKmerTable[] sets){
if(qHDist>0){return getValuesQHD(kmer, rkmer, lengthMask, qPos, len, qHDist, sets, qhList);}
int[] ids=getValuesInner(kmer, rkmer, lengthMask, qPos, sets);
if((ids==null || ids[0]<0) && qHDist>0){
final int qHDist2=qHDist-1;
//Sub
for(int j=0; j<4; j++){
for(int i=0; i<len; i++){
final long temp=(kmer&clearMasks[i])|setMasks[j][i];
if(temp!=kmer){
long rtemp=rcomp(temp, len);
ids=getValues(temp, rtemp, lengthMask, qPos, len, qHDist2, sets);
if(ids!=null && ids[0]>-1){return ids;}
}
}
}
}
return ids;
}
private final int[] getValuesQHD(final long kmer, final long rkmer, final long lengthMask, final int qPos, final int len, final int qHDist, final AbstractKmerTable[] sets, IntList list){
assert(qHDist>0);
list.clear();
getValuesQHD_inner(kmer, rkmer, lengthMask, qPos, len, qHDist, sets, list);
if(list.size>qhdistSizeLimit){
list.sort();
list.shrinkToUnique();
}
if(list.size>0){
list.add(-1);//indicates end
return list.array;
}
return null;
}
private final void getValuesQHD_inner(final long kmer, final long rkmer, final long lengthMask, final int qPos, final int len, final int qHDist, final AbstractKmerTable[] sets, IntList list){
final int sizeLimit=10;
int[] ids=getValuesInner(kmer, rkmer, lengthMask, qPos, sets);
if(ids!=null){
for(int x : ids){
if(x<0){break;}
if(list.size>sizeLimit || !list.contains(x)){list.add(x);}
}
}
if(qHDist>0){
final int qHDist2=qHDist-1;
//Sub
for(int j=0; j<4; j++){
for(int i=0; i<len; i++){
final long temp=(kmer&clearMasks[i])|setMasks[j][i];
if(temp!=kmer){
long rtemp=rcomp(temp, len);
getValuesQHD_inner(temp, rtemp, lengthMask, qPos, len, qHDist2, sets, list);
}
}
}
}
}
/**
* Transforms a kmer into a canonical value stored in the table and search.
* @param kmer Forward kmer
* @param rkmer Reverse kmer
* @param lengthMask Bitmask with single '1' set to left of kmer
* @param qPos Position of kmer in query
* @param sets Kmer hash tables
* @return Value stored in table
*/
private final int[] getValuesInner(final long kmer, final long rkmer, final long lengthMask, final int qPos, final AbstractKmerTable[] sets){
assert(lengthMask==0 || (kmer<lengthMask && rkmer<lengthMask)) : lengthMask+", "+kmer+", "+rkmer;
if(qSkip>1 && (qPos%qSkip!=0)){return null;}
final long max=(rcomp ? Tools.max(kmer, rkmer) : kmer);
final long key=(max&middleMask)|lengthMask;
if(passesSpeed(key)){
if(verbose){outstream.println("Testing key "+key);}
AbstractKmerTable set=sets[(int)(key%WAYS)];
if(verbose){outstream.println("Found set "+set.arrayLength()+", "+set.size());}
final int[] ids=set.getValues(key, singleton);
if(verbose){outstream.println("Found array "+(ids==null ? "null" : Arrays.toString(ids)));}
return ids;
}
return null;
}
/**
* Returns number of matching kmers.
* @param r Read to process
* @param sets Kmer tables
* @return number of total kmer matches
*/
private final int findBestMatch(final Read r, final AbstractKmerTable sets[], IntList hits){
if(r==null || r.bases==null || storedKmers<1){return 0;}
final byte[] bases=r.bases;
final int minlen=k-1;
final int minlen2=(maskMiddle ? (k-midMaskLen)/2 : k);
final int shift=2*k;
final int shift2=shift-2;
final long mask=(shift>63 ? -1L : ~((-1L)<<shift));
final long kmask=lengthMasks[k];
long kmer=0;
long rkmer=0;
int found=0;
int len=0;
if(bases==null || bases.length<k){return -1;}
final int start=(restrictRight<1 ? 0 : Tools.max(0, bases.length-restrictRight));
final int stop=(restrictLeft<1 ? bases.length : Tools.min(bases.length, restrictLeft));
/* Loop through the bases, maintaining a forward and reverse kmer via bitshifts */
for(int i=start; i<stop; i++){
byte b=bases[i];
long x=AminoAcid.baseToNumber0[b];
long x2=AminoAcid.baseToComplementNumber0[b];
kmer=((kmer<<2)|x)&mask;
rkmer=((rkmer>>>2)|(x2<<shift2))&mask;
if(b=='N' && forbidNs){len=0; rkmer=0;}else{len++;}
if(verbose){outstream.println("Scanning6 i="+i+", kmer="+kmer+", rkmer="+rkmer+", bases="+new String(bases, Tools.max(0, i-k2), Tools.min(i+1, k)));}
if(len>=minlen2 && i>=minlen){
final int[] ids=getValues(kmer, rkmer, kmask, i, k, qHammingDistance, sets);
if(ids!=null && ids[0]>-1){
for(int id : ids){
if(id==-1){break;}
hits.add(id);
}
if(verbose){outstream.println("Found = "+(found+1)+"/"+minKmerHits);}
found++;
// assert(false) : (matchMode==MATCH_FIRST)+", "+(matchMode==MATCH_UNIQUE)+", "+ (ids.length==1 || ids[1]==-1);
if(matchMode==MATCH_FIRST || (matchMode==MATCH_UNIQUE && (ids.length==1 || ids[1]==-1))){break;}
}
}
}
return found;
}
/**
* Returns number of matching kmers.
* @param r Read to process
* @param sets Kmer tables
* @return number of total kmer matches
*/
private final int findBestMatch(final Read r, final AbstractKmerTable sets[], int[] hits, IntList idList){
if(r==null || r.bases==null || storedKmers<1){return 0;}
final byte[] bases=r.bases;
final int minlen=k-1;
final int minlen2=(maskMiddle ? (k-midMaskLen)/2 : k);
final int shift=2*k;
final int shift2=shift-2;
final long mask=(shift>63 ? -1L : ~((-1L)<<shift));
final long kmask=lengthMasks[k];
long kmer=0;
long rkmer=0;
int found=0;
int len=0;
if(bases==null || bases.length<k){return -1;}
final int start=(restrictRight<1 ? 0 : Tools.max(0, bases.length-restrictRight));
final int stop=(restrictLeft<1 ? bases.length : Tools.min(bases.length, restrictLeft));
/* Loop through the bases, maintaining a forward and reverse kmer via bitshifts */
for(int i=start; i<stop; i++){
byte b=bases[i];
long x=AminoAcid.baseToNumber0[b];
long x2=AminoAcid.baseToComplementNumber0[b];
kmer=((kmer<<2)|x)&mask;
rkmer=((rkmer>>>2)|(x2<<shift2))&mask;
if(b=='N' && forbidNs){len=0; rkmer=0;}else{len++;}
if(verbose){outstream.println("Scanning6 i="+i+", kmer="+kmer+", rkmer="+rkmer+", bases="+new String(bases, Tools.max(0, i-k2), Tools.min(i+1, k)));}
if(len>=minlen2 && i>=minlen){
final int[] ids=getValues(kmer, rkmer, kmask, i, k, qHammingDistance, sets);
if(ids!=null && ids[0]>-1){
for(int id : ids){
if(id==-1){break;}
hits[id]++;
if(hits[id]==1){idList.add(id);}
}
if(verbose){outstream.println("Found = "+(found+1)+"/"+minKmerHits);}
found++;
// assert(false) : (matchMode==MATCH_FIRST)+", "+(matchMode==MATCH_UNIQUE)+", "+ (ids.length==1 || ids[1]==-1);
if(matchMode==MATCH_FIRST || (matchMode==MATCH_UNIQUE && (ids.length==1 || ids[1]==-1))){break;}
}
}
}
return found;
}
/*--------------------------------------------------------------*/
/** Input read stream */
private final ConcurrentReadInputStream cris;
/** Output read streams */
private final ConcurrentReadOutputStream rosm, rosu;
/** Output pattern read streams */
private final MultiCros mcros;
private final ReadStats readstats;
private final IntList countVector;
private final int[] countArray;
private final BarcodeMappingStats barcodeDataT;
private final IntList idList1=new IntList(), idList2=new IntList();
private final IntList countList1=new IntList(), countList2=new IntList();
private final IntList finalList1=new IntList(), finalList2=new IntList();
private final IntList qhList=new IntList();
long[] scaffoldReadCountsT;
long[] scaffoldBaseCountsT;
long[] scaffoldFragCountsT;
long[] scaffoldAmbigReadCountsT;
final int[] singleton=new int[1];
private long readsInT=0;
private long fragsInT=0;
private long basesInT=0;
private long readsMatchedT=0;
private long basesMatchedT=0;
private long readsUnmatchedT=0;
private long basesUnmatchedT=0;
private long readsQTrimmedT=0;
private long basesQTrimmedT=0;
private long readsFTrimmedT=0;
private long basesFTrimmedT=0;
private long readsQFilteredT=0;
private long basesQFilteredT=0;
private long correctT=0;
private long incorrectT=0;
}
/*--------------------------------------------------------------*/
/*---------------- Static Methods ----------------*/
/*--------------------------------------------------------------*/
/** Current available memory */
private static final long freeMemory(){
Runtime rt=Runtime.getRuntime();
return rt.freeMemory();
}
/**
* Transforms a kmer into a canonical value stored in the table. Expected to be inlined.
* @param kmer Forward kmer
* @param rkmer Reverse kmer
* @param lengthMask Bitmask with single '1' set to left of kmer
* @return Canonical value
*/
private final long toValue(long kmer, long rkmer, long lengthMask){
assert(lengthMask==0 || (kmer<lengthMask && rkmer<lengthMask)) : lengthMask+", "+kmer+", "+rkmer;
long value=(rcomp ? Tools.max(kmer, rkmer) : kmer);
return (value&middleMask)|lengthMask;
}
final long rcomp(long kmer, int len){
return /*amino ? kmer :*/ AminoAcid.reverseComplementBinaryFast(kmer, len);
}
final boolean passesSpeed(long key){
return speed<1 || ((key&Long.MAX_VALUE)%17)>=speed;
}
final boolean failsSpeed(long key){
return speed>0 && ((key&Long.MAX_VALUE)%17)<speed;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** For calculating kmer cardinality */
final CardinalityTracker loglog;
final CardinalityTracker loglogOut;
/** Has this class encountered errors while processing? */
public boolean errorState=false;
/** Fraction of available memory preallocated to arrays */
private double preallocFraction=0.5;
/** Initial size of data structures */
private int initialSize=-1;
/** Default initial size of data structures */
private static final int initialSizeDefault=128000; //123
// asdf
//This needs to be redone to better track ref (file) num and scaf num.
/** Hold kmers. A kmer X such that X%WAYS=Y will be stored in keySets[Y] */
private final AbstractKmerTable[] keySets;
/** A scaffold's name is stored at scaffoldNames.get(id).
* scaffoldNames[0] is reserved, so the first id is 1. */
private final ArrayList<String> scaffoldNames=new ArrayList<String>();
/** Names of reference files (refNames[0] is valid). */
private final ArrayList<String> refNames=new ArrayList<String>();
/** Number of scaffolds per reference. */
private final int[] refScafCounts;
/** scaffoldCounts[id] stores the number of reads with kmer matches to that scaffold */
private AtomicLongArray scaffoldReadCounts;
/** scaffoldFragCounts[id] stores the number of fragments (reads or pairs) with kmer matches to that scaffold */
private AtomicLongArray scaffoldFragCounts;
/** scaffoldBaseCounts[id] stores the number of bases with kmer matches to that scaffold */
private AtomicLongArray scaffoldBaseCounts;
/** scaffoldAmbigReadCounts[id] stores the number of reads ambiguously matching that scaffold */
private AtomicLongArray scaffoldAmbigReadCounts;
/** Set to false to force threads to share atomic counter arrays. */
private boolean ALLOW_LOCAL_ARRAYS=true;
/** scaffoldLengths[id] stores the length of that scaffold */
private IntList scaffoldLengths=new IntList();
/** scaffoldLengths[id] stores the number of kmers in that scaffold (excluding mutants) */
private IntList scaffoldKmers=new IntList();
/** scaffolds[id] stores the number of kmers in that scaffold */
private ArrayList<byte[]> scaffolds=new ArrayList<byte[]>();
/** Array of reference files from which to load kmers */
private ArrayList<String> ref=new ArrayList<String>();
/** Array of literal strings from which to load kmers */
private String[] literal=null;
/** Taxonomic tree */
private TaxTree tree;
/** Input reads */
private String in1=null, in2=null;
/** Input qual files */
private String qfin1=null, qfin2=null;
/** Output reads (matched and at least minlen) */
private String outm1=null, outm2=null;
/** Output reads (unmatched or shorter than minlen) */
private String outu1=null, outu2=null;
/** Per-sequence or per-reference output pattern */
private String outpattern=null;
/** Statistics output files */
private String outstats=null, outrpkm=null, outrefstats=null, outtax=null;
/** Barcode output files */
private String outbarcodes=null;
/** NCBI file mapping gi numbers to taxa IDs (gi_taxid_nucl.dmp) */
private String giTableFile=null;
/** NCBI file of taxonomy names (names.dmp) */
private String taxNameFile=null;
/** NCBI file of taxonomic tree (nodes.dmp) */
private String taxNodeFile=null;
/** File containing a serialized TaxTree */
private String taxTreeFile;
/** Store reference sequences */
private boolean storeRefBases=false;
/** Only look for fully-contained reference sequences */
private boolean processContainedRef=false;
/** Dump kmers here. */
private String dump=null;
/** Maximum input reads (or pairs) to process. Does not apply to references. -1 means unlimited. */
private long maxReads=-1;
/** Process this fraction of input reads. */
private float samplerate=1f;
/** Set samplerate seed to this value. */
private long sampleseed=-1;
/** Output reads in input order. May reduce speed. */
private final boolean ordered;
/** Make the middle base in a kmer a wildcard to improve sensitivity */
private boolean maskMiddle=true;
private int midMaskLen=0;
/** Store reference kmers with up to this many substitutions */
private int hammingDistance=0;
/** Search for query kmers with up to this many substitutions */
private int qHammingDistance=0;
/** Store reference kmers with up to this many edits (including indels) */
private int editDistance=0;
/** Always skip this many kmers between used kmers when hashing reference. */
private int refSkip=0;
private long taxNodeCountLimit=1;
private int taxNodeNumberLimit=-1;
private int taxNodeMinLevel=0;
private int taxNodeMaxLevel=TaxTree.parseLevel("domain");
private boolean trackBarcodes=false;
private BarcodeMappingStats barcodeData=null;
/*--------------------------------------------------------------*/
/*---------------- Statistics ----------------*/
/*--------------------------------------------------------------*/
long readsIn=0;
long fragsIn=0;
long basesIn=0;
long readsMatched=0;
long basesMatched=0;
long readsUnmatched=0;
long basesUnmatched=0;
long readsAmbig=0;//TODO
long readsQTrimmed=0;
long basesQTrimmed=0;
long readsFTrimmed=0;
long basesFTrimmed=0;
long readsQFiltered=0;
long basesQFiltered=0;
long refReads=0;
long refBases=0;
long refKmers=0;
long correctReads=0;
long incorrectReads=0;
long storedKmers=0;
/*--------------------------------------------------------------*/
/*---------------- Final Primitives ----------------*/
/*--------------------------------------------------------------*/
/** Correct errors via read overlap */
private final boolean ecc;
/** Look for reverse-complements as well as forward kmers. Default: true */
private final boolean rcomp;
/** Don't allow a read 'N' to match a reference 'A'.
* Reduces sensitivity when hdist>0 or edist>0. Default: false. */
private final boolean forbidNs;
/** AND bitmask with 0's at the middle base */
private final long middleMask;
/** Data structure to use. Default: ARRAYHF */
private final int tableType;
/** Normal kmer length */
private final int k;
/** k-1; used in some expressions */
private final int k2;
/** A read must share at least this many kmers to be considered a match. Default: 1 */
private final int minKmerHits;
/** A read must share at least this fraction of its kmers to be considered a match. Default: 0 */
private final float minKmerFraction;
/** Determines how to handle ambiguously-mapping reads */
private final int ambigMode;
/** Determines when to early-exit kmer matching */
private final int matchMode;
/** First and second must differ by more than this to be unambiguous. */
private final int clearzone;
/** First and second must differ by more than this fraction of read kmers to be unambiguous. */
private final float clearzoneFraction;
/** Calculate accuracy rate by parsing headers of synthetic reads */
private final boolean parsecustom;
/** Quality-trim the left side */
private final boolean qtrimLeft;
/** Quality-trim the right side */
private final boolean qtrimRight;
/** Trim bases at this quality or below. Default: 4 */
private final float trimq;
/** Error rate for trimming (derived from trimq) */
private final float trimE;
/** Throw away reads below this average quality after trimming. Default: 0 */
private final float minAvgQuality;
/** If positive, calculate average quality from the first X bases only. Default: 0 */
private final int minAvgQualityBases;
/** Throw away reads failing chastity filter (:Y: in read header) */
private final boolean chastityFilter;
/** Throw away reads containing more than this many Ns. Default: -1 (disabled) */
private final int maxNs;
/** Throw away reads containing without at least this many consecutive called bases. */
private int minConsecutiveBases=0;
/** Throw away reads shorter than this after trimming. Default: 10 */
private final int minReadLength;
/** Throw away reads longer than this after trimming. Default: Integer.MAX_VALUE */
private final int maxReadLength;
/** Toss reads shorter than this fraction of initial length, after trimming */
private final float minLenFraction;
/** Filter reads by whether or not they have matching kmers */
private final boolean kfilter;
/** Trim left bases of the read to this position (exclusive, 0-based) */
private final int forceTrimLeft;
/** Trim right bases of the read after this position (exclusive, 0-based) */
private final int forceTrimRight;
/** Trim this many rightmost bases of the read */
private final int forceTrimRight2;
/** Trim right bases of the read modulo this value.
* e.g. forceTrimModulo=50 would trim the last 3bp from a 153bp read. */
private final int forceTrimModulo;
/** If positive, only look for kmer matches in the leftmost X bases */
private int restrictLeft;
/** If positive, only look for kmer matches the rightmost X bases */
private int restrictRight;
/** Skip this many initial input reads */
private final long skipreads;
/** Pairs go to outbad if either of them is bad, as opposed to requiring both to be bad.
* Default: true. */
private final boolean removePairsIfEitherBad;
/** Print only statistics for scaffolds that matched at least one read
* Default: true. */
private final boolean printNonZeroOnly;
/** Rename reads to indicate what they matched.
* Default: false. */
private final boolean rename;
/** Use names of reference files instead of scaffolds.
* Default: false. */
private final boolean useRefNames;
/** Include the count when adding names */
private boolean addCount=true;
/** Only use the top hit when adding names */
private boolean topHitOnly=false;
/** Add 'unknown' to unmapped or ambiguous reads in rename mode */
private boolean addUnknown=false;
/** Fraction of kmers to skip, 0 to 15 out of 16 */
private final int speed;
/** Skip this many kmers when examining the read. Default 1.
* 1 means every kmer is used, 2 means every other, etc. */
private final int qSkip;
/** True if speed and qSkip are disabled. */
private final boolean noAccel;
/** Pick a single scaffold per read pair, rather than per read */
private final boolean keepPairsTogether;
/** Store match IDs in an IntList rather than int array */
private final boolean USE_COUNTVECTOR;
/** Gather taxonomic information */
private final boolean USE_TAXTREE;
private ConcurrentHashMap<String, StringCount> setMap;
/*--------------------------------------------------------------*/
/*---------------- Static Fields ----------------*/
/*--------------------------------------------------------------*/
/** Number of tables (and threads, during loading) */
private static final int WAYS=9; //123
/** Verbose messages */
public static final boolean verbose=false; //123
/** Number of reads output in the last run */
public static long lastReadsOut;
/** Print messages to this stream */
private static PrintStream outstream=System.err;
/** Permission to overwrite existing files */
public static boolean overwrite=true;
/** Permission to append to existing files */
public static boolean append=false;
/** Print speed statistics upon completion */
public static boolean showSpeed=true;
/** Display progress messages such as memory usage */
public static boolean DISPLAY_PROGRESS=true;
/** Number of ProcessThreads */
public static int THREADS=Shared.threads();
/** Indicates end of input stream */
private static final ArrayList<Read> POISON=new ArrayList<Read>(0);
/** Number of columns for statistics output, 3 or 5 */
public static int STATS_COLUMNS=5;
/** Release memory used by kmer storage after processing reads */
public static boolean RELEASE_TABLES=true;
/** Max value of hitCount array */
public static final int HITCOUNT_LEN=1000;
/** Make unambiguous copies of ref sequences with ambiguous bases */
public static boolean REPLICATE_AMBIGUOUS=false;
/** Write refstats in similar style to BBSplit */
public static boolean BBSPLIT_STYLE=false;
/** Rename duplicate scaffold names instead of crashing. */
public static boolean RENAME_DUPLICATES=true;
/** Create sets of amiguously-mapping gene targets on the fly */
public static boolean MAKE_GENE_SETS=false;
/** x&clearMasks[i] will clear base i */
private static final long[] clearMasks;
/** x|setMasks[i][j] will set base i to j */
private static final long[][] setMasks;
/** x&leftMasks[i] will clear all bases to the right of i (exclusive) */
private static final long[] leftMasks;
/** x&rightMasks[i] will clear all bases to the left of i (inclusive) */
private static final long[] rightMasks;
/** x|kMasks[i] will set the bit to the left of the leftmost base */
private static final long[] lengthMasks;
private static final int qhdistSizeLimit=10;
public static HashMap<String,String> RQC_MAP=null;
public static final int AMBIG_ALL=1, AMBIG_FIRST=2, AMBIG_TOSS=3, AMBIG_RANDOM=4;
public static final int MATCH_ALL=1, MATCH_FIRST=2, MATCH_UNIQUE=3;
/*--------------------------------------------------------------*/
/*---------------- Static Initializers ----------------*/
/*--------------------------------------------------------------*/
static{
clearMasks=new long[32];
leftMasks=new long[32];
rightMasks=new long[32];
lengthMasks=new long[32];
setMasks=new long[4][32];
for(int i=0; i<32; i++){
clearMasks[i]=~(3L<<(2*i));
}
for(int i=0; i<32; i++){
leftMasks[i]=((-1L)<<(2*i));
}
for(int i=0; i<32; i++){
rightMasks[i]=~((-1L)<<(2*i));
}
for(int i=0; i<32; i++){
lengthMasks[i]=((1L)<<(2*i));
}
for(int i=0; i<32; i++){
for(long j=0; j<4; j++){
setMasks[(int)j][i]=(j<<(2*i));
}
}
}
}
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