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package bloom;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import dna.AminoAcid;
import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import jgi.BBMerge;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import stream.SamLine;
import structures.ListNum;
import structures.LongList;
import tracker.ReadStats;
/**
* Wraps a BloomFilter to filter reads.
*
* @author Brian Bushnell
* @date April 23, 2018
*
*/
public class BloomFilterWrapper {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
* @param args Command line arguments
*/
public static void main(String[] args){
//Start a timer immediately upon code entrance.
Timer t=new Timer();
//Create an instance of this class
BloomFilterWrapper x=new BloomFilterWrapper(args);
//Run the object
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
/**
* Constructor.
* @param args Command line arguments
*/
public BloomFilterWrapper(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
boolean setInterleaved=false; //Whether interleaved was explicitly set.
//Set shared static variables
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Tools.max(Shared.threads()>1 ? 2 : 1, Shared.threads()>20 ? Shared.threads()/2 : Shared.threads()));
SamLine.SET_FROM_OK=true;
//Create a parser object
Parser parser=new Parser();
boolean setBits=false;
int k_=31;
int hashes_=2;
int minConsecutiveMatches_=3;
int bits_=1;
int minCount_=1;
boolean rcomp_=true;
boolean requireBothToMatch_=false;
boolean ecco_=false;
boolean merge_=false;
//Parse each argument
for(int i=0; i<args.length; i++){
String arg=args[i];
//Break arguments into their constituent parts, in the form of "a=b"
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
}else if(a.equals("ordered")){
ordered=Parse.parseBoolean(b);
}
else if(a.equalsIgnoreCase("k") || a.equalsIgnoreCase("bloomK") || a.equalsIgnoreCase("bloomFilterK")){
k_=Integer.parseInt(b);
}else if(a.equalsIgnoreCase("hashes") || a.equalsIgnoreCase("bloomHashes") || a.equalsIgnoreCase("bloomFilterHashes")){
hashes_=Integer.parseInt(b);
}else if(a.equalsIgnoreCase("minhits") || a.equalsIgnoreCase("bloomMinHits") || a.equalsIgnoreCase("bloomFilterMinHits")){
minConsecutiveMatches_=Integer.parseInt(b);
}else if(a.equals("rcomp")){
rcomp_=Parse.parseBoolean(b);
}else if(a.equals("bits")){
setBits=true;
bits_=Integer.parseInt(b);
assert(bits_>0);
}else if(a.equals("mincount")){
minCount_=Integer.parseInt(b);
// assert(minCount_>=1) : "mincount must be at least 1.";
// minCount_=Tools.max(1, minCount_);
}else if(a.equals("minprob")){
ReadCounter.minProb=Float.parseFloat(b);
}else if(a.equals("requireboth")){
requireBothToMatch_=Parse.parseBoolean(b);
}else if(a.equals("ecco")){
ecco_=Parse.parseBoolean(b);
}else if(a.equals("merge")){
merge_=Parse.parseBoolean(b);
}else if(a.equals("memfraction") || a.equals("memmult") || a.equals("memratio")){
memFraction=Float.parseFloat(b);
}else if(a.equals("cells")){
BloomFilter.OVERRIDE_CELLS=Parse.parseKMG(b);
}else if(a.equals("seed")){
KCountArray7MTA.setSeed(Parse.parseKMG(b));
}
else if(a.equals("serialin")){
serialIn=b;
}else if(a.equals("serialout")){
serialOut=b;
}
else if(a.equals("ref")){
boolean result=Tools.addFiles(b, ref);
assert(result) : "Could not add file(s): "+b;
}else if(a.equals("outm") || a.equals("outm1") || a.equals("outbad") || a.equals("outbad1") || a.equals("outmatch") || a.equals("outmatch1")){
outm1=b;
}else if(a.equals("outm2") || a.equals("outbad2") || a.equals("outmatch2")){
outm2=b;
}else if(a.equals("outu") || a.equals("outu1") || a.equals("out") || a.equals("out1")){
out1=b;
}else if(a.equals("outu2") || a.equals("out2")){
out2=b;
}else if(a.equals("outc") || a.equals("counts")){
outc=b;
}else if(a.equals("parse_flag_goes_here")){
long fake_variable=Parse.parseKMG(b);
//Set a variable here
}else if(parser.parse(arg, a, b)){//Parse standard flags in the parser
//do nothing
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
}
}
while(minCount_>0 && (1L<<bits_)-1<minCount_){bits_*=2;}
k=k_;
bits=bits_;
hashes=hashes_;
minConsecutiveMatches=minConsecutiveMatches_;
minCount=minCount_;
rcomp=rcomp_;
requireBothToMatch=requireBothToMatch_;
ecco=ecco_;
merge=merge_;
KmerCountAbstract.CANONICAL=rcomp; //rcomp, or true, perhaps... hmmm.
shift=bitsPerBase*k;
shift2=shift-bitsPerBase;
mask=(shift>63 ? -1L : ~((-1L)<<shift));
{//Process parser fields
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=ReadStats.overwrite=parser.overwrite;
append=ReadStats.append=parser.append;
setInterleaved=parser.setInterleaved;
in1=parser.in1;
in2=parser.in2;
qfin1=parser.qfin1;
qfin2=parser.qfin2;
qfout1=parser.qfout1;
qfout2=parser.qfout2;
extin=parser.extin;
extout=parser.extout;
}
//Do input file # replacement
if(in1!=null && in2==null && in1.indexOf('#')>-1 && !new File(in1).exists()){
in2=in1.replace("#", "2");
in1=in1.replace("#", "1");
}
//Do output file # replacement
if(out1!=null && out2==null && out1.indexOf('#')>-1){
out2=out1.replace("#", "2");
out1=out1.replace("#", "1");
}
//Do output file # replacement
if(outm1!=null && outm2==null && outm1.indexOf('#')>-1){
outm2=outm1.replace("#", "2");
outm1=outm1.replace("#", "1");
}
//Adjust interleaved detection based on the number of input files
if(in2!=null){
if(FASTQ.FORCE_INTERLEAVED){outstream.println("Reset INTERLEAVED to false because paired input files were specified.");}
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=false;
}
assert(FastaReadInputStream.settingsOK());
//Ensure there is an input file
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
//Adjust the number of threads for input file reading
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
//Ensure out2 is not set without out1
if(out1==null && out2!=null){throw new RuntimeException("Error - cannot define out2 without defining out1.");}
//Adjust interleaved settings based on number of output files
if(!setInterleaved){
assert(in1!=null && (out1!=null || out2==null)) : "\nin1="+in1+"\nin2="+in2+"\nout1="+out1+"\nout2="+out2+"\n";
if(in2!=null){ //If there are 2 input streams.
FASTQ.FORCE_INTERLEAVED=FASTQ.TEST_INTERLEAVED=false;
outstream.println("Set INTERLEAVED to "+FASTQ.FORCE_INTERLEAVED);
}else{ //There is one input stream.
if(out2!=null){
FASTQ.FORCE_INTERLEAVED=true;
FASTQ.TEST_INTERLEAVED=false;
outstream.println("Set INTERLEAVED to "+FASTQ.FORCE_INTERLEAVED);
}
}
}
//Ensure output files can be written
if(!Tools.testOutputFiles(overwrite, append, false, out1, out2, outm1, outm2)){
outstream.println((out1==null)+", "+(out2==null)+", "+out1+", "+out2);
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+out1+", "+out2+"\n");
}
//Ensure input files can be read
if(!Tools.testInputFiles(false, true, in1, in2)){
throw new RuntimeException("\nCan't read some input files.\n");
}
//Ensure that no file was specified multiple times
if(!Tools.testForDuplicateFiles(true, in1, in2, out1, out2, outm1, outm2)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
//Create output FileFormat objects
ffout1=FileFormat.testOutput(out1, FileFormat.FASTQ, extout, true, overwrite, append, ordered);
ffout2=FileFormat.testOutput(out2, FileFormat.FASTQ, extout, true, overwrite, append, ordered);
//Create output FileFormat objects
ffoutm1=FileFormat.testOutput(outm1, FileFormat.FASTQ, extout, true, overwrite, append, ordered);
ffoutm2=FileFormat.testOutput(outm2, FileFormat.FASTQ, extout, true, overwrite, append, ordered);
ffoutc=FileFormat.testOutput(outc, FileFormat.FASTA, extout, true, overwrite, append, ordered);
//Create input FileFormat objects
ffin1=FileFormat.testInput(in1, FileFormat.FASTA, extin, true, true);
ffin2=FileFormat.testInput(in2, FileFormat.FASTA, extin, true, true);
{
Timer t=new Timer(outstream, true);
if(serialIn==null){
filter=new BloomFilter(null, null, ref, k, k, bits, hashes, minConsecutiveMatches,
rcomp, ecco, merge, memFraction);
if(serialOut!=null){
ReadWrite.writeObjectInThread(filter, serialOut, true);
}
}else{
filter=ReadWrite.read(BloomFilter.class, serialIn, true);
}
t.stop("Filter creation: \t\t");
outstream.println(filter.filter.toShortString());
if(verbose){
outstream.println("Keys Counted: "+KmerCountAbstract.keysCounted);
outstream.println("Increments: "+KmerCountAbstract.increments);
}
}
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
void process(Timer t){
//Turn off read validation in the input threads to increase speed
final boolean vic=Read.VALIDATE_IN_CONSTRUCTOR;
Read.VALIDATE_IN_CONSTRUCTOR=Shared.threads()<4;
//Create a read input stream
final ConcurrentReadInputStream cris;
{
cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, ffin2, qfin1, qfin2);
cris.start(); //Start the stream
if(verbose){outstream.println("Started cris");}
}
boolean paired=cris.paired();
if(!ffin1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
//Optionally create read output streams
final ConcurrentReadOutputStream rosu, rosm, rosc;
if(ffout1!=null){
//Select output buffer size based on whether it needs to be ordered
final int buff=(ordered ? Tools.mid(16, 128, (Shared.threads()*2)/3) : 8);
//Notify user of output mode
if(cris.paired() && out2==null && (in1!=null && !ffin1.samOrBam() && !ffout1.samOrBam())){
outstream.println("Writing interleaved.");
}
rosu=ConcurrentReadOutputStream.getStream(ffout1, ffout2, qfout1, qfout2, buff, null, false);
rosu.start(); //Start the stream
}else{rosu=null;}
if(ffoutm1!=null){
//Select output buffer size based on whether it needs to be ordered
final int buff=(ordered ? Tools.mid(16, 128, (Shared.threads()*2)/3) : 8);
rosm=ConcurrentReadOutputStream.getStream(ffoutm1, ffoutm2, qfoutm1, qfoutm2, buff, null, false);
rosm.start(); //Start the stream
}else{rosm=null;}
if(ffoutc!=null){
//Select output buffer size based on whether it needs to be ordered
final int buff=(ordered ? Tools.mid(16, 128, (Shared.threads()*2)/3) : 8);
rosc=ConcurrentReadOutputStream.getStream(ffoutc, null, buff, null, false);
rosc.start(); //Start the stream
}else{rosc=null;}
//Reset counters
readsProcessed=readsOut=0;
basesProcessed=basesOut=0;
Timer t2=new Timer(outstream, true);
//Process the reads in separate threads
spawnThreads(cris, rosu, rosm, rosc);
if(verbose){outstream.println("Finished; closing streams.");}
//Write anything that was accumulated by ReadStats
errorState|=ReadStats.writeAll();
//Close the read streams
errorState|=ReadWrite.closeStreams(cris, rosu, rosm, rosc);
//Reset read validation
Read.VALIDATE_IN_CONSTRUCTOR=vic;
//Report timing and results
t.stop();
t2.stop("\nFiltering Time: \t\t");
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
{
long readsMatched=(readsProcessed-readsOut);
long basesMatched=(basesProcessed-basesOut);
double rpct=readsMatched*100.0/readsProcessed;
double bpct=basesMatched*100.0/basesProcessed;
String rstring=Tools.padKMB(readsMatched, 8);
String bstring=Tools.padKMB(basesMatched, 8);
StringBuilder sb=new StringBuilder();
sb.append("Reads Matched: ").append(rstring).append(Tools.format(" \t%.2f%%", rpct)).append('\n');
sb.append("Bases Matched: ").append(bstring).append(Tools.format(" \t%.2f%%", bpct));
outstream.println(sb);
}
//Throw an exception of there was an error in a thread
if(errorState){
throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
}
}
/** Spawn process threads */
private void spawnThreads(final ConcurrentReadInputStream cris,
final ConcurrentReadOutputStream rosu, final ConcurrentReadOutputStream rosm,
final ConcurrentReadOutputStream rosc){
//Do anything necessary prior to processing
//Determine how many threads may be used
final int threads=Shared.threads();
//Fill a list with ProcessThreads
ArrayList<ProcessThread> alpt=new ArrayList<ProcessThread>(threads);
for(int i=0; i<threads; i++){
alpt.add(new ProcessThread(cris, rosu, rosm, rosc, i));
}
//Start the threads
for(ProcessThread pt : alpt){
pt.start();
}
//Wait for completion of all threads
boolean success=true;
for(ProcessThread pt : alpt){
//Wait until this thread has terminated
while(pt.getState()!=Thread.State.TERMINATED){
try {
//Attempt a join operation
pt.join();
} catch (InterruptedException e) {
//Potentially handle this, if it is expected to occur
e.printStackTrace();
}
}
//Accumulate per-thread statistics
readsProcessed+=pt.readsProcessedT;
basesProcessed+=pt.basesProcessedT;
readsOut+=pt.readsOutT;
basesOut+=pt.basesOutT;
success&=pt.success;
}
//Track whether any threads failed
if(!success){errorState=true;}
//Do anything necessary after processing
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
/*--------------------------------------------------------------*/
/*---------------- Inner Classes ----------------*/
/*--------------------------------------------------------------*/
private class ProcessThread extends Thread {
//Constructor
ProcessThread(final ConcurrentReadInputStream cris_,
final ConcurrentReadOutputStream rosu_, final ConcurrentReadOutputStream rosm_,
final ConcurrentReadOutputStream rosc_, final int tid_){
cris=cris_;
rosu=rosu_;
rosm=rosm_;
rosc=rosc_;
tid=tid_;
}
//Called by start()
@Override
public void run(){
//Process the reads
processInner();
//Do anything necessary after processing
//Indicate successful exit status
success=true;
}
/** Iterate through the reads */
void processInner(){
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Grab the actual read list from the ListNum
ArrayList<Read> reads=(ln!=null ? ln.list : null);
//Check to ensure pairing is as expected
if(reads!=null && !reads.isEmpty()){
Read r=reads.get(0);
assert(ffin1.samOrBam() || (r.mate!=null)==cris.paired()); //Disabled due to non-static access
}
//As long as there is a nonempty read list...
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
// if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
ArrayList<Read> matched=new ArrayList<Read>(reads.size());
ArrayList<Read> counts=(rosc==null ? null : new ArrayList<Read>(reads.size()));
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
Read r1=reads.get(idx);
Read r2=r1.mate;
final String r2id=r1.mateId();
//Validate reads in worker threads
if(!r1.validated()){r1.validate(true);}
if(r2!=null && !r2.validated()){r2.validate(true);}
//Track the initial length for statistics
final int initialLength1=r1.length();
final int initialLength2=r1.mateLength();
//Increment counters
readsProcessedT+=r1.pairCount();
basesProcessedT+=initialLength1+initialLength2;
//Reads are processed in this block.
{
if((ecco || merge) && r1!=null && r2!=null){
if(merge){
final int insert=BBMerge.findOverlapStrict(r1, r2, false);
if(insert>0){
r2.reverseComplement();
r1=r1.joinRead(insert);
r2=null;
}
}else if(ecco){
BBMerge.findOverlapStrict(r1, r2, true);
}
}
if(rosc!=null){
addCounts(r1, counts);
addCounts(r2, counts);
}
final boolean match=matchesFilter(r1, r2);
if(match){
reads.set(idx, null);
matched.add(r1);
}else{
readsOutT+=r1.pairCount();
basesOutT+=r1.pairLength();
}
}
}
//Output reads to the output streams
if(rosu!=null){rosu.add(reads, ln.id);}
if(rosm!=null){rosm.add(matched, ln.id);}
if(rosc!=null){rosc.add(counts, ln.id);}
//Notify the input stream that the list was used
cris.returnList(ln);
// if(verbose){outstream.println("Returned a list.");} //Disabled due to non-static access
//Fetch a new list
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
/**
* Breaks a read into kmers and adds them, with their counts, to the new list.
* @param r Read to process
* @param counts Output list of kmers (bases) and counts (read name)
*/
private void addCounts(Read r, ArrayList<Read> counts){
if(r==null){return;}
final byte[] bases=r.bases;
long kmer=0;
long rkmer=0;
int len=0;
for(int i=0; i<bases.length; i++){
byte b=bases[i];
long x=AminoAcid.baseToNumber[b];
long x2=AminoAcid.baseToComplementNumber[b];
kmer=((kmer<<bitsPerBase)|x)&mask;
rkmer=((rkmer>>>bitsPerBase)|(x2<<shift2))&mask;
if(x<0){
len=0;
kmer=rkmer=0;
}else{
len++;
if(len>=k){
long key=rcomp ? Tools.max(kmer, rkmer) : kmer;
int count=filter.getCount(key);
if(count>=minCount){
String s=AminoAcid.kmerToString(key, k);
Read cread=new Read(s.getBytes(), null, ""+count, r.numericID);
counts.add(cread);
}
}
}
}
}
/**
* Process a read or a read pair.
* @param r1 Read 1
* @param r2 Read 2 (may be null)
* @return True if the reads should be kept (meaning they don't match the filter), false if they should be discarded.
*/
boolean matchesFilter(final Read r1, final Read r2){
if(r2!=null && requireBothToMatch){
// assert(false) : minCount+", "+filter.matches(r1, buffer, minCount)+", "+filter.matches(r2, buffer, minCount)+" "+filter.matchesEither(r1, r2, buffer, minCount);
return filter.matches(r1, buffer, minCount) && filter.matches(r2, buffer, minCount);
}else{
// assert(false) : minCount+", "+filter.matches(r1, buffer, minCount)+", "+filter.matches(r2, buffer, minCount)+" "+filter.matchesEither(r1, r2, buffer, minCount)+"\n"+r1+"\n"+r2;
return filter.matchesEither(r1, r2, buffer, minCount);
}
}
private final LongList buffer=minConsecutiveMatches>1 ? new LongList(150) : null;
/** Number of reads processed by this thread */
protected long readsProcessedT=0;
/** Number of bases processed by this thread */
protected long basesProcessedT=0;
/** Number of reads retained by this thread */
protected long readsOutT=0;
/** Number of bases retained by this thread */
protected long basesOutT=0;
/** True only if this thread has completed successfully */
boolean success=false;
/** Shared input stream */
private final ConcurrentReadInputStream cris;
/** Shared output stream */
private final ConcurrentReadOutputStream rosu;
/** Matched output stream */
private final ConcurrentReadOutputStream rosm;
/** Count output stream */
private final ConcurrentReadOutputStream rosc;
/** Thread ID */
final int tid;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
private ArrayList<String> ref=new ArrayList<String>();
/** Primary input file path */
private String in1=null;
/** Secondary input file path */
private String in2=null;
private String qfin1=null;
private String qfin2=null;
/** Primary output file path */
private String out1=null;
/** Secondary output file path */
private String out2=null;
/** Output for kmer counts */
private String outc=null;
private String qfout1=null;
private String qfout2=null;
/** Output file path for matching reads */
private String outm1=null;
/** Secondary output file path for matching reads */
private String outm2=null;
private String qfoutm1=null;
private String qfoutm2=null;
/** Override input file extension */
private String extin=null;
/** Override output file extension */
private String extout=null;
private String serialIn=null;
private String serialOut=null;
/*--------------------------------------------------------------*/
/** Number of reads processed */
protected long readsProcessed=0;
/** Number of bases processed */
protected long basesProcessed=0;
/** Number of reads retained */
protected long readsOut=0;
/** Number of bases retained */
protected long basesOut=0;
/** Quit after processing this many input reads; -1 means no limit */
private long maxReads=-1;
/*--------------------------------------------------------------*/
/*---------------- Final Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file */
final FileFormat ffin1;
/** Secondary input file */
final FileFormat ffin2;
/** Primary output file */
private final FileFormat ffout1;
/** Secondary output file */
private final FileFormat ffout2;
/** Primary output file for matching reads */
private final FileFormat ffoutm1;
/** Secondary output file for matching reads */
private final FileFormat ffoutm2;
/** Output for kmers and counts */
private final FileFormat ffoutc;
final BloomFilter filter;
final int k;
final int hashes;
final int bits;
final int minConsecutiveMatches;
final boolean rcomp;
final boolean requireBothToMatch;
final boolean ecco;
final boolean merge;
final int minCount;
float memFraction=1.0f;
final int bitsPerBase=BloomFilter.bitsPerBase;
final int shift;
final int shift2;
final long mask;
/*--------------------------------------------------------------*/
/*---------------- Common Fields ----------------*/
/*--------------------------------------------------------------*/
/** Print status messages to this output stream */
private PrintStream outstream=System.err;
/** Print verbose messages */
public static boolean verbose=false;
/** True if an error was encountered */
public boolean errorState=false;
/** Overwrite existing output files */
private boolean overwrite=true;
/** Append to existing output files */
private boolean append=false;
/** Reads are output in input order */
private boolean ordered=false;
}
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