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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.BitSet;
import java.util.HashMap;
import java.util.HashSet;
import java.util.Locale;
import java.util.Map.Entry;
import java.util.regex.Pattern;
import dna.AminoAcid;
import fileIO.ByteFile;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import fileIO.TextFile;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.ReadStats;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.BufferedMultiCros;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.MultiCros2;
import stream.MultiCros3;
import stream.Read;
import stream.SamLine;
import structures.ByteBuilder;
import structures.ListNum;
/**
* This class is designed to handle very large numbers of output files
* with a fixed number of file handles. An example use case is demultiplexing
* Illumina Novaseq runs.
*
* @author Brian Bushnell
* @date May 1, 2019
*
*/
public class DemuxByName2 {
/** Code entrance from the command line */
public static void main(String[] args){
//Capture values of static variables that might be modified in case this is called by another class.
final int oldCap=Shared.numBuffers(), oldZipThreads=ReadWrite.MAX_ZIP_THREADS, oldZl=ReadWrite.ZIPLEVEL;
//External compressor selection
final boolean oldPigz=ReadWrite.USE_PIGZ, oldUnpigz=ReadWrite.USE_UNPIGZ;
final boolean oldBgzip=ReadWrite.USE_BGZIP, oldPreferBgzip=ReadWrite.PREFER_BGZIP;
//Preserve quality scores even for reads with with incorrect quality scores
final boolean oldCQ=Read.CHANGE_QUALITY;
//Create and start a timer, for statistics
Timer t=new Timer();
//Create the demultiplexer object
DemuxByName2 demultiplexer=new DemuxByName2(args);
//Process all data
demultiplexer.process(t);
//Restore values of static variables.
Shared.setBuffers(oldCap);
ReadWrite.ZIPLEVEL=oldZl;
ReadWrite.USE_PIGZ=oldPigz;
ReadWrite.USE_BGZIP=oldBgzip;
ReadWrite.PREFER_BGZIP=oldPreferBgzip;
ReadWrite.USE_UNPIGZ=oldUnpigz;
ReadWrite.MAX_ZIP_THREADS=oldZipThreads;
Read.CHANGE_QUALITY=oldCQ;
//Close the print stream if it was redirected
Shared.closeStream(demultiplexer.outstream);
}
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Constructor.
* @param args Command line arguments
*/
public DemuxByName2(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
//Set some static variables
Shared.capBuffers(4);
ReadWrite.USE_PIGZ=true;
ReadWrite.USE_UNPIGZ=true;
ReadWrite.MAX_ZIP_THREADS=Shared.threads();
SamLine.SET_FROM_OK=true;
//Reduce default compression level to increase speed
//This automatically gets bumped up to 4 if pigz/bgzip is detected
ReadWrite.ZIPLEVEL=2;
Read.CHANGE_QUALITY=false;
{//Argument-parsing block
//Parse arguments
final Parser parser=parse(args);
//Process parser fields
//This is in the constructor rather than a function to allow final fields
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=ReadStats.overwrite=parser.overwrite;
append=ReadStats.append=parser.append;
setInterleaved=parser.setInterleaved;
trackCardinality=parser.loglog;
in1=parser.in1;
in2=parser.in2;
qfin1=parser.qfin1;
qfin2=parser.qfin2;
out1=parser.out1;
out2=parser.out2;
qfout1=parser.qfout1;
qfout2=parser.qfout2;
extin=parser.extin;
extout=parser.extout;
}
//Ensure the input and output files are specified correctly
validate();
//Create FileFormat opjects for input files
ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
ffin2=FileFormat.testInput(in2, FileFormat.FASTQ, extin, true, true);
//If the input and output are in sam/bam format, print the sam header
if(ffin1!=null && out1!=null && ffin1.samOrBam()){
String ext=ReadWrite.rawExtension(out1);
useSharedHeader=FileFormat.isSamOrBam(ext);
}
//Set up data structures
handleModeAndNames();
}
/**
* Parse command-line arguments
* @param args Command-line arguments
* @return A Parser object with fields filled from the parameters
*/
private Parser parse(final String[] args){
//Handles parsing of common flags
Parser parser=new Parser();
parser.overwrite=overwrite;
//Handles parsing of flags specific to this program
for(int i=0; i<args.length; i++){//Parsing loop
String arg=args[i];
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);
BufferedMultiCros.verbose=verbose;
}else if(a.equals("names") || a.equals("name") || a.equals("affixes")){
if(b!=null){
String[] x=b.split(",");
for(String s : x){
names.put(s, s);
}
}
}else if(a.equals("threaded")){
threaded=Parse.parseBoolean(b);
}else if(a.equals("chrom") || a.equals("chrommode") || a.equals("scaffold")){
if(Parse.parseBoolean(b)){mode=CHROM_MODE;}
}else if(a.equals("substringmode") || a.equals("substring")){
if(Parse.parseBoolean(b)){mode=SUBSTRING_MODE;}
}else if(a.equals("barcode") || a.equals("barcodemode") || a.equals("index")){
if(Parse.parseBoolean(b)){mode=BARCODE_MODE;}
}else if(a.equals("perheader") || a.equals("persequence") || a.equals("header") || a.equals("headermode")){
if(Parse.parseBoolean(b)){mode=HEADER_MODE;}
}else if(a.equals("affix") || a.equals("affixmode")){
if(Parse.parseBoolean(b)){mode=AFFIX_MODE;}
}else if(a.equals("delimiter")){
delimiter=Parse.parseSymbol(b);
if(delimiter!=null){mode=DELIMITER_MODE;}
}else if(a.equals("prefixmode") || a.equals("prefix") || a.equals("pm")){
prefixMode=Parse.parseBoolean(b);
}else if(a.equals("suffixmode") || a.equals("suffix") || a.equals("sm")){
prefixMode=!Parse.parseBoolean(b);
}else if(a.equals("column")){
column=Integer.parseInt(b);
assert(column>0 || column==-1) : "Column is 1-based; must be 1+ or else -1 to disable.";
column--;
}else if(a.equalsIgnoreCase("length") || a.equalsIgnoreCase("len") || a.equalsIgnoreCase("affixlength") || a.equalsIgnoreCase("affixlen")){
fixedLength=Integer.parseInt(b); //also set affix mode, perhaps
}else if(a.equalsIgnoreCase("stats") || a.equalsIgnoreCase("report") || a.equalsIgnoreCase("results")){
stats=b;
}else if(a.equalsIgnoreCase("minreadstodump") || a.equalsIgnoreCase("minreads")){
minReadsToDump=Parse.parseKMG(b);
}else if(a.equalsIgnoreCase("mcrostype")){
mcrosType=Integer.parseInt(b);
}else if(a.equalsIgnoreCase("streams") || a.equalsIgnoreCase("maxstreams")){
maxStreams=Integer.parseInt(b);
}else if(a.equalsIgnoreCase("readsperbuffer") || a.equalsIgnoreCase("rpb")){
readsPerBuffer=Parse.parseIntKMG(b);
}else if(a.equalsIgnoreCase("bytesPerBuffer") || a.equalsIgnoreCase("bpb")){
bytesPerBuffer=Parse.parseIntKMG(b);
}else if(a.equals("hdist") || a.equals("hamming") || a.equals("hammingdistance")){
hdist=Integer.parseInt(b);
}else if(a.equals("outu") || a.equals("outu1")){
outu1=b;
}else if(a.equals("outu2")){
outu2=b;
}else if(a.equals("pattern")){
parser.out1=b;
}else if(a.equals("remap") || a.equals("replace")){
remap=Parse.parseRemap(b);
}
//Old version, allowed more complicated replacements
// else if(a.equals("replace")){
// if(b==null){
// replaceA=replaceB=null;
// }else{
// if(b.length()==2){
// replaceA=b.substring(0, 1);
// replaceB=b.substring(1);
// }else{
// String[] pair=b.split(",");
// replaceA=Parse.parseSymbol(pair[0]);
// replaceB=Parse.parseSymbol(pair[1]);
// }
// }
// }
else if(parser.parse(arg, a, b)){
//Flag was captured by the parser; do nothing
}else if(parser.in1==null && i==0 && !arg.contains("=") && (arg.toLowerCase().startsWith("stdin") || new File(arg).exists())){
parser.in1=arg;
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
}
}
return parser;
}
/**
* Ensure input and output files are specified correctly,
* and that interleaved mode is consistent with the number of input files.
*/
private void validate(){
//Validate patterns
assert(out1==null || out1.contains("%")) : "Output filename must contain '%' symbol, which will be replaced by affix.";
assert(out2==null || out2.contains("%")) : "Output filename must contain '%' symbol, which will be replaced by affix.";
assert(qfout1==null || qfout1.contains("%")) : "Output filename must contain '%' symbol, which will be replaced by affix.";
assert(qfout2==null || qfout2.contains("%")) : "Output filename must contain '%' symbol, which will be replaced by affix.";
//Perform # replacement for twin files
if(in1!=null && in2==null && in1.indexOf('#')>-1 && !new File(in1).exists()){
in2=in1.replace("#", "2");
in1=in1.replace("#", "1");
}
if(out1!=null && out2==null && out1.indexOf('#')>-1){
out2=out1.replace("#", "2");
out1=out1.replace("#", "1");
}
if(outu1!=null && outu2==null && outu1.indexOf('#')>-1){
outu2=outu1.replace("#", "2");
outu1=outu1.replace("#", "1");
}
//Disable interleaving if in2 is specified
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;
}
{//Perform various file validation
assert(FastaReadInputStream.settingsOK());
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
if(out1==null && out2!=null){throw new RuntimeException("Error - cannot define out2 without defining out1.");}
//ByteFile2 is multithreaded; only use it if there are plenty of threads
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
//Do some interleaving-detection logic if unspecified
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);
}
}
}
//If the user explicitly used the word "null", change it to null.
if(out1!=null && out1.equalsIgnoreCase("null")){out1=null;}
if(out2!=null && out2.equalsIgnoreCase("null")){out2=null;}
//Ensure output files have no duplicates and can be written
//Unfortunately, this does not prevent patterns expanding into a matching string
if(!Tools.testOutputFiles(overwrite, append, false, outu1, outu2, stats)){
outstream.println((out1==null)+", "+(out2==null)+", "+out1+", "+out2+", "+outu1+", "+outu2+", "+stats);
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+out1+", "+out2+", "+outu1+", "+outu2+", "+stats+"\n");
}
//This does not prevent expansion into a matching string
assert(!out1.equalsIgnoreCase(in1) && !out1.equalsIgnoreCase(in1)) : "Input file and output file have same name.";
assert(out2==null || (!out2.equalsIgnoreCase(in1) && !out2.equalsIgnoreCase(in2))) : "out1 and out2 have same name.";
}
}
/**
* Perform necessary setup of variables and data structures
* related to the processing mode and the names table.
*/
private void handleModeAndNames(){
//Handle names
if(names.isEmpty()){
names=null;
}else{
{//Process names, because they can either be literals or filenames at this point
String[] keys=names.keySet().toArray(new String[names.size()]);
names.clear();
for(String key : keys){//Process either literal names or files of names
File f=new File(key);
if(f.exists() && f.isFile()){//Read the file
TextFile tf=new TextFile(key);
String[] lines=tf.toStringLines();
for(String line : lines){names.put(line, line);}//Add each line as a name
}else{names.put(key, key);}//Re-add each key as a literal name
}
}
}
//Handle affix lengths
if(mode==AFFIX_MODE && names!=null){
{//Find the lengths of all names to fill the lengthArray array
BitSet bs=new BitSet();//A set of observed affix lengths
//Add the fixed affix length, if present
if(fixedLength>0){bs.set(fixedLength);}
//Add the lengths of everything in the names set
for(String s : names.keySet()){bs.set(s.length());}
//Put the set members into an array for faster access
lengthArray=new int[bs.cardinality()];
for(int i=0, bit=-1; i<lengthArray.length; i++){
bit=bs.nextSetBit(bit+1);
lengthArray[i]=bit;
}
//Sort and reverse so that the lengths are descending
Arrays.sort(lengthArray);
Tools.reverseInPlace(lengthArray);
}
assert((lengthArray.length>0 && lengthArray[0]>0)) :
"Must include at least one name, an affix length, or a delimiter.";
//If there are more than one length, disable the fixedLength mode
if(lengthArray!=null && lengthArray.length>1){fixedLength=-1;}
else if(lengthArray.length==1){
assert(fixedLength<1 || fixedLength==lengthArray[0]) :
"\nLength flag ("+fixedLength+") does not match detected name length of "+lengthArray[0]+
"\nPlease omit the length flag or set it correctly.\n";
fixedLength=lengthArray[0];
lengthArray=null;
}
}
//For length-1 delimiters, use a char instead of a String
assert(column<0 || delimiter!=null) : "Column may not be set if there is no delimiter.";
if(delimiter!=null){
if(delimiter.length()==1){
delimiterChar=delimiter.charAt(0);
}else if(delimiter.length()==2 && delimiter.charAt(0)=='\\'){
delimiterChar=delimiter.charAt(1);
}else{
delimiterChar=0;
}
delimiterPattern=Pattern.compile(delimiter);
}
//Populate names table with mutants for hamming distance
if(hdist>0 && names.size()>0){
@SuppressWarnings("unused")
int numMutants=mutate(names, hdist, outstream);
}
//Populate name list with keys from the name map
if(names==null){
nameList=null;
}else{
for(Entry<String, String> e : names.entrySet()){
nameList.add(e.getKey());
}
}
//Ensure there are names in substring mode
if(mode==SUBSTRING_MODE){
assert(nameList!=null && !nameList.isEmpty()) : "Empty names list is not allowed in substring mode.";
}
//Perform symbol replacements in output filenames
if(remap!=null && names!=null && !names.isEmpty()){
for(String key : nameList){
String old=names.get(key);
String value=Tools.remap(remap, old);
if(!value.equals(old)){names.put(key, value);}
}
}
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* Primary method.
* Starts and stops I/O streams, processes the data, and prints results.
* @param t A timer that was already started.
*/
void process(Timer t){
//Create stream for input reads
final ConcurrentReadInputStream cris=makeInputStream();
//Create streams for output reads other than unmatched
final BufferedMultiCros mcros=makeMatchedOutputStream(cris.paired());
//Create stream for unmatched output reads
final ConcurrentReadOutputStream rosu=makeUnmatchedOutputStream();
//Streams are set up, so process the reads
processInner(cris, mcros, rosu);
//At this point processing has finished.
//Close streams
cleanup(cris, mcros, rosu);
//Report statistics to file
if(stats!=null){printReport(mcros);}
//Stop the timer
t.stop();
//Print results
printResultsToScreen(t);
if(errorState){//Exit with a message and error code
throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
}
}
/** Create and start the input stream (cris). */
private ConcurrentReadInputStream makeInputStream(){
//Stream for input reads
final ConcurrentReadInputStream cris=ConcurrentReadInputStream.getReadInputStream(maxReads, useSharedHeader, ffin1, ffin2, qfin1, qfin2);
if(verbose){outstream.println("Started cris");}
cris.start();
//Report whether the input is being processed as paired.
final boolean paired=cris.paired();
if(!ffin1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
return cris;
}
/** Create and start the normal output streams (mcros). */
private BufferedMultiCros makeMatchedOutputStream(boolean pairedInput){
//Streams for output reads other than unmatched
final BufferedMultiCros mcros;
if(out1!=null){
if(mcrosType==2){//Slow, synchronous mcros type
mcros=new MultiCros2(out1, out2, overwrite, append, true, useSharedHeader, FileFormat.FASTQ, threaded);
}else if(mcrosType==3){//Faster, asynchronous type
mcros=new MultiCros3(out1, out2, overwrite, append, true, useSharedHeader, FileFormat.FASTQ, threaded, maxStreams);
}else{
throw new RuntimeException("Bad mcrosType: "+mcrosType);
}
//Set mcros fields not available in constructor
mcros.readsPerBuffer=readsPerBuffer;
mcros.bytesPerBuffer=bytesPerBuffer;
mcros.minReadsToDump=minReadsToDump;
mcros.trackCardinality=trackCardinality;
//Report output format
if(pairedInput && out2==null && (in1==null || !ffin1.samOrBam())){
outstream.println("Writing interleaved.");
}
//Start the mcros thread if necessary
if(threaded){mcros.start();}
}else{
mcros=null;
}
return mcros;
}
/** Create and start the unmatched output stream (rosu). */
private ConcurrentReadOutputStream makeUnmatchedOutputStream(){
//Stream for unmatched output reads
final ConcurrentReadOutputStream rosu;
if(outu1!=null){
//Number of output buffers; does not need to be high since access is single-threaded
final int buff=4;
FileFormat ffout1=FileFormat.testOutput(outu1, FileFormat.FASTQ, extout, true, overwrite, append, false);
FileFormat ffout2=(outu2==null ? null : FileFormat.testOutput(outu2, FileFormat.FASTQ, extout, true, overwrite, append, false));
rosu=ConcurrentReadOutputStream.getStream(ffout1, ffout2, null, null, buff, null, true);
rosu.start();
}else{
rosu=null;
}
return rosu;
}
/**
* Process all reads.
* @param cris Input stream (required).
* @param mcros Matched read output stream (optional).
* @param rosu Unmatched read output stream (optional).
*/
private void processInner(final ConcurrentReadInputStream cris, final BufferedMultiCros mcros, final ConcurrentReadOutputStream rosu) {
//Fetch the first list
ListNum<Read> ln=cris.nextList();
ArrayList<Read> reads=(ln!=null ? ln.list : null);
//Ensure pairing seems correct
if(reads!=null && !reads.isEmpty()){
Read r=reads.get(0);
assert((ffin1==null || ffin1.samOrBam()) || (r.mate!=null)==cris.paired());
}
//While there are more reads to process...
while(ln!=null && reads!=null && reads.size()>0){//ln!=null prevents a compiler potential null access warning
//List to accumulate unmatched reads
ArrayList<Read> unmatched=new ArrayList<Read>();
//For each read...
for(Read r : reads){
//Get the target file identifier
String name=getValue(r);
final int pairCount=r.pairCount(), pairLen=r.pairLength();
if(name!=null){
//Set the name so that the mcros will send it to the right place
r.obj=name;
readsOut+=pairCount;
basesOut+=pairLen;
}else{
//Send it to unmatched
unmatched.add(r);
readsUnmatched+=pairCount;
basesUnmatched+=pairLen;
}
readsProcessed+=pairCount;
basesProcessed+=pairLen;
}
if(rosu!=null){rosu.add(unmatched, ln.id);}//Send unmatched reads to rosu
if(mcros!=null){mcros.add(reads);}//Send matched reads to mcros
//Notify the input stream that the list has been processed
cris.returnList(ln);
//Fetch a new list
ln=cris.nextList();
reads=(ln!=null ? ln.list : null);
}
//Notify the input stream that this thread is done processing
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
/**
* Finish writing and close streams.
* @param cris Input stream (required).
* @param mcros Matched read output stream (optional).
* @param rosu Unmatched read output stream (optional).
*/
private void cleanup(final ConcurrentReadInputStream cris, final BufferedMultiCros mcros, final ConcurrentReadOutputStream rosu){
//Shut down mcros
if(mcros!=null){
mcros.close();
errorState|=mcros.errorState();
}
if(minReadsToDump>0 && mcros!=null){
//Dump the residual reads into the unmatched file
mcros.dumpResidual(rosu);
//Adjust statistics to reflect that residuals did not get demultiplexed
readsOut-=mcros.residualReads;
basesOut-=mcros.residualBases;
}
//errorState|=ReadStats.writeAll(); //Currently unused; allows tracking statistics.
//Shut down normal streams
errorState|=ReadWrite.closeStreams(cris, rosu);
}
/**
* @param t A Timer that has already been stopped
*/
private void printResultsToScreen(Timer t) {
//Report statistics to screen
double rpnano=readsProcessed/(double)(t.elapsed);
double bpnano=basesProcessed/(double)(t.elapsed);
outstream.println("Time: "+t);
outstream.println("Reads Processed: "+readsProcessed+" \t"+String.format(Locale.ROOT, "%.2fk reads/sec", rpnano*1000000));
outstream.println("Bases Processed: "+basesProcessed+" \t"+String.format(Locale.ROOT, "%.2fm bases/sec", bpnano*1000));
outstream.println("Reads Out: "+readsOut);
outstream.println("Bases Out: "+basesOut);
}
/*--------------------------------------------------------------*/
/*---------------- Helper Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* Generate mutants for all names according to the hamming distance
* @param names Map of names to mutate
* @param hdist Hamming distance
* @param outstream Report results here
* @return Number of mutants added (net)
*/
private static int mutate(HashMap<String, String> names, int hdist, PrintStream outstream){
//Put the names in a list for iteration, to prevent concurrent modification of the table
ArrayList<String> list=new ArrayList<String>(names.size());
list.addAll(names.values());
//Number of mutants generated
int mutants=0;
//Set of colliding keys to be removed
final HashSet<String> collisions=new HashSet<String>();
//Generate the mutants
for(String name : list){
mutants+=addMutants(name.getBytes(), name, hdist, names, collisions);
}
outstream.println("Added "+mutants+" mutants to the initial "+list.size()+" names.");
//Remove original keys from collisions so that it only contains mutants
collisions.removeAll(list);
int removed=0;
if(!collisions.isEmpty()){//Remove colliding mutants from the name table
for(String key : collisions) {
names.remove(key);
removed++;
}
outstream.println("Removed "+removed+" collisions due to ambiguity.");
}
final int netChange=mutants-removed;
return netChange;
}
/**
* Generate mutants for one name, recursively
* @param keyArray Current key (name) as a byte array, to be modified
* @param value Name to which this key should map
* @param hdist Remaining Hamming distance
* @param names Names table to insert the mutants
* @param collisions Set of keys mapping to multiple values
* @return Number of mutants added
*/
private static int addMutants(final byte[] keyArray, final String value, final int hdist, final HashMap<String, String> names, final HashSet<String> collisions){
assert(hdist>0);
int added=0;
//For each position in the array, change it to all possible values
for(int i=0; i<keyArray.length; i++){
final byte old=keyArray[i];
if(AminoAcid.isACGTN(old)){//Only mutate bases
for(byte b : symbols){
if(b!=old){
keyArray[i]=b;
{
String key=new String(keyArray);
String oldValue=names.get(key);
if(oldValue==null){
names.put(key, value);
added++;
// outstream.println("Added "+key+"->"+value+"; oldValue="+oldValue);
}else if(!oldValue.equals(value)){
// assert(value.equals(names.get(s))) : "Collision between "+value+" and "+names.get(s)+" for mutant "+s;
collisions.add(key);
// outstream.println("Collision for "+key+"->"+value+"; oldValue="+oldValue);
}
}
if(hdist>1){
//Recur on the mutant, with a reduced Hamming distance
added+=addMutants(keyArray, value, hdist-1, names, collisions);
}
}
}
//Restore the original value before moving to the next position
keyArray[i]=old;
}
}
return added;
}
/** Returns the value this read maps to; meaning, the variable part of the output filename */
private String getValue(Read r){
final String key=getKey(r);
if(verbose){System.err.println("Got key "+key);}
if(key==null){return null;}
//Return the value this key maps to
if(names!=null){
if(verbose){System.err.println("Got value "+names.get(key));}
return names.get(key);
}
//Return the value generated by remapping the key's symbols, if needed
final String value=(remap==null ? key : Tools.remap(remap, key));
if(verbose){System.err.println("Returning value "+value);}
return value;
}
/** Generates a key from the read header */
private String getKey(Read r){
final String id=r.id;
final int idlen=id.length();
final String key;
//Simplest case; every header is its own key
if(mode==HEADER_MODE){return id;}
//Parse the barcode
if(mode==BARCODE_MODE){return r.barcode(true);}
//Use mapping information
if(mode==CHROM_MODE){return r.rnameS();}
//Return a prefix or suffix
if(mode==AFFIX_MODE){
if(fixedLength>0){//If there is a single fixed affix length, return the affix
key=(id.length()<=fixedLength ? id : prefixMode ? id.substring(0, fixedLength) : id.substring(idlen-fixedLength));
return key;
}else{//Make a substring of each possible length and look it up in the hashmap
for(int affixLen : lengthArray){
final String sub=idlen>=affixLen ? prefixMode ? id.substring(0, affixLen) : id.substring(idlen-affixLen) : id;
if(names.containsKey(sub)){return sub;}
}
return null;
}
}
//Return a delimited substring
if(mode==DELIMITER_MODE){
if(column>-1){//Use a split operation, which is slow
// String[] split=id.split(delimiter);
String[] split=delimiterPattern.split(id);//Faster; uses a precompiled pattern
assert(split.length>1) : "Delimiter '"+delimiter+"' was not found in name '"+id+"'";
int col=Tools.min(column, split.length-1);
key=split[col];
if(col!=column && !warned){
outstream.println("*** WARNING! ***\n"
+ "Only "+(col+1)+" columns for record "+id+"\n"
+ "Further warnings will be suppressed.\n");
warned=true;
assert(errorState=true); //Change error state to true if assertions are enabled.
}
}else if(prefixMode){//Use a delimited prefix, which is fast
//Find the first index of the delimiter, using the char when possible
int idx=(delimiterChar>0 ? id.indexOf(delimiterChar) : id.indexOf(delimiter));
assert(idx>=0) : "Delimiter\r\n" +
" System.err.println(\"a\"); '"+delimiter+"' was not found in name '"+id+"'";
key=id.substring(0, idx);
}else{//Use a delimited suffix, which is fast
//Find the last index of the delimiter, using the char when possible
int idx=(delimiterChar>0 ? id.lastIndexOf(delimiterChar) : id.lastIndexOf(delimiter));
assert(idx>=0) : "Delimiter '"+delimiter+"' was not found in name '"+id+"'";
key=id.substring(idx+delimiter.length());
}
return key;
}
//Substring mode requires brute-force matching and is very slow
if(mode==SUBSTRING_MODE){
if(nameList.size()>0){
for(String s : nameList){
if(id.contains(s)){return s;}
}
return null;
}
}
throw new RuntimeException("No mode is being used: "+mode); //Should be unreachable
}
/**
* Print statistics about demultiplexing to a file.
* @param mcros Output stream, after processing is completely finished.
*/
void printReport(BufferedMultiCros mcros){
if(stats==null){return;}
//Make a writer for the stats file
ByteStreamWriter bsw=new ByteStreamWriter(stats, overwrite, append, true);
bsw.start();
{
ByteBuilder bb=new ByteBuilder();
//Print the header
bb.append("#ReadsIn\t").append(readsProcessed).nl();
bb.append("#BasesIn\t").append(basesProcessed).nl();
bb.append("#ReadsOut\t").append(readsOut).nl();
bb.append("#BasesOut\t").append(basesOut).nl();
bb.append("#Name\tReads\tBases"+(trackCardinality ? "\tCardinality" : "")+"\n");
//Print results of unmatched reads, which is not captured by the mcros
if(names!=null && !names.isEmpty()){
bb.append("Unmatched\t").append(readsUnmatched).tab().append(basesUnmatched).nl();
}
bsw.print(bb);
}
if(mcros!=null){//Print results from the mcros
ByteBuilder bb=mcros.report();
bsw.print(bb);
}
//Finish writing
bsw.poisonAndWait();
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file */
private String in1=null;
/** Optional input file for read 2 of paired reads in twin files */
private String in2=null;
/** Input qual file 1 */
private String qfin1=null;
/** Input qual file 2 */
private String qfin2=null;
/** Output file pattern 1 */
private String out1=null;
/** Output file pattern 2 */
private String out2=null;
/** Output qual file 1 */
private String qfout1=null;
/** Output qual file 2 */
private String qfout2=null;
/** Unmatched read output file 1 */
private String outu1=null;
/** Unmatched read output file 2 */
private String outu2=null;
/** File extension override for input files */
private String extin=null;
/** File extension override for output files */
private String extout=null;
/*--------------------------------------------------------------*/
/** Primary input file */
private final FileFormat ffin1;
/** Read 2 input file */
private final FileFormat ffin2;
/*--------------------------------------------------------------*/
/** Input reads */
long readsProcessed=0;
long basesProcessed=0;
/** Demultiplexed output reads */
long readsOut=0;
long basesOut=0;
/** Output reads that did not get demultiplexed */
long readsUnmatched=0;
long basesUnmatched=0;
/** Stop after this many input reads */
private long maxReads=-1;
/** File to print number of reads sent to each output file */
private String stats=null;
/** For splitting headers on a symbol */
private String delimiter=null;
/** For splitting headers faster if the delimiter is just one character */
private char delimiterChar=0;
/** Precompiled pattern matching the delimiter */
private Pattern delimiterPattern=null;
/** For replacing symbols in the filename */
private byte[] remap=null;
/** If there is a delimiter, use this column after splitting.
* Column is 1-based. */
private int column=-1;
/** Use the prefix of a header. If false, use the suffix.
* This is ignored if column is set. */
private boolean prefixMode=true;
/** How to select a read's key; see static modes array */
private int mode=AFFIX_MODE;
// /** Assume Illumina header format, and used the barcode. */
// private boolean mode==BARCODE_MODE=false;
//
// /** Allow names to be any substring of a header */
// private boolean substringMode=false;
//
// /** Demultiplex every sequence into its own file */
// private boolean mode==HEADER_MODE=false;
//
// /** Demultiplex mapped reads into one file per reference sequence */
// private boolean mode==CHROM_MODE=false;
/** Track per-output-file cardinality; may be slow */
private boolean trackCardinality=false;
/** Prevents issuing warnings multiple times */
private boolean warned=false;
/** Mcros buffers will be dumped after accumulating this many reads */
private int readsPerBuffer=1600;
/** Mcros buffers will be dumped after accumulating this many reads */
private int bytesPerBuffer=3000000;
/** Hamming distance for read indexes */
private int hdist=0;
/**
* Do not create files with under this many reads.
* Increases memory usage since reads must be retained until processing is finished.
* Upon running out of memory the program may go very slowly.
*/
private long minReadsToDump=0;
/** Affix length if fixed */
private int fixedLength=-1;
/** All possible affix lengths if there are multiple.
* Basically, the lengths of everything in names. */
private int[] lengthArray;
/**
* A set of recognized names and the output names they map to.
* The keys and values will be identical unless a Hamming distance is used.
* This is only filled if explicit names are provided.
*/
private HashMap<String, String> names=new HashMap<String, String>();
/** Names in list form, for substring matching */
private ArrayList<String> nameList=new ArrayList<String>();
/**
* Select MultiCros2 or MultiCros3.
* MultiCros2 only opens 1 stream at a time and does not do concurrent writes.
* MultiCros3 maintains multiple streams and does concurrent writes even with only 1 stream.
*/
private int mcrosType=3;
/** Max concurrent streams for MultiCros3. Does not seem to speed things up. */
private int maxStreams=BufferedMultiCros.DEFAULT_MAX_STREAMS;
/**
* This puts the mcros handling in a second thread.
* Does not seem to speed things up.
*/
private boolean threaded=false;
/** Whether input file interleaving was explicitly set */
private boolean setInterleaved=false;
/*--------------------------------------------------------------*/
/** Print messages here */
private PrintStream outstream=System.err;
/** True if errors were encountered */
public boolean errorState=false;
/** Permission to overwrite existing files. */
private boolean overwrite=true;
/**
* Append to existing files rather than overwriting.
* It is not advisable to set this flag for this class.
* */
private boolean append=false;
/** Retain header of sam/bam files. */
private boolean useSharedHeader=false;
/*--------------------------------------------------------------*/
/*---------------- Static Fields ----------------*/
/*--------------------------------------------------------------*/
/** Operation mode for selection of key */
public static final int AFFIX_MODE=1, DELIMITER_MODE=2, BARCODE_MODE=3, SUBSTRING_MODE=4, HEADER_MODE=5, CHROM_MODE=6;
/**
* Symbols allowed to substitute for Hamming distance.
* Hamming distance is intended for barcodes only.
*/
private static final byte[] symbols={'A', 'C', 'G', 'T', 'N'};
/** Verbose messages for debugging */
public static boolean verbose=false;
}
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