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package jgi;
import java.io.File;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.Arrays;
import dna.AminoAcid;
import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.KillSwitch;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import structures.ByteBuilder;
import structures.ListNum;
import tracker.ReadStats;
/**
* This class does nothing.
* It is designed to be easily modified into a program
* that processes reads in a single thread.
*
* @author Brian Bushnell
* @date June 20, 2014
*
*/
public class AdjustHomopolymers {
/*--------------------------------------------------------------*/
/*---------------- 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
AdjustHomopolymers x=new AdjustHomopolymers(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 AdjustHomopolymers(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
//Set shared static variables prior to parsing
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Shared.threads());
Shared.capBuffers(4); //Only for singlethreaded programs
{//Parse the arguments
final Parser parser=parse(args);
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;
extin=parser.extin;
out1=parser.out1;
out2=parser.out2;
qfout1=parser.qfout1;
qfout2=parser.qfout2;
extout=parser.extout;
}
assert(rate!=0) : "'rate' should be set to a positive or negative value, such as rate=0.1 to expand homopolymers by 10%.";
doPoundReplacement(); //Replace # with 1 and 2
adjustInterleaving(); //Make sure interleaving agrees with number of input and output files
fixExtensions(); //Add or remove .gz or .bz2 as needed
checkFileExistence(); //Ensure files can be read and written
checkStatics(); //Adjust file-related static fields as needed for this program
//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 input FileFormat objects
ffin1=FileFormat.testInput(in1, FileFormat.FASTQ, extin, true, true);
ffin2=FileFormat.testInput(in2, FileFormat.FASTQ, extin, true, true);
}
/*--------------------------------------------------------------*/
/*---------------- Initialization Helpers ----------------*/
/*--------------------------------------------------------------*/
/** Parse arguments from the command line */
private Parser parse(String[] args){
//Create a parser object
Parser parser=new Parser();
//Set any necessary Parser defaults here
//parser.foo=bar;
//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(b!=null && b.equalsIgnoreCase("null")){b=null;}
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
}else if(a.equals("rate")){
rate=Float.parseFloat(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];
}
}
return parser;
}
/** Replace # with 1 and 2 in headers */
private void doPoundReplacement(){
//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");
}
//Ensure there is an input file
if(in1==null){throw new RuntimeException("Error - at least one input file is required.");}
//Ensure out2 is not set without out1
if(out1==null && out2!=null){throw new RuntimeException("Error - cannot define out2 without defining out1.");}
}
/** Add or remove .gz or .bz2 as needed */
private void fixExtensions(){
in1=Tools.fixExtension(in1);
in2=Tools.fixExtension(in2);
qfin1=Tools.fixExtension(qfin1);
qfin2=Tools.fixExtension(qfin2);
}
/** Ensure files can be read and written */
private void checkFileExistence(){
//Ensure output files can be written
if(!Tools.testOutputFiles(overwrite, append, false, out1, out2)){
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)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
}
/** Make sure interleaving agrees with number of input and output files */
private void adjustInterleaving(){
//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;
}
//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);
}
}
}
}
/** Adjust file-related static fields as needed for this program */
private static void checkStatics(){
//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;
}
assert(FastaReadInputStream.settingsOK());
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
void process(Timer t){
//Create a read input stream
final ConcurrentReadInputStream cris=makeCris();
//Optionally create a read output stream
final ConcurrentReadOutputStream ros=makeCros(cris.paired());
//Reset counters
readsProcessed=readsOut=0;
basesProcessed=basesOut=0;
//Process the read stream
processInner(cris, ros);
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, ros);
//Report timing and results
t.stop();
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
outstream.println(Tools.readsBasesOut(readsProcessed, basesProcessed, readsOut, basesOut, 8, false));
//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.");
}
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
private ConcurrentReadInputStream makeCris(){
ConcurrentReadInputStream 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"));}
return cris;
}
private ConcurrentReadOutputStream makeCros(boolean pairedInput){
if(ffout1==null){return null;}
//Select output buffer size based on whether it needs to be ordered
final int buff=4;
//Notify user of output mode
if(pairedInput && out2==null && (in1!=null && !ffin1.samOrBam() && !ffout1.samOrBam())){
outstream.println("Writing interleaved.");
}
final ConcurrentReadOutputStream ros=ConcurrentReadOutputStream.getStream(ffout1, ffout2, qfout1, qfout2, buff, null, false);
ros.start(); //Start the stream
return ros;
}
/** Iterate through the reads */
void processInner(final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream ros){
//Do anything necessary prior to processing
{
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Check to ensure pairing is as expected
if(ln!=null && !ln.isEmpty()){
Read r=ln.get(0);
assert(ffin1.samOrBam() || (r.mate!=null)==cris.paired());
}
//As long as there is a nonempty read list...
while(ln!=null && ln.size()>0){
// if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
processList(ln, cris, ros);
//Fetch a new list
ln=cris.nextList();
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
//Do anything necessary after processing
}
/**
* Process a list of Reads.
* @param ln The list.
* @param cris Read Input Stream
* @param ros Read Output Stream for reads that will be retained
*/
void processList(ListNum<Read> ln, final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream ros){
//Grab the actual read list from the ListNum
final ArrayList<Read> reads=ln.list;
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r1=reads.get(idx);
final Read r2=r1.mate;
//Validate reads in worker threads
if(!r1.validated()){r1.validate(true);}
if(r2!=null && !r2.validated()){r2.validate(true);}
//Track the initial length for statistics
final int initialLength1=r1.length();
final int initialLength2=r1.mateLength();
//Increment counters
readsProcessed+=r1.pairCount();
basesProcessed+=initialLength1+initialLength2;
{
//Reads are processed in this block.
boolean keep=processReadPair(r1, r2);
if(!keep){reads.set(idx, null);}
else{
readsOut+=r1.pairCount();
basesOut+=r1.pairLength();
}
}
}
//Output reads to the output stream
if(ros!=null){ros.add(reads, 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
}
/**
* Process a single read pair.
* @param r1 Read 1
* @param r2 Read 2 (may be null)
* @return True if the reads should be kept, false if they should be discarded.
*/
boolean processReadPair(final Read r1, final Read r2){
processRead(r1);
processRead(r2);
return true;
}
void processRead(final Read r){
if(r==null){return;}
bbBases.clear();
bbQuals.clear();
final byte[] bases=r.bases;
final byte[] quals=(r.quality==null ? fakeQuality(bases.length) : r.quality);
byte prevBase='?';
byte prevQual=20;
byte streak=0;
for(int i=0; i<bases.length; i++){
final byte b=bases[i];
final byte q=quals[i];
bbBases.append(b);
bbQuals.append(q);
if(b==prevBase){
streak++;
}else{
int adjustment=AminoAcid.isFullyDefined(prevBase) ? (int)(rate*streak) : 0;
if(adjustment<0){
bbBases.length+=adjustment;
bbQuals.length+=adjustment;
}else{
for(; adjustment>0; adjustment--){
bbBases.append(prevBase);
bbQuals.append(prevQual);
}
}
streak=1;
}
prevBase=b;
prevQual=q;
}
if(bbBases.length()!=r.length()){
r.bases=bbBases.toBytes();
r.quality=(r.quality==null ? null : bbQuals.toBytes());
}
}
private byte[] fakeQuality(int minlen){
if(fakeQuality.length<minlen){
fakeQuality=KillSwitch.allocByte1D(minlen+10);
Arrays.fill(fakeQuality, Shared.FAKE_QUAL);
}
return fakeQuality;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** 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;
private String qfout1=null;
private String qfout2=null;
/** Override input file extension */
private String extin=null;
/** Override output file extension */
private String extout=null;
/** Whether interleaved was explicitly set. */
private boolean setInterleaved=false;
private ByteBuilder bbBases=new ByteBuilder();
private ByteBuilder bbQuals=new ByteBuilder();
private float rate=0.0f;
private byte[] fakeQuality=new byte[0];
/*--------------------------------------------------------------*/
/** 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 */
private final FileFormat ffin1;
/** Secondary input file */
private final FileFormat ffin2;
/** Primary output file */
private final FileFormat ffout1;
/** Secondary output file */
private final FileFormat ffout2;
/*--------------------------------------------------------------*/
/*---------------- 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;
/** This flag has no effect on singlethreaded programs */
private final boolean ordered=false;
}
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