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package jgi;
import java.util.ArrayList;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import fileIO.TextStreamWriter;
import kmer.AbstractKmerTable;
import kmer.TableLoaderLockFree;
import kmer.TableReader;
import shared.Parse;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.Read;
import structures.ListNum;
import template.BBTool_ST;
/**
* @author Brian Bushnell
* @date Mar 2, 2015
*
*/
public class SplitNexteraLMP extends BBTool_ST {
public static void main(String[] args){
Timer t=new Timer();
SplitNexteraLMP bbt=new SplitNexteraLMP(args);
bbt.process(t);
}
@Override
protected void setDefaults(){
outStats="stderr";
minReadLength=40;
mask=false;
merge=false;
testmerge=0;
pairedInput=true;
symbol='J';
useInnerLMP=false;
RENAME=true;
}
/**
* @param args
*/
public SplitNexteraLMP(String[] args) {
super(args);
reparse(args);
tables=(mask ? TableLoaderLockFree.makeTables(AbstractKmerTable.ARRAY1D, 12, -1L, false, 0.9) : null);
if(outFrag1!=null && outFrag2==null && outFrag1.indexOf('#')>-1){
outFrag2=outFrag1.replace("#", "2");
outFrag1=outFrag1.replace("#", "1");
}
if(outUnk1!=null && outUnk2==null && outUnk1.indexOf('#')>-1){
outUnk2=outUnk1.replace("#", "2");
outUnk1=outUnk1.replace("#", "1");
}
if(testmerge>0){
System.err.println("Testing merge rate.");
float rate=FileFormat.testInput(in1, FileFormat.FASTQ, null, true, true).stdio() ? 1 : BBMerge.mergeableFraction(in1, in2, 1000000, 0.2f);
merge=rate>0.1;
System.err.println("Merge rate: "+Tools.format("%.2f%%", rate));
if(!merge){
System.err.println("Merging was disabled due to a low merge rate of "+Tools.format("%.3f", rate));
}
}
}
@Override
public boolean parseArgument(String arg, String a, String b) {
if(a.equals("symbol") || a.equals("junction")){
assert(b!=null && b.length()==1) : "Junction symbol must be a single character.";
symbol=(byte)b.charAt(0);
return true;
}else if(a.equals("outfrag") || a.equals("outfrag1") || a.equals("outf") || a.equals("outf1")){
outFrag1=b;
return true;
}else if(a.equals("outfrag2") || a.equals("outf2")){
outFrag2=b;
return true;
}else if(a.equals("outunknown") || a.equals("outunknown1") || a.equals("outu") || a.equals("outu1")){
outUnk1=b;
return true;
}else if(a.equals("outunknown2") || a.equals("outu2")){
outUnk2=b;
return true;
}else if(a.equals("outsingle") || a.equals("outs")){
outSingle=b;
return true;
}else if(a.equals("minlen") || a.equals("minlength") || a.equals("ml")){
minReadLength=Integer.parseInt(b);
return true;
}else if(a.equals("useinnerlmp") || a.equals("innerlmp")){
useInnerLMP=Parse.parseBoolean(b);
return true;
}else if(a.equals("rename")){
RENAME=Parse.parseBoolean(b);
return true;
}else if(a.equals("literal")){
literals=(b==null ? null : b.split(","));
return true;
}else if(a.equals("mask")){
mask=Parse.parseBoolean(b);
return true;
}else if(a.equals("merge")){
merge=Parse.parseBoolean(b);
return true;
}else if(a.equals("testmerge")){
testmerge=Double.parseDouble(b);
if(testmerge>1){testmerge/=100;}
return true;
}else if(a.equals("rcomp")){
rcomp=Parse.parseBoolean(b);
return true;
}else if(a.equals("maskmiddle") || a.equals("mm")){
maskMiddle=Parse.parseBoolean(b);
return true;
}else if(a.equals("k")){
k=Integer.parseInt(b);
return true;
}else if(a.equals("mink")){
mink=Integer.parseInt(b);
return true;
}else if(a.equals("hdist") || a.equals("hammingdistance")){
hdist=Integer.parseInt(b);
return true;
}else if(a.equals("hdist2") || a.equals("hammingdistance2")){
hdist2=Integer.parseInt(b);
return true;
}else if(a.equals("edits") || a.equals("edist") || a.equals("editdistance")){
edist=Integer.parseInt(b);
return true;
}else if(a.equals("dump")){
dump=b;
return true;
}else if(a.equals("stats")){
outStats=b;
return true;
}
return false;
}
@Override
protected void startupSubclass(){
if(!Tools.testOutputFiles(overwrite, append, false, out1, out2, outFrag1, outFrag2, outUnk1, outUnk2)){
throw new RuntimeException("\noverwrite="+overwrite+", append="+append+"\n" +
"Can't write to output files "+out1+", "+out2+", "+outFrag1+", "+outFrag2+", "+outUnk1+", "+outUnk2+"\n");
}
if(!Tools.testForDuplicateFiles(true, in1, in2, qfin1, qfin2, out1, out2, qfout1, qfout2, outFrag1, outFrag2, outUnk1, outUnk2)){
assert(false) : "Duplicate files.";
}
ffoutFrag1=FileFormat.testOutput(outFrag1, FileFormat.FASTQ, extout, true, overwrite, append, false);
ffoutFrag2=FileFormat.testOutput(outFrag2, FileFormat.FASTQ, extout, true, overwrite, append, false);
ffoutUnk1=FileFormat.testOutput(outUnk1, FileFormat.FASTQ, extout, true, overwrite, append, false);
ffoutUnk2=FileFormat.testOutput(outUnk2, FileFormat.FASTQ, extout, true, overwrite, append, false);
ffoutSingle=FileFormat.testOutput(outSingle, FileFormat.FASTQ, extout, true, overwrite, append, false);
if(outFrag1!=null){
final int buff=4;
rosFrag=ConcurrentReadOutputStream.getStream(ffoutFrag1, ffoutFrag2, null, null, buff, null, false);
rosFrag.start();
}else{rosFrag=null;}
if(outUnk1!=null){
final int buff=4;
rosUnk=ConcurrentReadOutputStream.getStream(ffoutUnk1, ffoutUnk2, null, null, buff, null, false);
rosUnk.start();
}else{rosUnk=null;}
if(outSingle!=null){
final int buff=4;
rosSingle=ConcurrentReadOutputStream.getStream(ffoutSingle, null, null, null, buff, null, false);
rosSingle.start();
}else{rosSingle=null;}
}
@Override
/** Iterate through the reads.
* This may optionally be overridden. */
protected void processInner(final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream rosLmp){
if(mask){
final TableLoaderLockFree loader=new TableLoaderLockFree(tables, k, mink, 0, hdist, edist, rcomp, maskMiddle);
loader.setRefSkip(0);
loader.hammingDistance2=hdist2;
loader.storeMode(TableLoaderLockFree.SET_IF_NOT_PRESENT);
long kmers=loader.processData(null, literals, false, false, false);
outstream.println("Added "+kmers+" kmers.");
if(dump!=null){
ByteStreamWriter bsw=new ByteStreamWriter(dump, overwrite, false, true);
bsw.start();
for(AbstractKmerTable set : tables){
set.dumpKmersAsBytes(bsw, k, 0, Integer.MAX_VALUE, null);
}
bsw.poisonAndWait();
}
reader=new TableReader(k, mink, 0, 0, 0, rcomp, maskMiddle);
reader.trimSymbol=symbol;
assert(kmers>0) : "There were no stored kmers; please check your settings.";
// assert(false) : hdist+", "+hdist2+", "+maskMiddle+", "+rcomp+", "+k+", "+mink;
}else{
reader=null;
}
readsProcessed=0;
basesProcessed=0;
readsLmp=0;
basesLmp=0;
readsFrag=0;
basesFrag=0;
readsUnk=0;
basesUnk=0;
pairedInput=cris.paired();
{
ListNum<Read> ln=cris.nextList();
ArrayList<Read> listIn=(ln!=null ? ln.list : null);
if(listIn!=null && !listIn.isEmpty()){
Read r=listIn.get(0);
assert((ffin1==null || ffin1.samOrBam()) || (r.mate!=null)==cris.paired());
}
while(listIn!=null && listIn.size()>0){
if(verbose){outstream.println("Fetched "+listIn.size()+" reads.");}
ArrayList<Read> outLmp=new ArrayList<Read>(listIn.size());
ArrayList<Read> outFrag=new ArrayList<Read>(listIn.size());
ArrayList<Read> outUnk=new ArrayList<Read>(listIn.size());
ArrayList<Read> outSingle=new ArrayList<Read>(listIn.size());
for(int idx=0; idx<listIn.size(); idx++){
final Read r1=listIn.get(idx);
final Read r2=r1.mate;
final int initialLength1=r1.length();
final int initialLength2=(r1.mateLength());
readsProcessed+=r1.pairCount();
basesProcessed+=initialLength1+initialLength2;
boolean keep=processReadPair(r1, r2, outLmp, outFrag, outUnk, outSingle);
}
for(Read r1 : outLmp){
readsLmp+=r1.pairCount();
basesLmp+=r1.pairLength();
assert(r1.mate!=null);
assert(r1.pairnum()==0);
assert(r1.mate.pairnum()==1) : r1.mate.id+"\n\n"+r1.length()+"\n"+r1+"\n\n"+r1.mateLength()+"\n"+r1.mate+"\n\n";
assert(r1.mate.mate==r1);
}
if(rosLmp!=null){rosLmp.add(outLmp, ln.id);}
for(Read r1 : outFrag){
readsFrag+=r1.pairCount();
basesFrag+=r1.pairLength();
assert(r1.mate!=null);
assert(r1.pairnum()==0);
assert(r1.mate.pairnum()==1);
assert(r1.mate.mate==r1);
}
if(rosFrag!=null){rosFrag.add(outFrag, ln.id);}
for(Read r1 : outUnk){
readsUnk+=r1.pairCount();
basesUnk+=r1.pairLength();
assert(r1.mate!=null);
assert(r1.pairnum()==0);
assert(r1.mate.pairnum()==1);
assert(r1.mate.mate==r1);
}
if(rosUnk!=null){rosUnk.add(outUnk, ln.id);}
for(Read r1 : outSingle){
readsSingle+=r1.pairCount();
basesSingle+=r1.pairLength();
assert(r1.pairnum()==0);
assert(r1.mate==null);
}
if(rosSingle!=null){rosSingle.add(outSingle, ln.id);}
cris.returnList(ln);
if(verbose){outstream.println("Returned a list.");}
ln=cris.nextList();
listIn=(ln!=null ? ln.list : null);
}
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
}
@Override
protected void showStatsSubclass(final Timer t, long readsIn, long basesIn){
TextStreamWriter tsw=new TextStreamWriter(outStats==null ? "stderr" : outStats, overwrite, append, false);
tsw.start();
outstream.println("");
final double rmult=(pairedInput ? 100.0 : 50.0)/readsIn;
final double bmult=100.0/basesIn;
//Note that this can go over 100%
tsw.println("Long Mate Pairs: \t"+readsLmp+" reads ("+Tools.format("%.2f",readsLmp*rmult)+"%) \t"+
basesLmp+" bases ("+Tools.format("%.2f",basesLmp*bmult)+"%)");
tsw.println("Fragment Pairs: \t"+readsFrag+" reads ("+Tools.format("%.2f",readsFrag*rmult)+"%) \t"+
basesFrag+" bases ("+Tools.format("%.2f",basesFrag*bmult)+"%)");
tsw.println("Unknown Pairs: \t"+readsUnk+" reads ("+Tools.format("%.2f",readsUnk*rmult)+"%) \t"+
basesUnk+" bases ("+Tools.format("%.2f",basesUnk*bmult)+"%)");
tsw.println("Singletons: \t"+readsSingle+" reads ("+Tools.format("%.2f",readsSingle*100.0/readsIn)+"%) \t"+
basesSingle+" bases ("+Tools.format("%.2f",basesSingle*bmult)+"%)");
tsw.println("\n(Note: Read totals may exceed 100%, though base totals should not.)");
tsw.println("");
tsw.println("Adapters Detected: \t"+junctionsDetected+" ("+Tools.format("%.2f%%)",junctionsDetected*100.0/junctionsSought));
tsw.println("Bases Recovered: \t"+(basesLmp+basesFrag+basesUnk+basesSingle)+
" ("+Tools.format("%.2f%%)",(basesLmp+basesFrag+basesUnk+basesSingle)*bmult));
if(merge){
tsw.println("");
tsw.println("Merged Pairs: \t"+mergedReadCount+" ("+Tools.format("%.2f%%)",mergedReadCount*200.0/readsProcessed));
tsw.println("Merged Bases: \t"+mergedBaseCount+" ("+Tools.format("%.2f%%)",mergedBaseCount*100.0/basesProcessed));
}
errorState|=tsw.poisonAndWait();
}
@Override
protected boolean processReadPair(Read r1, Read r2) {
throw new RuntimeException("Do not use.");
}
boolean processReadPair(Read r1, Read r2, ArrayList<Read> outLmp, ArrayList<Read> outFrag, ArrayList<Read> outUnk, ArrayList<Read> outSingle) {
boolean needsMasking=mask;
if(merge){
int insert=BBMerge.findOverlapStrict(r1, r2, false);
if(insert>0){
r2.reverseComplement();
Read merged=r1.joinRead(insert);
r2.reverseComplement();
int a=1, b=0, c=0;
if(mask){
a=reader.kMask(merged, tables);
}
if(a>0 || true){
mergedReadCount++;
mergedBaseCount+=r1.length()+r2.length()-merged.length();
return processMergedRead(merged, r1, r2, outLmp, outFrag, outUnk, outSingle);
}else if(mask){
needsMasking=false;
b=reader.kMask(r1, tables);
c=reader.kMask(r2, tables);
if(b==0 && c==0){
mergedReadCount++;
mergedBaseCount+=r1.length()+r2.length()-merged.length();
return processMergedRead(merged, r1, r2, outLmp, outFrag, outUnk, outSingle);
}
}
}
}
if(needsMasking){
int a=reader.kMask(r1, tables);
int b=reader.kMask(r2, tables);
}
junctionsSought++;
r1.start=Tools.indexOf(r1.bases, symbol);
r1.stop=Tools.lastIndexOf(r1.bases, symbol);
assert(r1==null || r1.pairnum()==0);
assert(r2==null || r2.pairnum()==1);
if(r2!=null){
r2.start=Tools.indexOf(r2.bases, symbol);
r2.stop=Tools.lastIndexOf(r2.bases, symbol);
if(r1.start<0 && r2.start<0){
if(verbose){System.err.println("Added unknown pair "+r1.id);}
outUnk.add(r1);
return true;
}
r1.mate=r2.mate=null;
}else if(r1.start<0){
if(verbose){System.err.println("Added singleton "+r1.id);}
outSingle.add(r1);
return true;
}
junctionsDetected++;
Read r1left=null, r1right=null, r2left=null, r2right=null;
// final Read r1left, r1right, r2left, r2right;
if(r2==null){
if(r1.start>=0){
int left=r1.start;
int right=r1.length()-r1.stop-1;
r1left=(left>=minReadLength ? r1.subRead(0, r1.start) : null);
r1right=null;
r2left=null;
r2right=(right>=minReadLength ? r1.subRead(r1.stop+1, r1.length()) : null);
if(r2right!=null){
r2right.setPairnum(1);
if(RENAME){
r2right.id=r2right.id.replaceFirst(" /1", " /2");
r2right.id=r2right.id.replaceFirst(" 1:", " 2:");
}
}
}
}else if(r1.start>=0 && r2.start>=0){//confusing
{
int left=r1.start;
int right=r1.length()-r1.stop-1;
r1left=(left>=minReadLength ? r1.subRead(0, r1.start) : null);
r1right=(right>=minReadLength ? r1.subRead(r1.stop+1, r1.length()) : null);
}
{
int left=r2.start;
int right=r2.length()-r2.stop-1;
//Note these are reversed
r2left=(right>=minReadLength ? r2.subRead(r2.stop+1, r2.length()) : null);
r2right=(left>=minReadLength ? r2.subRead(0, r2.start) : null);
}
}else if(r1.start>=0){
int left=r1.start;
int right=r1.length()-r1.stop-1;
r1left=(left>=minReadLength ? r1.subRead(0, r1.start) : null);
r1right=(right>=minReadLength ? r1.subRead(r1.stop+1, r1.length()) : null);
r2left=null;
r2right=r2;
}else if(r2.start>=0){
int left=r2.start;
int right=r2.length()-r2.stop-1;
//Note these are reversed
r2left=(right>=minReadLength ? r2.subRead(r2.stop+1, r2.length()) : null);
r2right=(left>=minReadLength ? r2.subRead(0, r2.start) : null);
r1left=r1;
r1right=null;
}else{
assert(false) : r1.start+", "+r1.stop+(r2==null ? "null" : ", "+r2.start+", "+r2.stop);
}
boolean outerLMP=false, innerLMP=false, leftFrag=false, rightFrag=false;
if(r1left!=null && r2right!=null){//outer lmp
if(verbose){System.err.println("Added outer LMP "+r1.id);}
r1left.mate=r2right;
r2right.mate=r1left;
outLmp.add(r1left);
r1left=r2right=null;
outerLMP=true;
}
if(r1right!=null && r2left!=null){//inner lmp
if(verbose){System.err.println("Added inner LMP "+r1.id);}
if(useInnerLMP){
r1right.mate=r2left;
r2left.mate=r1right;
outLmp.add(r1right);
r1right=r2left=null;
innerLMP=true;
}
}
if(r1left!=null && r2left!=null){//left frag
if(verbose){System.err.println("Added left frag "+r1.id);}
r1left.mate=r2left;
r2left.mate=r1left;
outFrag.add(r1left);
r1left=r2left=null;
leftFrag=true;
}
if(r1right!=null && r2right!=null){//right frag
if(verbose){System.err.println("Added right frag "+r1.id);}
r1right.mate=r2right;
r2right.mate=r1right;
outFrag.add(r1right);
r1right=r2right=null;
rightFrag=true;
}
//Singletons
if(r1left!=null){
if(verbose){System.err.println("Added singleton r1left "+r1left.id);}
outSingle.add(r1left);
}
if(r1right!=null){
if(verbose){System.err.println("Added singleton r1right "+r1right.id);}
outSingle.add(r1right);
}
if(r2left!=null){
if(verbose){System.err.println("Added singleton r2left "+r2left.id);}
r2left.setPairnum(0);
outSingle.add(r2left);
}
if(r2right!=null){
if(verbose){System.err.println("Added singleton r2right "+r2right.id);}
r2right.setPairnum(0);
outSingle.add(r2right);
}
return true;
}
boolean processMergedRead(Read merged, Read r1, Read r2, ArrayList<Read> outLmp, ArrayList<Read> outFrag, ArrayList<Read> outUnk, ArrayList<Read> outSingle) {
// int a=0, b, c;
// if(mask){
// a=reader.kMask(merged, tables);
// }
junctionsSought++;
merged.start=Tools.indexOf(merged.bases, symbol);
merged.stop=Tools.lastIndexOf(merged.bases, symbol);
assert(merged!=null && merged.pairnum()==0);
if(merged.start<0){
if(verbose){System.err.println("Added frag "+r1.id);}
outFrag.add(r1);
return true;
}
junctionsDetected++;
Read r1left=null, r1right=null;
int left=merged.start;
int right=merged.length()-merged.stop-1;
r1left=(left>=minReadLength ? merged.subRead(0, merged.start) : null);
r1right=(right>=minReadLength ? merged.subRead(merged.stop+1, merged.length()-1) : null);
if(r1right!=null && r1left!=null){
r1right.setPairnum(1);
if(RENAME){
r1right.id=r1right.id.replaceFirst(" /1", " /2");
r1right.id=r1right.id.replaceFirst(" 1:", " 2:");
}
if(verbose){System.err.println("Added outer LMP "+merged.id);}
r1left.mate=r1right;
r1right.mate=r1left;
outLmp.add(r1left);
return true;
}
//Singletons
if(r1left!=null){
if(verbose){System.err.println("Added singleton r1left "+r1left.id);}
outSingle.add(r1left);
}
if(r1right!=null){
if(verbose){System.err.println("Added singleton r1right "+r1right.id);}
outSingle.add(r1right);
}
return true;
}
@Override
protected void shutdownSubclass(){
errorState|=ReadWrite.closeStreams(null, rosFrag, rosUnk, rosSingle);
}
@Override
protected final boolean useSharedHeader(){return true;}
/*--------------------------------------------------------------*/
/*---------------- Masking Fields ----------------*/
/*--------------------------------------------------------------*/
private String[] literals=new String[] {"CTGTCTCTTATACACATCTAGATGTGTATAAGAGACAG"};
private final AbstractKmerTable[] tables;
private int k=19;
private int mink=11;
private int hdist=1;
private int hdist2=0;
private int edist=0;
private boolean rcomp=true;
private boolean maskMiddle=false;
private String dump=null;
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
private TableReader reader;
protected String outStats="stderr";
protected String outFrag1;
protected String outFrag2;
protected String outUnk1;
protected String outUnk2;
protected String outSingle;
protected FileFormat ffoutFrag1;
protected FileFormat ffoutFrag2;
protected FileFormat ffoutUnk1;
protected FileFormat ffoutUnk2;
protected FileFormat ffoutSingle;
protected ConcurrentReadOutputStream rosFrag;
protected ConcurrentReadOutputStream rosUnk;
protected ConcurrentReadOutputStream rosSingle;
private int minReadLength;
private boolean mask;
private boolean merge;
private double testmerge;
public long readsLmp(){return readsLmp;}
public long basesLmp(){return basesLmp;}
public long readsFrag(){return readsFrag;}
public long basesFrag(){return basesFrag;}
public long readsUnk(){return readsUnk;}
public long basesUnk(){return basesUnk;}
public long readsSingle(){return readsSingle;}
public long basesSingle(){return basesSingle;}
private long readsLmp=0;
private long basesLmp=0;
private long readsFrag=0;
private long basesFrag=0;
private long readsUnk=0;
private long basesUnk=0;
private long readsSingle=0;
private long basesSingle=0;
private long mergedReadCount=0;
private long mergedBaseCount=0;
private long junctionsSought=0, junctionsDetected=0;
private boolean pairedInput;
private byte symbol;
private boolean useInnerLMP;
private boolean RENAME;
}
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