File: Realign.java

package info (click to toggle)
bbmap 39.20%2Bdfsg-3
  • links: PTS, VCS
  • area: main
  • in suites: forky, sid
  • size: 26,024 kB
  • sloc: java: 312,743; sh: 18,099; python: 5,247; ansic: 2,074; perl: 96; makefile: 39; xml: 38
file content (578 lines) | stat: -rwxr-xr-x 18,678 bytes parent folder | download | duplicates (2)
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124
125
126
127
128
129
130
131
132
133
134
135
136
137
138
139
140
141
142
143
144
145
146
147
148
149
150
151
152
153
154
155
156
157
158
159
160
161
162
163
164
165
166
167
168
169
170
171
172
173
174
175
176
177
178
179
180
181
182
183
184
185
186
187
188
189
190
191
192
193
194
195
196
197
198
199
200
201
202
203
204
205
206
207
208
209
210
211
212
213
214
215
216
217
218
219
220
221
222
223
224
225
226
227
228
229
230
231
232
233
234
235
236
237
238
239
240
241
242
243
244
245
246
247
248
249
250
251
252
253
254
255
256
257
258
259
260
261
262
263
264
265
266
267
268
269
270
271
272
273
274
275
276
277
278
279
280
281
282
283
284
285
286
287
288
289
290
291
292
293
294
295
296
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
341
342
343
344
345
346
347
348
349
350
351
352
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
372
373
374
375
376
377
378
379
380
381
382
383
384
385
386
387
388
389
390
391
392
393
394
395
396
397
398
399
400
401
402
403
404
405
406
407
408
409
410
411
412
413
414
415
416
417
418
419
420
421
422
423
424
425
426
427
428
429
430
431
432
433
434
435
436
437
438
439
440
441
442
443
444
445
446
447
448
449
450
451
452
453
454
455
456
457
458
459
460
461
462
463
464
465
466
467
468
469
470
471
472
473
474
475
476
477
478
479
480
481
482
483
484
485
486
487
488
489
490
491
492
493
494
495
496
497
498
499
500
501
502
503
504
505
506
507
508
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
533
534
535
536
537
538
539
540
541
542
543
544
545
546
547
548
549
550
551
552
553
554
555
556
557
558
559
560
561
562
563
564
565
566
567
568
569
570
571
572
573
574
575
576
577
578
package var2;

import java.io.PrintStream;
import java.util.ArrayList;

import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import shared.TrimRead;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FastaReadInputStream;
import stream.Read;
import stream.ReadStreamWriter;
import stream.SamLine;
import structures.ListNum;
import tracker.ReadStats;

/**
 * Realigns samlines to a reference.
 * 
 * @author Brian Bushnell
 * @date April 26, 2017
 *
 */
public class Realign {
	
	/*--------------------------------------------------------------*/
	/*----------------        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
		Realign x=new Realign(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 Realign(String[] args){
		
		{//Preparse block for help, config files, and outstream
			PreParser pp=new PreParser(args, getClass(), false);
			args=pp.args;
			outstream=pp.outstream;
		}
		
		//Set static variables
		SamLine.PARSE_OPTIONAL_MD_ONLY=true; //I only need the MD tag..
		SamLine.RNAME_AS_BYTES=false;
		SamLine.SET_FROM_OK=true;
		ReadStreamWriter.USE_ATTACHED_SAMLINE=true;
		ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
		ReadWrite.setZipThreads(Shared.threads());
		Shared.TRIM_READ_COMMENTS=Shared.TRIM_RNAME=true;
		
		//Create a parser object
		Parser parser=new Parser();
		parser.qtrimLeft=qtrimLeft;
		parser.qtrimRight=qtrimRight;
		parser.trimq=trimq;

		samFilter.includeUnmapped=false;
		samFilter.includeSupplimentary=false;
		samFilter.includeDuplicate=true;
		samFilter.includeNonPrimary=true;
		samFilter.includeQfail=false;
		samFilter.minMapq=4;
		
		//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.equals("parse_flag_goes_here")){
				long fake_variable=Parse.parseKMG(b);
				//Set a variable here
			}
			
			
			else if(a.equals("unclip")){
				unclip=Parse.parseBoolean(b);
			}else if(a.equals("realignrows") || a.equals("rerows")){
				Realigner.defaultMaxrows=Integer.parseInt(b);
			}else if(a.equals("realigncols") || a.equals("recols")){
				Realigner.defaultColumns=Integer.parseInt(b);
			}else if(a.equals("realignpadding") || a.equals("repadding") || a.equals("padding")){
				Realigner.defaultPadding=Integer.parseInt(b);
			}else if(a.equals("msa")){
				Realigner.defaultMsaType=b;
			}else if(a.equals("ref")){
				ref=b;
			}else if(a.equals("border")){
				border=Integer.parseInt(b);
			}
			
			else if(samFilter.parse(arg, a, b)){
				//do nothing
			}
			
			
			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];
			}
		}
		
		samFilter.setSamtoolsFilter();
		
		{//Process parser fields
			Parser.processQuality();
			
			maxReads=parser.maxReads;
			
			overwrite=ReadStats.overwrite=parser.overwrite;
			append=ReadStats.append=parser.append;
			
			in1=parser.in1;

			out1=parser.out1;
			
			trimq=parser.trimq;
			trimE=parser.trimE();
			
			extin=parser.extin;
			extout=parser.extout;
		}
		
		assert(FastaReadInputStream.settingsOK());
		
		//Ensure there is an input file
		if(in1==null || ref==null){throw new RuntimeException("Error - one input file and one reference are 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 output files can be written
		if(!Tools.testOutputFiles(overwrite, append, false, out1)){
			outstream.println((out1==null)+", "+out1);
			throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+out1+"\n");
		}
		
		//Ensure input files can be read
		if(!Tools.testInputFiles(false, true, in1, ref)){
			throw new RuntimeException("\nCan't read some input files.\n");  
		}
		
		//Ensure that no file was specified multiple times
		if(!Tools.testForDuplicateFiles(true, in1, ref, out1)){
			throw new RuntimeException("\nSome file names were specified multiple times.\n");
		}
		
		//Create output FileFormat objects
		ffout1=FileFormat.testOutput(out1, FileFormat.SAM, extout, true, overwrite, append, ordered);

		//Create input FileFormat objects
		ffin1=FileFormat.testInput(in1, FileFormat.SAM, extin, true, true);
		
		loadReference();
	}

	private void loadReference(){
		if(loadedRef){return;}
		assert(ref!=null);
		ScafMap.loadReference(ref, scafMap, samFilter, true);
		Realigner.map=scafMap;
		loadedRef=true;
	}
	
	/*--------------------------------------------------------------*/
	/*----------------         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, null, null, null);
			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 a read output stream
		final ConcurrentReadOutputStream ros;
		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() && (in1!=null && !ffin1.samOrBam() && !ffout1.samOrBam())){
				outstream.println("Writing interleaved.");
			}
			
			ros=ConcurrentReadOutputStream.getStream(ffout1, null, null, null, buff, null, true);
			ros.start(); //Start the stream
		}else{ros=null;}
		
		//Reset counters
		readsProcessed=0;
		basesProcessed=0;
		
		//Process the reads in separate threads
		spawnThreads(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);
		
		//Reset read validation
		Read.VALIDATE_IN_CONSTRUCTOR=vic;
		
		//Report timing and results
		t.stop();
		outstream.println("Realignments:  \t"+realignmentsAttempted);
		outstream.println("Successes:     \t"+realignmentsSucceeded);
		outstream.println("Improvements:  \t"+realignmentsImproved);
		outstream.println("Retained:      \t"+realignmentsRetained);
		outstream.println("Bases trimmed: \t"+basesTrimmed);
		outstream.println();
		outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
		
		//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 ros){
		
		//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, ros, 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;
			
			trimmedBasesProcessed+=pt.trimmedBasesProcessedT;
			basesTrimmed+=pt.basesTrimmedT;
			readsDiscarded+=pt.readsDiscardedT;
			pairedInSequencingReadsProcessed+=pt.pairedInSequencingReadsProcessedT;
			properlyPairedReadsProcessed+=pt.properlyPairedReadsProcessedT;

			realignmentsAttempted+=pt.realigner.realignmentsAttempted;
			realignmentsImproved+=pt.realigner.realignmentsImproved;
			realignmentsSucceeded+=pt.realigner.realignmentsSucceeded;
			realignmentsRetained+=pt.realigner.realignmentsRetained;
			
			success&=pt.success;
		}
		
		//Track whether any threads failed
		if(!success){errorState=true;}
		
		//Do anything necessary after processing
		
	}
	
	/*--------------------------------------------------------------*/
	/*----------------         Inner Methods        ----------------*/
	/*--------------------------------------------------------------*/
	
	/*--------------------------------------------------------------*/
	/*----------------         Inner Classes        ----------------*/
	/*--------------------------------------------------------------*/
	
	/** This class is static to prevent accidental writing to shared variables.
	 * It is safe to remove the static modifier. */
	private class ProcessThread extends Thread {
		
		//Constructor
		ProcessThread(final ConcurrentReadInputStream cris_, final ConcurrentReadOutputStream ros_, final int tid_){
			cris=cris_;
			ros=ros_;
			tid=tid_;
			realigner=new Realigner();
		}
		
		//Called by start()
		@Override
		public void run(){
			//Do anything necessary prior to processing
			
			//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);

			//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

				//Loop through each read in the list
				for(int idx=0; idx<reads.size(); idx++){
					final Read r1=reads.get(idx);
					
					//Validate reads in worker threads
					if(!r1.validated()){r1.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.
						boolean keep=processRead(r1);
						if(!keep){
							reads.set(idx, null);
							readsDiscardedT++;
						}
					}
				}

				//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

				//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());
			}
		}
		
		/**
		 * Process a read.
		 * @param r Read 1
		 * @return True if the reads should be kept, false if they should be discarded.
		 */
		boolean processRead(final Read r){
			if(r.bases==null || r.length()<=1){return false;}
			final SamLine sl=r.samline;
			final int len0=r.length();
			
			if(samFilter!=null && !samFilter.passesFilter(sl)){return false;}
			
			if(sl.properPair()){properlyPairedReadsProcessedT++;}
			if(sl.hasMate()){pairedInSequencingReadsProcessedT++;}
			final Scaffold scaf=scafMap.getScaffold(sl);
			final int scafnum=scaf.number;
			
//			r.toLongMatchString(false); //Not necessary if scoring can be done on short match string
			assert(sl.cigar!=null) : sl;
			boolean realigned=realigner.realign(r, sl, scaf, unclip);
			if(!realigned && r.match!=null){sl.cigar=SamLine.toCigar14(r.match, r.start, r.stop, scaf.length, sl.seq);}
			assert(sl.cigar!=null) : sl;
			
			int leftTrimAmount=border, rightTrimAmount=border;
			if(qtrimLeft || qtrimRight){
				long packed=TrimRead.testOptimal(r.bases, r.quality, trimE);
				if(qtrimLeft){leftTrimAmount=Tools.max(leftTrimAmount, (int)((packed>>32)&0xFFFFFFFFL));}
				if(qtrimRight){rightTrimAmount=Tools.max(rightTrimAmount, (int)((packed)&0xFFFFFFFFL));}
			}
			if(len0-leftTrimAmount-rightTrimAmount<2){return false;}
			
			int trimmed=(leftTrimAmount<1 && rightTrimAmount<1 ? 0 : TrimRead.trimReadWithMatch(r, sl, leftTrimAmount, rightTrimAmount, 0, scaf.length, false));
			if(trimmed<0 || r.length()<2 || sl.cigar==null){return false;}
			basesTrimmedT+=trimmed;
			if(trimmed>0){sl.optional=null;}
			
			assert(sl.cigar!=null) : leftTrimAmount+", "+rightTrimAmount+", "+len0+"\n"+sl;
			return true;
		}
		
		/** Number of reads processed by this thread */
		protected long readsProcessedT=0;
		/** Number of bases processed by this thread */
		protected long basesProcessedT=0;
		/** Number of trimmed, mapped bases processed. */
		protected long trimmedBasesProcessedT=0;
		/** Number of trimmed, mapped bases processed. */
		protected long basesTrimmedT=0;
		/** Number of reads discarded by this thread */
		protected long readsDiscardedT=0;
		/** Number of paired reads processed by this thread, whether or not they mapped as pairs */
		protected long pairedInSequencingReadsProcessedT=0;
		/** Number of properly paired reads processed by this thread */
		protected long properlyPairedReadsProcessedT=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 ros;
		/** For realigning reads */
		final Realigner realigner;
		
		/** Thread ID */
		final int tid;
	}
	
	/*--------------------------------------------------------------*/
	/*----------------            Fields            ----------------*/
	/*--------------------------------------------------------------*/

	/** Primary input file path */
	private String in1=null;

	/** A fasta file. */
	private String ref=null;
	
	/** Primary output file path */
	private String out1=null;
	
	/** Override input file extension */
	private String extin=null;
	/** Override output file extension */
	private String extout=null;
	
	/*--------------------------------------------------------------*/

	/** Number of reads processed */
	protected long readsProcessed=0;
	/** Number of bases processed */
	protected long basesProcessed=0;

	/** Quit after processing this many input reads; -1 means no limit */
	private long maxReads=-1;
	
	/*--------------------------------------------------------------*/
	
	
	private boolean loadedRef=false;

	private boolean qtrimLeft=false;
	private boolean qtrimRight=true;
	private float trimq=10;
	private final float trimE;
	
	/*--------------------------------------------------------------*/
	
	/** Number of trimmed, mapped bases processed */
	protected long trimmedBasesProcessed=0;
	/** Number of reads discarded */
	protected long readsDiscarded=0;
	/** Number of paired reads processed by this thread, whether or not they mapped as pairs */
	protected long pairedInSequencingReadsProcessed=0;
	/** Number of properly paired reads processed */
	protected long properlyPairedReadsProcessed=0;
	/** Number of bases trimmed */
	protected long basesTrimmed=0;
	
	protected long realignmentsAttempted;
	protected long realignmentsImproved;
	protected long realignmentsSucceeded;
	protected long realignmentsRetained;
	
	public final ScafMap scafMap=new ScafMap();
	
	public int border=0;
	public boolean unclip=false;
	
	public final SamFilter samFilter=new SamFilter();
	
	/*--------------------------------------------------------------*/
	/*----------------         Final Fields         ----------------*/
	/*--------------------------------------------------------------*/

	/** Primary input file */
	private final FileFormat ffin1;
	
	/** Primary output file */
	private final FileFormat ffout1;
	
	/*--------------------------------------------------------------*/
	/*----------------        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=true;
	
}