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package jgi;
import java.io.PrintStream;
import java.util.ArrayList;
import fileIO.ByteFile;
import fileIO.ByteStreamWriter;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.KillSwitch;
import shared.MetadataWriter;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import sort.ReadLengthComparator;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import structures.ListNum;
import tracker.ReadStats;
/**
* Reformats a fungal assembly for release. Also creates contig and agp files.
*
* @author Brian Bushnell
* @date December 9, 2015
*
*/
public class FungalRelease {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
*
* @param args
* Command line arguments
*/
public static void main(String[] args) {
Timer t = new Timer();
FungalRelease x = new FungalRelease(args);
x.process(t);
// Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
/**
* Constructor.
*
* @param args
* Command line arguments
*/
public FungalRelease(String[] args) {
{// Preparse block for help, config files, and outstream
PreParser pp = new PreParser(args, getClass(), false);
args = pp.args;
outstream = pp.outstream;
}
FASTQ.FORCE_INTERLEAVED = FASTQ.TEST_INTERLEAVED = false;
Shared.FASTA_WRAP = 60;
// Set shared static variables
Shared.capBuffers(4);
ReadWrite.USE_PIGZ = ReadWrite.USE_UNPIGZ = true;
ReadWrite.setZipThreads(Shared.threads());
Read.TO_UPPER_CASE = true;
// Create a parser object
Parser parser = new Parser();
// 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 (parser.parse(arg, a, b)) {// Parse standard flags in the parser
// do nothing
} else if (a.equals("verbose")) {
verbose = Parse.parseBoolean(b);
} else if (a.equals("mingapin")) {
minGapIn = (int) Parse.parseKMG(b);
} else if (a.equals("mingap") || a.equals("mingapout")) {
minGapOut = (int) Parse.parseKMG(b);
} else if (a.equals("minlen") || a.equals("minlength") || a.equals("minscaf")) {
minScaf = (int) Parse.parseKMG(b);
} else if (a.equals("mincontig")) {
minContig = (int) Parse.parseKMG(b);
} else if (a.equals("outc") || a.equals("contigs")) {
outC = b;
} else if (a.equals("qfoutc")) {
qfoutC = b;
} else if (a.equals("sortcontigs")) {
sortContigs = Parse.parseBoolean(b);
} else if (a.equals("sortcscaffolds")) {
sortScaffolds = Parse.parseBoolean(b);
} else if (a.equals("baniupac")) {
banIupac = Parse.parseBoolean(b);
} else if (a.equals("agp")) {
agpFile = b;
} else if (a.equals("legend")) {
legendFile = b;
} else if (a.equals("scafnum")) {
scafNum = Parse.parseKMG(b);
} else if (a.equals("renamescaffolds") || a.equals("rename")) {
renameScaffolds = Parse.parseBoolean(b);
} else if (a.equals("scafnum")) {
contigNum = Parse.parseKMG(b);
} else if (a.equals("renamecontigs")) {
renameContigs = Parse.parseBoolean(b);
} else if (a.equals("parse_flag_goes_here")) {
// Set a variable here
} else {
outstream.println("Unknown parameter " + args[i]);
assert (false) : "Unknown parameter " + args[i];
// throw new RuntimeException("Unknown parameter "+args[i]);
}
}
{// Process parser fields
Parser.processQuality();
maxReads = parser.maxReads;
overwrite = ReadStats.overwrite = parser.overwrite;
append = ReadStats.append = parser.append;
in1 = parser.in1;
qfin1 = parser.qfin1;
out1 = parser.out1;
qfout1 = parser.qfout1;
extin = parser.extin;
extout = parser.extout;
}
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 output files can be written
if (!Tools.testOutputFiles(overwrite, append, false, out1, outC)) {
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)) {
throw new RuntimeException("\nCan't read some input files.\n");
}
// Ensure that no file was specified multiple times
if (!Tools.testForDuplicateFiles(true, in1, out1, outC)) {
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
// Create output FileFormat objects
ffout1 = FileFormat.testOutput(out1, FileFormat.FASTA, extout, true, overwrite, append, false);
// Create output FileFormat objects
ffoutC = FileFormat.testOutput(outC, FileFormat.FASTA, extout, true, overwrite, append, false);
// Create input FileFormat objects
ffin1 = FileFormat.testInput(in1, FileFormat.FASTA, extin, true, true);
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
void process(Timer t) {
// Create a read input stream
final ConcurrentReadInputStream cris;
{
cris = ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, null, qfin1, null);
cris.start(); // Start the stream
if (verbose) {
outstream.println("Started cris");
}
}
// Optionally create a read output stream
final ConcurrentReadOutputStream ros, rosc;
final int buff = 4;
if (ffout1 != null) {
ros = ConcurrentReadOutputStream.getStream(ffout1, null, qfout1, null, buff, null, false);
ros.start(); // Start the stream
} else {
ros = null;
}
if (ffoutC != null) {
rosc = ConcurrentReadOutputStream.getStream(ffoutC, null, qfoutC, null, 4, null, false);
rosc.start(); // Start the stream
} else {
rosc = null;
}
// Reset counters
readsProcessed = 0;
basesProcessed = 0;
readsOut = 0;
basesOut = 0;
// Process the read stream
processInner(cris, ros, 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, ros, rosc);
// Report timing and results
t.stop();
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
MetadataWriter.write(null, readsProcessed, basesProcessed, readsOut, basesOut, 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.");
}
}
/** Iterate through the reads */
void processInner(final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream ros,
final ConcurrentReadOutputStream rosc) {
ArrayList<Read> scaffolds = getReads(cris);
final boolean makeLegend = (legendFile != null);
ByteStreamWriter bswLegend = (makeLegend ? new ByteStreamWriter(legendFile, overwrite, append, false) : null);
if (bswLegend != null) {
bswLegend.start();
}
if (ros != null) {
if (sortScaffolds) {
Shared.sort(scaffolds, ReadLengthComparator.comparator);
}
if (renameScaffolds) {
for (Read r : scaffolds) {
String old = r.id;
r.id = "scaffold_" + scafNum;
if (bswLegend != null) {
bswLegend.println(old + "\t" + r.id);
}
scafNum++;
}
}
ros.add(scaffolds, 0);
}
if (bswLegend != null) {
bswLegend.poisonAndWait();
}
final boolean makeAgp = (agpFile != null);
ByteStreamWriter bswAgp = (makeAgp ? new ByteStreamWriter(agpFile, overwrite, append, false) : null);
if (bswAgp != null) {
bswAgp.start();
}
if (rosc != null || makeAgp) {// Process contigs
ArrayList<Read> contigs = new ArrayList<Read>();
for (Read r : scaffolds) {
ArrayList<Read> temp = r.breakAtGaps(makeAgp, minContig);
if (bswAgp != null) {
bswAgp.print((byte[]) r.obj);
r.obj=null;
}
contigs.addAll(temp);
}
if (sortContigs) {
Shared.sort(contigs, ReadLengthComparator.comparator);
}
if (renameContigs) {
for (Read r : contigs) {
r.id = "contig_" + contigNum;
contigNum++;
}
}
if (rosc != null) {
rosc.add(contigs, 0);
}
}
if (bswAgp != null) {
bswAgp.poisonAndWait();
}
}
/** Iterate through the reads */
private ArrayList<Read> getReads(final ConcurrentReadInputStream cris) {
ArrayList<Read> all = new ArrayList<Read>(10000);
{
// 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 == null || ffin1.samOrBam()) || (r.mate != null) == cris.paired());
}
// 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.");
}
// Loop through each read in the list
for (int idx = 0; idx < reads.size(); idx++) {
final Read r1 = reads.get(idx);
assert (r1.mate == null);
// Track the initial length for statistics
final int initialLength1 = r1.length();
// Increment counters
readsProcessed+=1;
basesProcessed+=initialLength1;
boolean keep = processRead(r1);
if (keep) {
all.add(r1);
readsOut+=1;
basesOut+=initialLength1;
}
}
// Notify the input stream that the list was used
cris.returnList(ln);
if (verbose) {
outstream.println("Returned a list.");
}
// 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());
}
}
return all;
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
/**
* Process a single read pair.
*
* @param r1
* Read 1
* @return True if the reads should be kept, false if they should be
* discarded.
*/
boolean processRead(final Read r1) {
//assert (!banIupac || !r1.containsNonACGTN()) : "Non-ACGTN base found in scaffold " + r1.id;
if(banIupac){
if(r1.containsNonACGTN()){
KillSwitch.exceptionKill(new RuntimeException("Non-ACGTN base found in scaffold " + r1.id));
}
}
r1.inflateGaps(minGapIn, minGapOut);
return r1.length() >= minScaf;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
private int minGapIn = 1;
private int minGapOut = 10;
private int minScaf = 1;
private int minContig = 1;
private long scafNum = 1;
private long contigNum = 1;
private boolean sortScaffolds = true;
private boolean sortContigs = false;
private boolean banIupac = true;
private boolean renameScaffolds = true;
private boolean renameContigs = false;
/*--------------------------------------------------------------*/
/*---------------- I/O Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file path */
private String in1 = null;
private String qfin1 = null;
/** Primary output file path */
private String out1 = null;
private String outC = null;
private String qfout1 = null;
private String qfoutC = null;
private String agpFile = null;
private String legendFile = 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;
/** Number of reads processed */
protected long readsOut = 0;
/** Number of bases processed */
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;
/** Primary output file */
private final FileFormat ffout1;
private final FileFormat ffoutC;
/*--------------------------------------------------------------*/
/*---------------- 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 = false;
/** 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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