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*
* created: 2009
*
* This file is part of Artemis
*
* Copyright(C) 2009 Genome Research Limited
*
* This program is free software; you can redistribute it and/or
* modify it under the terms of the GNU General Public License
* as published by the Free Software Foundation; either version 2
* of the License, or(at your option) any later version.
*
* This program is distributed in the hope that it will be useful,
* but WITHOUT ANY WARRANTY; without even the implied warranty of
* MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the
* GNU General Public License for more details.
*
* You should have received a copy of the GNU General Public License
* along with this program; if not, write to the Free Software
* Foundation, Inc., 59 Temple Place - Suite 330, Boston, MA 02111-1307, USA.
*
*/
import java.awt.AlphaComposite;
import java.awt.BasicStroke;
import java.awt.BorderLayout;
import java.awt.Color;
import java.awt.Component;
import java.awt.Composite;
import java.awt.GridBagConstraints;
import java.awt.GridBagLayout;
import java.awt.event.ActionEvent;
import java.awt.event.ActionListener;
import java.awt.event.AdjustmentEvent;
import java.awt.event.AdjustmentListener;
import java.awt.event.ItemEvent;
import java.awt.event.ItemListener;
import java.awt.event.KeyAdapter;
import java.awt.event.KeyEvent;
import java.awt.image.BufferedImage;
import java.io.FileOutputStream;
import java.io.IOException;
import java.lang.management.ManagementFactory;
import java.lang.management.MemoryMXBean;
import java.util.ArrayList;
import java.util.concurrent.CountDownLatch;
import java.util.concurrent.ExecutorService;
import java.util.concurrent.Executors;
import javax.swing.ButtonGroup;
import javax.swing.JCheckBoxMenuItem;
import javax.swing.JComponent;
import javax.swing.JLabel;
import javax.swing.JMenu;
import javax.swing.JMenuItem;
import javax.swing.JRadioButton;
import javax.swing.JSeparator;
import javax.swing.border.Border;
import javax.swing.border.EmptyBorder;
import javax.swing.event.ChangeEvent;
import javax.swing.event.ChangeListener;
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import org.apache.log4j.Level;
import net.sf.picard.reference.ReferenceSequenceFile;
import net.sf.picard.sam.BuildBamIndex;
import net.sf.samtools.AlignmentBlock;
import net.sf.samtools.SAMException;
import net.sf.samtools.SAMFileHeader;
import net.sf.samtools.SAMFileReader;
import net.sf.samtools.SAMRecord;
import net.sf.samtools.SAMSequenceRecord;
import net.sf.samtools.SAMFileReader.ValidationStringency;
import net.sf.samtools.util.CloseableIterator;
import uk.ac.sanger.artemis.Entry;
import uk.ac.sanger.artemis.EntryGroup;
import uk.ac.sanger.artemis.FeatureVector;
import uk.ac.sanger.artemis.Selection;
import uk.ac.sanger.artemis.SelectionChangeEvent;
import uk.ac.sanger.artemis.SelectionChangeListener;
import uk.ac.sanger.artemis.circular.TextFieldInt;
import uk.ac.sanger.artemis.components.DisplayAdjustmentEvent;
import uk.ac.sanger.artemis.components.DisplayAdjustmentListener;
import uk.ac.sanger.artemis.components.EntryEdit;
import uk.ac.sanger.artemis.components.FeatureDisplay;
tjc
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import uk.ac.sanger.artemis.components.FileViewer;
import uk.ac.sanger.artemis.components.IndexReferenceEvent;
import uk.ac.sanger.artemis.components.NonModalDialog;
import uk.ac.sanger.artemis.components.SequenceComboBox;
import uk.ac.sanger.artemis.components.SwingWorker;
import uk.ac.sanger.artemis.editor.MultiLineToolTipUI;
import uk.ac.sanger.artemis.sequence.MarkerRange;
import uk.ac.sanger.artemis.sequence.NoSequenceException;
import uk.ac.sanger.artemis.util.Document;
import uk.ac.sanger.artemis.util.DocumentFactory;
import uk.ac.sanger.artemis.util.FTPSeekableStream;
public class BamView extends JPanel
implements DisplayAdjustmentListener, SelectionChangeListener
private List<BamViewRecord> readsInView;
private Hashtable<String, SAMFileReader> samFileReaderHash = new Hashtable<String, SAMFileReader>();
private List<SAMReadGroupRecord> readGroups = new Vector<SAMReadGroupRecord>();
private HashMap<String, Integer> seqLengths = new HashMap<String, Integer>();
private HashMap<String, Integer> offsetLengths;
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protected List<String> bamList;
private boolean isSingle = false;
private boolean isSNPs = false;
private FeatureDisplay feature_display;
private Selection selection;
private boolean logScale = false;
private Ruler ruler;
private int startBase = -1;
private int endBase = -1;
private boolean showBaseAlignment = false;
private JMenu bamFilesMenu = new JMenu("BAM files");
private JCheckBoxMenuItem logMenuItem = new JCheckBoxMenuItem("Use Log Scale", logScale);
private JCheckBoxMenuItem cbStackView = new JCheckBoxMenuItem("Stack", true);
private JCheckBoxMenuItem cbPairedStackView = new JCheckBoxMenuItem("Paired Stack");
private JCheckBoxMenuItem cbStrandStackView = new JCheckBoxMenuItem("Strand Stack");
private JCheckBoxMenuItem cbIsizeStackView = new JCheckBoxMenuItem("Inferred Size", false);
private JCheckBoxMenuItem cbCoverageView = new JCheckBoxMenuItem("Coverage", false);
private JCheckBoxMenuItem cbCoverageStrandView = new JCheckBoxMenuItem("Coverage by Strand", false);
private JCheckBoxMenuItem cbCoverageHeatMap = new JCheckBoxMenuItem("Coverage Heat Map", false);
private JCheckBoxMenuItem cbLastSelected;
private ButtonGroup buttonGroup = new ButtonGroup();
private JCheckBoxMenuItem colourByReadGrp = new JCheckBoxMenuItem("Read Group");
private JCheckBoxMenuItem colourByStrandTag = new JCheckBoxMenuItem("RNASeq Strand Specific Tag (XS)");
private JCheckBoxMenuItem colourByCoverageColour = new JCheckBoxMenuItem("Coverage Plot Colours");
private JCheckBoxMenuItem baseQualityColour = new JCheckBoxMenuItem("Base Quality");
private JCheckBoxMenuItem markInsertions = new JCheckBoxMenuItem("Mark Insertions", true);
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private AlphaComposite translucent =
AlphaComposite.getInstance(AlphaComposite.SRC_OVER, 0.6f);
private GroupBamFrame groupsFrame = new GroupBamFrame(this, bamFilesMenu);
private CoveragePanel coverageView = new CoveragePanel();
protected static String BAM_SUFFIX = ".*\\.(bam|cram)$";
private static Color LIGHT_GREY = new Color(200, 200, 200);
private static Color DARK_GREEN = new Color(0, 150, 0);
private static Color DARK_ORANGE = new Color(255,140,0);
private static Color DEEP_PINK = new Color(139,10,80);
private static Color NON_SELECTED_READ_HIGHLIGHT_COLOUR = new Color(189,103,107);
private volatile BamViewRecord mouseOverSAMRecord = null;
private volatile BamViewRecord highlightSAMRecord = null;
private String mouseOverInsertion;
// record of where a mouse drag starts
private static int MAX_BASES = 26000;
private int maxHeight = 800;
tjc
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private int BASE_HEIGHT;
private PopupMessageFrame popFrame = new PopupMessageFrame();
private PopupMessageFrame waitingFrame = new PopupMessageFrame("waiting...");
private ExecutorService bamReadTaskExecutor;
private int MAX_COVERAGE = Integer.MAX_VALUE;
public static org.apache.log4j.Logger logger4j =
org.apache.log4j.Logger.getLogger(BamView.class);
public BamView(List<String> bamList,
String reference,
int nbasesInView,
final EntryEdit entry_edit,
final FeatureDisplay feature_display,
final Bases bases,
final JPanel containerPanel,
final JFrame frame)
{
this(bamList, reference, nbasesInView, feature_display, bases, containerPanel, frame);
this.entry_edit = entry_edit;
}
public BamView(List<String> bamList,
int nbasesInView,
final FeatureDisplay feature_display,
final Bases bases,
this.bamList = bamList;
this.feature_display = feature_display;
this.bases = bases;
containerPanel.setLayout(new BoxLayout(containerPanel, BoxLayout.Y_AXIS));
containerPanel.add(mainPanel);
// filter out unmapped reads by default
setSamRecordFlagPredicate(
new SAMRecordFlagPredicate(SAMRecordFlagPredicate.READ_UNMAPPED_FLAG));
System.setProperty("reference", reference); // for CRAM
try
{
getEntry(reference,entryGroup);
}
catch (NoSequenceException e)
{
e.printStackTrace();
}
}
if(Options.getOptions().getIntegerProperty("bam_read_thread") != null)
{
logger4j.debug("BAM READ THREADS="+Options.getOptions().getIntegerProperty("bam_read_thread"));
bamReadTaskExecutor = Executors.newFixedThreadPool(
Options.getOptions().getIntegerProperty("bam_read_thread"));
}
else
bamReadTaskExecutor = Executors.newFixedThreadPool(1);
if(Options.getOptions().getIntegerProperty("bam_max_coverage") != null)
{
logger4j.debug("BAM MAX COVERAGE="+Options.getOptions().getIntegerProperty("bam_max_coverage"));
MAX_COVERAGE = Options.getOptions().getIntegerProperty("bam_max_coverage");
}
try
{
readHeaderPicard();
}
catch(java.lang.UnsupportedClassVersionError err)
{
JOptionPane.showMessageDialog(null,
"This requires Java 1.6 or higher.",
"Check Java Version", JOptionPane.WARNING_MESSAGE);
}
catch (IOException e)
{
e.printStackTrace();
}
final javax.swing.plaf.FontUIResource font_ui_resource =
Options.getOptions().getFontUIResource();
while(keys.hasMoreElements())
{
Object key = keys.nextElement();
Object value = UIManager.get(key);
if(value instanceof javax.swing.plaf.FontUIResource)
UIManager.put(key, font_ui_resource);
}
setFont(Options.getOptions().getFont());
ALIGNMENT_PIX_PER_BASE = fm.charWidth('M');
BASE_HEIGHT = fm.getMaxAscent();
MultiLineToolTipUI.initialize();
setToolTipText("");
buttonGroup.add(cbStackView);
buttonGroup.add(cbPairedStackView);
buttonGroup.add(cbStrandStackView);
buttonGroup.add(cbIsizeStackView);
buttonGroup.add(cbCoverageView);
buttonGroup.add(cbCoverageStrandView);
jspView = new JScrollPane(this,
JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED,
JScrollPane.HORIZONTAL_SCROLLBAR_NEVER);
jspView.setViewportBorder(BorderFactory.createMatteBorder(0, 0, 1, 0, Color.DARK_GRAY));
Border empty = new EmptyBorder(0,0,0,0);
jspView.setBorder(empty);
jspView.getVerticalScrollBar().setUnitIncrement(8);
// apply command line options
if(System.getProperty("show_snps") != null)
isSNPs = true;
if(System.getProperty("show_snp_plot") != null)
{
isSNPplot = true;
snpPanel.setVisible(isSNPplot);
}
if(System.getProperty("show_cov_plot") != null)
{
isCoverage = true;
coveragePanel.setVisible(isCoverage);
}
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{
if(isCoverageView(getPixPerBaseByWidth()) && lastMousePoint != null)
return coverageView.getToolTipText(
lastMousePoint.y-getJspView().getViewport().getViewPosition().y);
if(mouseOverSAMRecord == null)
return null;
String msg =
mouseOverSAMRecord.sam.getReadName() + "\n" +
mouseOverSAMRecord.sam.getAlignmentStart() + ".." +
mouseOverSAMRecord.sam.getAlignmentEnd() +
(mouseOverSAMRecord.sam.getReadGroup() != null ? "\nRG="+mouseOverSAMRecord.sam.getReadGroup().getId() : "") +
"\nisize=" + mouseOverSAMRecord.sam.getInferredInsertSize() +
"\nmapq=" + mouseOverSAMRecord.sam.getMappingQuality()+
"\nrname="+ mouseOverSAMRecord.sam.getReferenceName();
if( mouseOverSAMRecord.sam.getReadPairedFlag() &&
mouseOverSAMRecord.sam.getProperPairFlag() &&
!mouseOverSAMRecord.sam.getMateUnmappedFlag())
{
msg = msg +
"\nstrand (read/mate): "+
(mouseOverSAMRecord.sam.getReadNegativeStrandFlag() ? "-" : "+")+" / "+
(mouseOverSAMRecord.sam.getMateNegativeStrandFlag() ? "-" : "+");
}
else
msg = msg +
"\nstrand (read/mate): "+
(mouseOverSAMRecord.sam.getReadNegativeStrandFlag() ? "-" : "+");
if(msg != null && mouseOverInsertion != null)
msg = msg + "\nInsertion at:" +mouseOverInsertion;
return msg;
/**
* Get the BAM index file from the list
* @param bam
* @return
* @throws IOException
*/
private File getBamIndexFile(String bam) throws IOException
{
File bamIndexFile = null;
if (bam.startsWith("http") || bam.startsWith("ftp"))
{
final URL urlBamIndexFile = new URL(bam + ".bai");
InputStream is = urlBamIndexFile.openStream();
// Create temp file.
bamIndexFile = File.createTempFile(urlBamIndexFile.getFile().replaceAll(
"[\\/\\s]", "_"), ".bai");
bamIndexFile.deleteOnExit();
FileOutputStream out = new FileOutputStream(bamIndexFile);
int c;
while ((c = is.read()) != -1)
out.write(c);
out.flush();
out.close();
is.close();
logger4j.debug("create... " + bamIndexFile.getAbsolutePath());
{
final File cramIndexFile = new File(bam + ".crai");
if(cramIndexFile.exists())
{
logger4j.debug(
"ERROR: CRAM INDEX FILE ("+cramIndexFile.getName()+
") EXPECTING A BAM INDEX FILE (USE THIS OPTION --bam-style-index) ");
/**
* Get the SAM file reader.
* @param bam
* @return
* @throws IOException
*/
private SAMFileReader getSAMFileReader(final String bam) throws IOException
{
// parsing of the header happens during SAMFileReader construction,
// so need to set the default stringency
SAMFileReader.setDefaultValidationStringency(ValidationStringency.LENIENT);
if(samFileReaderHash.containsKey(bam))
return samFileReaderHash.get(bam);
if(!bamIndexFile.exists())
{
try
{
logger4j.warn("Index file not found so using picard to index the BAM.");
// Use Picard to index the file
// requires reads to be sorted by coordinate
new BuildBamIndex().instanceMain(
new String[]{ "I="+bam, "O="+bamIndexFile.getAbsolutePath(), "MAX_RECORDS_IN_RAM=50000", "VALIDATION_STRINGENCY=SILENT" });
}
catch(SAMException e)
{
String ls = System.getProperty("line.separator");
String msg =
"BAM index file is missing. The BAM file needs to be sorted and indexed"+ls+
"This can be done using samtools (http://samtools.sf.net/):"+ls+ls+
"samtools sort <in.bam> <out.prefix>"+ls+
"samtools index <sorted.bam>";
throw new SAMException(msg);
}
if(feature_display != null && bam.endsWith("cram"))
{
// set log level
net.sf.picard.util.Log.setGlobalLogLevel(
net.sf.picard.util.Log.LogLevel.ERROR);
final CRAMReferenceSequenceFile ref = new CRAMReferenceSequenceFile(
feature_display.getEntryGroup().getSequenceEntry(), this);
final Map<Object, ReferenceSequenceFile> referenceFactory =
new HashMap<Object, ReferenceSequenceFile>();
referenceFactory.put(bamIndexFile, ref);
try
{
Class<?> cls = getClass().getClassLoader().loadClass("net.sf.samtools.ReferenceDiscovery");
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Field f = cls.getDeclaredField("referenceFactory");
f.set(null, referenceFactory);
}
catch (ClassNotFoundException e)
{
System.err.println("Check cramtools.jar is in the CLASSPATH. "+e.getMessage());
}
catch (SecurityException e)
{
e.printStackTrace();
}
catch (NoSuchFieldException e)
{
e.printStackTrace();
}
catch (IllegalArgumentException e)
{
e.printStackTrace();
}
catch (IllegalAccessException e)
{
e.printStackTrace();
}
//net.sf.samtools.ReferenceDiscovery.referenceFactory.put(bamIndexFile, ref);
}
if(bam.startsWith("ftp"))
{
FTPSeekableStream fss = new FTPSeekableStream(new URL(bam));
samFileReader = new SAMFileReader(fss, bamIndexFile, false);
}
else if(!bam.startsWith("http"))
samFileReader = new SAMFileReader(bamFile, bamIndexFile);
}
else
{
final URL urlBamFile = new URL(bam);
samFileReader = new SAMFileReader(urlBamFile, bamIndexFile, false);
samFileReader.setValidationStringency(ValidationStringency.SILENT);
samFileReaderHash.put(bam, samFileReader);
readGroups.addAll(samFileReader.getFileHeader().getReadGroups());
final SAMFileReader inputSam = getSAMFileReader(bamList.get(0));
final SAMFileHeader header = inputSam.getFileHeader();
for(SAMSequenceRecord seq: header.getSequenceDictionary().getSequences())
seqLengths.put(seq.getSequenceName(),
seq.getSequenceLength());
seqNames.add(seq.getSequenceName());
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class BamReadTask implements Runnable
{
private int start;
private int end;
private short bamIndex;
private float pixPerBase;
private CountDownLatch latch;
BamReadTask(int start, int end, short bamIndex, float pixPerBase, CountDownLatch latch)
{
this.start = start;
this.end = end;
this.bamIndex = bamIndex;
this.pixPerBase = pixPerBase;
this.latch = latch;
}
public void run()
{
try
{
readFromBamPicard(start, end, bamIndex, pixPerBase) ;
}
catch (OutOfMemoryError ome)
{
throw ome;
}
catch(IOException me)
{
me.printStackTrace();
}
finally
{
latch.countDown();
}
}
}
private void readFromBamPicard(int start, int end, short bamIndex, float pixPerBase)
{
// Open the input file. Automatically detects whether input is SAM or BAM
// and delegates to a reader implementation for the appropriate format.
final SAMFileReader inputSam = getSAMFileReader(bam);
//final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile);
int sBeg = offset+1;
int sEnd = sBeg+sLen-1;
if( (sBeg >= start && sBeg <= end) ||
(sBeg <= start && sEnd >= start) )
{
int thisStart = start - offset;
if(thisStart < 1)
thisStart = 1;
int thisEnd = end - offset;
if(thisEnd > sLen)
thisEnd = sLen;
iterateOverBam(inputSam, seq, thisStart, thisEnd, bamIndex, pixPerBase, bam);
//System.out.println("READ "+seq+" "+thisStart+".."+thisEnd+" "+start+" --- "+offset);
}
}
}
else
{
String refName = (String) combo.getSelectedItem();
iterateOverBam(inputSam, refName, start, end, bamIndex, pixPerBase, bam);
}
/**
* Iterate over BAM file and load into the <code>List</code> of
* <code>SAMRecord</code>.
* @param inputSam
* @param refName
* @param start
* @param end
*/
private void iterateOverBam(final SAMFileReader inputSam,
final String refName, final int start, final int end,
final short bamIndex, final float pixPerBase,
final String bam)
{
final MemoryMXBean memory = ManagementFactory.getMemoryMXBean();
final int seqOffset = getSequenceOffset(refName);
final int offset = seqOffset- getBaseAtStartOfView();
final boolean isCoverageView = isCoverageView(pixPerBase);
int nbins = 800;
int binSize = ((end-start)/nbins)+1;
if(binSize < 1)
{
binSize = 1;
nbins = end-start+1;
}
int max = MAX_COVERAGE*binSize;
if(max < 1)
max = Integer.MAX_VALUE;
final int cov[] = new int[nbins];
for(int i=0; i<nbins; i++)
cov[i] = 0;
final CloseableIterator<SAMRecord> it = inputSam.queryOverlapping(refName, start, end);
try
{
cnt++;
SAMRecord samRecord = it.next();
if(readGrpFrame != null && !readGrpFrame.isReadGroupVisible(samRecord.getReadGroup()))
continue;
if( samRecordFlagPredicate == null ||
!samRecordFlagPredicate.testPredicate(samRecord))
if(samRecordMapQPredicate == null ||
samRecordMapQPredicate.testPredicate(samRecord))
{
int abeg = samRecord.getAlignmentStart();
int aend = samRecord.getAlignmentEnd();
boolean over = false;
for(int i=abeg; i<aend; i++)
{
int bin = ((i-start)/binSize)-1;
if(bin < 0)
else if(bin > nbins-1)
bin = nbins-1;
cov[bin]++;
if(cov[bin] > max)
{
over = true;
break;
}
}
if(over)
continue;
if(isCoverageView)
coverageView.addRecord(samRecord, offset, bam, colourByStrandTag.isSelected());
coveragePanel.addRecord(samRecord, offset, bam, colourByStrandTag.isSelected());
if(isSNPplot)
snpPanel.addRecord(samRecord, seqOffset);
if(!isCoverageView)
readsInView.add(new BamViewRecord(samRecord, bamIndex));
}
cnt = 0;
float heapFraction =
(float)((float)memory.getHeapMemoryUsage().getUsed()/
(float)memory.getHeapMemoryUsage().getMax());
logger4j.debug("Heap memory usage (used/max): "+heapFraction);
if(readsInView.size() > checkMemAfter*2 && !waitingFrame.isVisible())
waitingFrame.showWaiting("loading...", mainPanel);
if(heapFraction > 0.90)
{
popFrame.show(
"Using > 90 % of the maximum memory limit:"+
(memory.getHeapMemoryUsage().getMax()/1000000.f)+" Mb.\n"+
"Not all reads in this range have been read in. Zoom in or\n"+
"consider increasing the memory for this application.",
mainPanel,
15000);
break;
}
catch(Exception e)
{
System.err.println(e.getMessage());
}
finally
{
it.close();
}
return len;
}
else
return seqLengths.get((String) combo.getSelectedItem());
}
/**
* For BAM files with multiple references sequences, calculate
* the offset from the start of the concatenated sequence for
* a given reference.
* @param refName
* @return
*/
protected int getSequenceOffset(String refName)
{
if(offsetLengths == null)
offsetLengths = new HashMap<String, Integer>(combo.getItemCount());
int offset = 0;
for(int i=0; i<combo.getItemCount(); i++)
{
String thisSeqName = (String) combo.getItemAt(i);
offsetLengths.put(thisSeqName, offset);
offset += seqLengths.get(combo.getItemAt(i));
}
return offsetLengths.get(refName);
}
final FeatureVector features = feature_display.getEntryGroup().getAllFeatures();
final HashMap<String, Integer> lookup = new HashMap<String, Integer>();
for(int i=0; i<features.size(); i++)
lookup.put(features.elementAt(i).getIDString(), features.elementAt(i).getFirstBase());
offsetLengths = new HashMap<String, Integer>(seqNames.size());
for(int i=0; i<seqNames.size(); i++)
{
final Integer pos = lookup.get(seqNames.get(i));
if(pos != null)
offsetLengths.put(seqNames.get(i), pos-1);
/*final FeatureContigPredicate predicate = new FeatureContigPredicate(seqNames.get(i).trim());
for(int j=0; j<features.size(); j++)
{
if(predicate.testPredicate(features.elementAt(j)))
{
offsetLengths.put(seqNames.get(i), features.elementAt(j).getFirstBase()-1);
break;
}
if(offsetLengths.size() != seqNames.size())
System.err.println("Found: "+offsetLengths.size() +" of "+ seqNames.size());
SwingUtilities.invokeLater(new Runnable()
{
public void run()
{
JOptionPane.showMessageDialog(BamView.this,
"There is a problem matching the reference sequences\n"+
"to the names in the BAM file. This may mean the labels\n"+
"on the reference features do not match those in the in\n"+
"the BAM file.",
"Problem Found", JOptionPane.WARNING_MESSAGE);
}
});
int offset = 0;
for(int i=0; i<combo.getItemCount(); i++)
{
String thisSeqName = (String) combo.getItemAt(i);
offsetLengths.put(thisSeqName, offset);
offset += seqLengths.get(combo.getItemAt(i));
}
//return 0;
return offsetLengths.get(refName);
protected void paintComponent(Graphics g)
{
super.paintComponent(g);
Graphics2D g2 = (Graphics2D)g;
if(startBase > 0)
start = startBase;
else
if(endBase > 0)
end = endBase;
else
end = start + nbasesInView - 1;
if(end > seqLength)
end = seqLength;
if(feature_display != null && nbasesInView < feature_display.getMaxVisibleBases())
nbasesInView = feature_display.getMaxVisibleBases();
boolean changeToStackView = false;
MemoryMXBean memory = ManagementFactory.getMemoryMXBean();
coverageView.init(this, pixPerBase, start, end);
if(isCoverage)
coveragePanel.init(this, pixPerBase, start, end);
if(isSNPplot)
snpPanel.init(this, pixPerBase, start, end);
synchronized (this)
try
{
float heapFractionUsedBefore = (float) ((float) memory.getHeapMemoryUsage().getUsed() /
(float) memory.getHeapMemoryUsage().getMax());
if(readsInView == null)
readsInView = new Vector<BamViewRecord>();
else
readsInView.clear();
final CountDownLatch latch = new CountDownLatch(bamList.size()-hideBamList.size());
//long ms = System.currentTimeMillis();
for(short i=0; i<bamList.size(); i++)
{
if(!hideBamList.contains(i))
bamReadTaskExecutor.execute(
new BamReadTask(start, end, i, pixPerBase, latch));
try
{
latch.await();
}
catch (InterruptedException e) {} // TODO
//System.out.println("===== NO. THREADS="+
// ((java.util.concurrent.ThreadPoolExecutor)bamReadTaskExecutor).getPoolSize()+" TIME="+(System.currentTimeMillis()-ms));
float heapFractionUsedAfter = (float) ((float) memory.getHeapMemoryUsage().getUsed() /
(float) memory.getHeapMemoryUsage().getMax());
// System.out.println("Heap Max : "+memory.getHeapMemoryUsage().getMax());
// System.out.println("Heap Used : "+memory.getHeapMemoryUsage().getUsed());
// System.out.println("Heap memory used "+heapFractionUsedAfter);
if ((heapFractionUsedAfter - heapFractionUsedBefore) > 0.06
&& !isStackView() && heapFractionUsedAfter > 0.8)
changeToStackView = true;
}
if((!isStackView() && !isStrandStackView()) || isBaseAlignmentView(pixPerBase))
{
Collections.sort(readsInView, new SAMRecordComparator());
}
else if( (isStackView() || isStrandStackView()) &&
bamList.size() > 1)
{
// merge multiple BAM files
Collections.sort(readsInView, new SAMRecordPositionComparator(BamView.this));
}
catch (OutOfMemoryError ome)
JOptionPane.showMessageDialog(this, "Out of Memory");
readsInView.clear();
return;
catch(net.sf.samtools.util.RuntimeIOException re)
{
JOptionPane.showMessageDialog(this, re.getMessage());
}
// this needs to be synchronized when cloning BAM window
synchronized(this)
{
if(showBaseAlignment)
drawBaseAlignment(g2, seqLength, pixPerBase, start, end);
{
if(isCoverageView(pixPerBase))
drawCoverage(g2,start, end, pixPerBase);
else if(isStackView())
drawStackView(g2, seqLength, pixPerBase, start, end);
else if(isPairedStackView())
drawPairedStackView(g2, seqLength, pixPerBase, start, end);
else if(isStrandStackView())
drawStrandStackView(g2, seqLength, pixPerBase, start, end);
else
drawLineView(g2, seqLength, pixPerBase, start, end);
}
}
if(isCoverage)
coveragePanel.repaint();
if(isSNPplot)
snpPanel.repaint();
if(changeToStackView)
{
"Note :: Changed to the stack view to save memory.\n"+
"Currently this is using "+
(memory.getHeapMemoryUsage().getUsed()/1000000.f)+" Mb "+
"and the maximum\nmemory limit is "+
(memory.getHeapMemoryUsage().getMax()/1000000.f)+" Mb.",
protected void repaintBamView()
{
laststart = -1;
repaint();
}
return (float)mainPanel.getWidth() / (float)nbasesInView;
{
if(feature_display == null)
return seqLength+nbasesInView/3;
/**
* Draw the zoomed-in base view.
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawBaseAlignment(Graphics2D g2,
int seqLength,
float pixPerBase,
final int start,
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int ypos = 0;
end = start + ( mainPanel.getWidth() * ALIGNMENT_PIX_PER_BASE );
if(seqEnd > bases.getLength())
seqEnd = bases.getLength();
if(refSeqStart < 1)
refSeqStart = 1;
refSeq =
bases.getSubSequence(new Range(refSeqStart, seqEnd), Bases.FORWARD).toUpperCase();
ruler.refSeq = refSeq;
}
catch (OutOfRangeException e)
{
e.printStackTrace();
}
}
ruler.repaint();
drawSelectionRange(g2, ALIGNMENT_PIX_PER_BASE, start, end, Color.PINK);
g2.setStroke(new BasicStroke (2.f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND));
Rectangle r = jspView.getViewport().getViewRect();
int nreads = readsInView.size();
BamViewRecord thisRead = readsInView.get(i);
if (ypos < r.getMaxY() || ypos > r.getMinY())
drawSequence(g2, thisRead, ypos, refSeq, refSeqStart);
drawn[i] = true;
int thisEnd = thisRead.sam.getAlignmentEnd();
thisEnd = thisRead.sam.getAlignmentStart() + thisRead.sam.getReadLength();
BamViewRecord nextRead = readsInView.get(j);
int nextStart = nextRead.sam.getAlignmentStart();
if (nextStart > thisEnd + 1)
{
if (ypos < r.getMaxY() || ypos > r.getMinY())
drawSequence(g2, nextRead, ypos, refSeq, refSeqStart);
drawn[j] = true;
thisEnd = nextRead.sam.getAlignmentEnd();
thisEnd = nextStart + nextRead.sam.getReadLength();
}
else if (ypos > r.getMaxY() || ypos < r.getMinY())
break;
catch (ArrayIndexOutOfBoundsException ae)
{
System.err.println(readsInView.size()+" "+nreads);
ae.printStackTrace();
}
Dimension d = getPreferredSize();
d.setSize(getPreferredSize().getWidth(), ypos);
setPreferredSize(d);
/**
* Draw the query sequence
* @param g2
* @param read
* @param pixPerBase
* @param ypos
*/
private void drawSequence(final Graphics2D g2, final BamViewRecord bamViewRecord,
int ypos, String refSeq, int refSeqStart)
SAMRecord samRecord = bamViewRecord.sam;
if (!samRecord.getReadPairedFlag() || // read is not paired in sequencing
samRecord.getMateUnmappedFlag() ) // mate is unmapped ) // mate is unmapped
final Color col = g2.getColor();
int len = 0;
int refPos = 0;
SAMRecordSequenceString readSeq = new SAMRecordSequenceString(samRecord.getReadString());
final int offset = getSequenceOffset(samRecord.getReferenceName());
byte[] phredQuality = null;
if(baseQualityColour.isSelected())
phredQuality = samRecord.getBaseQualities();
Hashtable<Integer, String> insertions = null;
List<AlignmentBlock> blocks = samRecord.getAlignmentBlocks();
for(int i=0; i<blocks.size(); i++)
{
AlignmentBlock block = blocks.get(i);
int blockStart = block.getReadStart();
for(int j=0; j<block.getLength(); j++)
int readPos = blockStart-1+j;
xpos = block.getReferenceStart() - 1 + j + offset;
refPos = xpos - refSeqStart + 1;
if(phredQuality != null && phredQuality.length > 0)
setColourByBaseQuality(g2, phredQuality[readPos]);
if(isSNPs && refSeq != null && refPos > 0 && refPos < refSeq.length())
if(
(!readSeq.isSecondaryAlignment()) &&
(Character.toUpperCase(readSeq.charAt(readPos)) != refSeq.charAt(refPos))
)
g2.setColor(Color.red);
else
g2.setColor(col);
}
g2.drawString(readSeq.substring(readPos, readPos+1),
refPos*ALIGNMENT_PIX_PER_BASE, ypos);
if(isSNPs)
g2.setColor(col);
// look for insertions
if(markInsertions.isSelected() && i < blocks.size()-1)
{
int blockEnd = blockStart+block.getLength();
int nextBlockStart = blocks.get(i+1).getReadStart();
int insertSize = nextBlockStart - blockEnd;
if(insertSize > 0)
{
if(insertions == null)
insertions = new Hashtable<Integer, String>();
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(refPos+refSeqStart+1)+" "+
readSeq.substring(blockEnd-1, nextBlockStart-1));
g2.drawLine(xscreen, ypos, xscreen, ypos-BASE_HEIGHT);
// mark on reference sequence as well
if(bases != null)
g2.drawLine(xscreen, 11, xscreen, 11-BASE_HEIGHT);
g2.setColor(col);
}
}
if(highlightSAMRecord != null &&
highlightSAMRecord.sam.getReadName().equals(bamViewRecord.sam.getReadName()))
{
refPos = block.getReferenceStart() + offset - refSeqStart;
int xstart = refPos*ALIGNMENT_PIX_PER_BASE;
int width = block.getLength()*ALIGNMENT_PIX_PER_BASE;
Color col1 = g2.getColor();
//g2.setColor(Color.red);
if (isThisBamRecordHighlighted(bamViewRecord) )
{
// Selected read alignment
g2.setColor(Color.black);
}
else
{
// For a read with the same name as selected one.
g2.setColor(NON_SELECTED_READ_HIGHLIGHT_COLOUR);
}
g2.drawRect(xstart, ypos-BASE_HEIGHT, width, BASE_HEIGHT);
if(i < blocks.size()-1)
{
int nextStart =
(blocks.get(i+1).getReferenceStart() + offset - refSeqStart)*ALIGNMENT_PIX_PER_BASE;
g2.drawLine(xstart+width, ypos-(BASE_HEIGHT/2), nextStart, ypos-(BASE_HEIGHT/2));
}
g2.setColor(col1);
}
else if(i < blocks.size()-1)
{
refPos = block.getReferenceStart() + offset - refSeqStart;
int xstart = refPos*ALIGNMENT_PIX_PER_BASE;
int width = block.getLength()*ALIGNMENT_PIX_PER_BASE;
int nextStart =
(blocks.get(i+1).getReferenceStart() + offset - refSeqStart)*ALIGNMENT_PIX_PER_BASE;
g2.drawLine(xstart+width, ypos-(BASE_HEIGHT/2), nextStart, ypos-(BASE_HEIGHT/2));
}
if(lastMousePoint != null && blocks.size() > 0)
refPos = blocks.get(0).getReferenceStart()+offset-refSeqStart;
refPos = blocks.get(blocks.size()-1).getReferenceStart()+
blocks.get(blocks.size()-1).getLength()+offset-refSeqStart;
if(lastMousePoint.getY() > ypos-11 && lastMousePoint.getY() < ypos)
if(lastMousePoint.getX() > xstart &&
lastMousePoint.getX() < xend)
{
mouseOverSAMRecord = bamViewRecord;
if(insertions != null)
mouseOverInsertion = insertions.get((int)lastMousePoint.getX());
/**
* Colour bases on their mapping quality.
* @param g2
* @param baseQuality
*/
private void setColourByBaseQuality(Graphics2D g2, byte baseQuality)
{
if (baseQuality < 10)
g2.setColor(Color.blue);
else if (baseQuality < 20)
else if (baseQuality < 30)
else
g2.setColor(Color.black);
}
* Draw inferred size view.
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawLineView(Graphics2D g2, int seqLength, float pixPerBase, int start, int end)
drawSelectionRange(g2, pixPerBase,start, end, Color.PINK);
new BasicStroke (readLnHgt, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND);
if(isShowScale())
scaleHeight = 15;
else
scaleHeight = 0;
int baseAtStartOfView = getBaseAtStartOfView();
Rectangle r = jspView.getViewport().getViewRect();
BamViewRecord bamViewRecord = readsInView.get(i);
SAMRecord samRecord = bamViewRecord.sam;
BamViewRecord bamViewNextRecord = null;
if( !samRecord.getReadPairedFlag() || // read is not paired in sequencing
int ypos = getYPos(scaleHeight, samRecord.getReadString().length()); // (getHeight() - scaleHeight) - samRecord.getReadString().length();
if(ypos > r.getMaxY() || ypos < r.getMinY())
drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps, ydiff);
int ypos = getYPos(scaleHeight, Math.abs(samRecord.getInferredInsertSize()));
if( (ypos > r.getMaxY() || ypos < r.getMinY()) && ypos > 0 )
bamViewNextRecord = readsInView.get(++i);
samNextRecord = bamViewNextRecord.sam;
if(samRecord.getReadName().equals(samNextRecord.getReadName()))
if(samRecord.getAlignmentEnd() < samNextRecord.getAlignmentStart() &&
(samNextRecord.getAlignmentStart()-samRecord.getAlignmentEnd())*pixPerBase > 2.f)
g2.setColor(Color.LIGHT_GRAY);
int offset1 = getSequenceOffset(samRecord.getReferenceName());
int end1 = samRecord.getAlignmentEnd()+offset1-baseAtStartOfView;
int offset2 = getSequenceOffset(samNextRecord.getReferenceName());
int start2 = samNextRecord.getAlignmentStart()+offset2-baseAtStartOfView;
drawTranslucentLine(g2,
(int)(end1*pixPerBase), (int)(start2*pixPerBase), ypos);
g2.setColor(getColourByCoverageColour(bamViewRecord));
else if( (samRecord.getReadNegativeStrandFlag() && // strand of the query (1 for reverse)
samNextRecord.getReadNegativeStrandFlag()) ||
(!samRecord.getReadNegativeStrandFlag() &&
!samNextRecord.getReadNegativeStrandFlag()))
drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps, ydiff);
drawRead(g2, bamViewNextRecord, pixPerBase, ypos, baseAtStartOfView, getSNPs(samNextRecord), ydiff);
drawLoneRead(g2, bamViewRecord, ypos, pixPerBase, baseAtStartOfView, scaleHeight, snps, ydiff);
drawLoneRead(g2, bamViewRecord, ypos, pixPerBase, baseAtStartOfView, scaleHeight, snps, ydiff);
private int getYPos(int scaleHeight, int size)
{
if(!logScale)
else
{
int logInfSize = (int)( Math.log(size) * 100);
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/**
* Draw the reads as lines in vertical stacks. The reads are colour
* coded as follows:
*
* blue - reads are unique and are paired with a mapped mate
* black - reads are unique and are not paired or have an unmapped mate
* green - reads are duplicates
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*
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawStackView(Graphics2D g2,
final int seqLength,
final float pixPerBase,
final int start,
final int end)
drawSelectionRange(g2, pixPerBase,start, end, Color.PINK);
BasicStroke.CAP_BUTT,
BasicStroke.JOIN_MITER);
if(isShowScale())
scaleHeight = 15;
else
scaleHeight = 0;
int ypos = (getHeight() - scaleHeight);
int ydiff = (int) Math.round(1.5*readLnHgt);
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int maxEnd = 0;
SAMRecord samRecord = bamViewRecord.sam;
int offset = getSequenceOffset(samRecord.getReferenceName());
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int recordStart = samRecord.getAlignmentStart()+offset;
int recordEnd = samRecord.getAlignmentEnd()+offset;
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if(colourByStrandTag.isSelected())
{
if(samRecord.getAttribute("XS") == null)
g2.setColor(Color.BLACK);
else if( ((Character)samRecord.getAttribute("XS")).equals('+') )
g2.setColor(Color.BLUE);
else if( ((Character)samRecord.getAttribute("XS")).equals('-') )
g2.setColor(Color.RED);
else
g2.setColor(Color.BLACK);
}
else if(colourByCoverageColour.isSelected())
g2.setColor(getColourByCoverageColour(bamViewRecord));
else if(colourByReadGrp.isSelected())
g2.setColor(getReadGroupFrame().getReadGroupColour(readGroups, samRecord.getReadGroup()));
else if (samRecord.getDuplicateReadFlag())
g2.setColor(DARK_GREEN); // Duplicate
else if (!samRecord.getReadPairedFlag() || // read is not paired in sequencing
samRecord.getMateUnmappedFlag() ) // mate is unmapped ) // mate is unmapped
g2.setColor(Color.black);
else
g2.setColor(Color.blue);
if(maxEnd < recordStart || ypos < 0)
ypos = (getHeight() - scaleHeight)-ydiff;
maxEnd = recordEnd+2;
else
ypos = ypos-ydiff;
if(ypos > r.getMaxY() || ypos < r.getMinY())
drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps, ydiff);
/**
* Draw the reads as lines in vertical stacks. The reads are colour
* coded as follows:
*
* blue - reads are unique and are paired with a mapped mate
* black - reads are unique and are not paired or have an unmapped mate
* green - reads are duplicates
*
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawStrandStackView(Graphics2D g2,
int seqLength,
float pixPerBase,
int start,
int end)
{
drawSelectionRange(g2, pixPerBase,start, end, Color.PINK);
drawScale(g2, start, end, pixPerBase, ((getHeight()+scaleHeight)/2));
int ymid = (getHeight()/ 2);
drawStrand(g2, false, scaleHeight, ymid-(scaleHeight/2), -ydiff, pixPerBase);
drawStrand(g2, true, scaleHeight, ymid+(scaleHeight/2), ydiff, pixPerBase);
private void drawStrand(Graphics2D g2,
boolean isStrandNegative,
int scaleHeight,
int ymid,
int ystep,
int hgt = getHeight();
int ypos = (hgt - scaleHeight);
int baseAtStartOfView = getBaseAtStartOfView();
Rectangle r = jspView.getViewport().getViewRect();
SAMRecord samRecord = bamViewRecord.sam;
if( isNegativeStrand(samRecord, colourByStrandTag.isSelected()) == isStrandNegative )
final int offset = getSequenceOffset(samRecord.getReferenceName());
final int recordStart = samRecord.getAlignmentStart()+offset;
final int recordEnd = samRecord.getAlignmentEnd()+offset;
if(colourByStrandTag.isSelected())
if(samRecord.getAttribute("XS") == null)
g2.setColor(Color.BLACK);
else if( ((Character)samRecord.getAttribute("XS")).equals('+') )
g2.setColor(Color.BLUE);
else if( ((Character)samRecord.getAttribute("XS")).equals('-') )
g2.setColor(Color.RED);
else
g2.setColor(Color.BLACK);
}
else if(colourByCoverageColour.isSelected())
g2.setColor(getColourByCoverageColour(bamViewRecord));
else if(colourByReadGrp.isSelected())
g2.setColor(getReadGroupFrame().getReadGroupColour(readGroups, samRecord.getReadGroup()));
else if (samRecord.getDuplicateReadFlag())
g2.setColor(DARK_GREEN); // Duplicate
else if (!samRecord.getReadPairedFlag() || // read is not paired in sequencing
samRecord.getMateUnmappedFlag() ) // mate is unmapped
else
if(maxEnd < recordStart || ypos < 0 || ypos > hgt)
{
ypos = ymid + ystep;
maxEnd = recordEnd+2;
ypos = ypos + ystep;
if(ypos > r.getMaxY() || ypos < r.getMinY())
drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps, ystep);
* Draw paired reads as lines in a vertical stacks.
* @param g2
* @param seqLength
* @param pixPerBase
* @param start
* @param end
*/
private void drawPairedStackView(Graphics2D g2,
final int seqLength,
final float pixPerBase,
final int start,
final int end)
drawSelectionRange(g2, pixPerBase,start, end, Color.PINK);
if(isShowScale())
final Vector<PairedRead> pairedReads = new Vector<PairedRead>();
for(int i=0; i<readsInView.size(); i++)
{
BamViewRecord bamViewRecord = readsInView.get(i);
SAMRecord samRecord = bamViewRecord.sam;
if( !samRecord.getReadPairedFlag() || // read is not paired in sequencing
samRecord.getMateUnmappedFlag() ) // mate is unmapped
continue;
BamViewRecord bamViewNextRecord = null;
if(i < readsInView.size()-1)
{
bamViewNextRecord = readsInView.get(++i);
SAMRecord samNextRecord = bamViewNextRecord.sam;
final PairedRead pr = new PairedRead();
if(samRecord.getReadName().equals(samNextRecord.getReadName()) &&
isFromSameBamFile(bamViewRecord, bamViewNextRecord, bamList))
{
if(samRecord.getAlignmentStart() < samNextRecord.getAlignmentStart())
{
pr.sam1 = bamViewRecord;
pr.sam2 = bamViewNextRecord;
}
else
{
pr.sam2 = bamViewRecord;
pr.sam1 = bamViewNextRecord;
}
}
else
{
--i;
pr.sam1 = bamViewRecord;
pr.sam2 = null;
}
pairedReads.add(pr);
}
}
Collections.sort(pairedReads, new PairedReadComparator());
Stroke originalStroke = new BasicStroke (readLnHgt, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND);
g2.setStroke( new BasicStroke (1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND));
if(isShowScale())
scaleHeight = 15;
else
scaleHeight = 0;
int ypos = getHeight() - scaleHeight - ydiff;
int lastEnd = 0;
int baseAtStartOfView = getBaseAtStartOfView();
Rectangle r = jspView.getViewport().getViewRect();
if(pr.sam1.sam.getAlignmentStart() > lastEnd)
ypos = getHeight() - scaleHeight - ydiff;
if(pr.sam2 != null)
lastEnd = pr.sam2.sam.getAlignmentEnd();
lastEnd = pr.sam1.sam.getAlignmentEnd();
if(ypos > r.getMaxY() || ypos < r.getMinY())
g2.setStroke(originalStroke);
if( highlightSAMRecord != null &&
highlightSAMRecord.sam.getReadName().equals(pr.sam1.sam.getReadName()) )
g2.setColor(Color.black);
else
g2.setColor(Color.gray);
if(pr.sam2 != null)
{
if(!readsOverlap(pr.sam1.sam, pr.sam2.sam))
int offset1 = getSequenceOffset(pr.sam1.sam.getReferenceName());
int offset2 = getSequenceOffset(pr.sam2.sam.getReferenceName());
drawTranslucentJointedLine(g2,
(int)((pr.sam1.sam.getAlignmentEnd()+offset1-getBaseAtStartOfView())*pixPerBase),
(int)((pr.sam2.sam.getAlignmentStart()+offset2-getBaseAtStartOfView())*pixPerBase), ypos);
else if(!pr.sam1.sam.getMateUnmappedFlag() &&
pr.sam1.sam.getProperPairFlag() &&
pr.sam1.sam.getMateReferenceName().equals(pr.sam1.sam.getReferenceName()))
if(pr.sam1.sam.getAlignmentStart() > pr.sam1.sam.getMateAlignmentStart())
prStart = pr.sam1.sam.getMateAlignmentStart();
prEnd = pr.sam1.sam.getAlignmentStart();
prStart = pr.sam1.sam.getAlignmentEnd();
prEnd = pr.sam1.sam.getMateAlignmentStart();
int offset = getSequenceOffset(pr.sam1.sam.getReferenceName());
drawTranslucentJointedLine(g2,
(int)( (prStart+offset-getBaseAtStartOfView())*pixPerBase),
(int)( (prEnd +offset-getBaseAtStartOfView())*pixPerBase), ypos);
if(colourByCoverageColour.isSelected())
g2.setColor(getColourByCoverageColour(pr.sam1));
else if(colourByStrandTag.isSelected())
{
if( ((Character)pr.sam1.sam.getAttribute("XS")).equals('+') )
g2.setColor(Color.BLUE);
else if( ((Character)pr.sam1.sam.getAttribute("XS")).equals('-') )
g2.setColor(Color.RED);
else
g2.setColor(Color.BLACK);
}
else if(colourByReadGrp.isSelected())
g2.setColor(getReadGroupFrame().getReadGroupColour(readGroups, pr.sam1.sam.getReadGroup()));
else if( pr.sam2 != null &&
!( pr.sam1.sam.getReadNegativeStrandFlag() ^ pr.sam2.sam.getReadNegativeStrandFlag() ) )
g2.setColor(Color.red);
else
g2.setColor(Color.blue);
drawRead(g2, pr.sam1, pixPerBase, ypos, baseAtStartOfView, getSNPs(pr.sam1.sam), ydiff);
if(pr.sam2 != null)
drawRead(g2, pr.sam2, pixPerBase, ypos, baseAtStartOfView, getSNPs(pr.sam2.sam), ydiff);
/**
* Check if a record is on the negative strand. If the RNA strand specific
* checkbox is set then use the RNA strand.
* @param samRecord
* @param useStrandTag - strand specific tag
* @return
*/
protected static boolean isNegativeStrand(final SAMRecord samRecord,
final boolean useStrandTag)
if(useStrandTag)
{
if(samRecord.getAttribute("XS") == null)
return samRecord.getReadNegativeStrandFlag();
if( ((Character)samRecord.getAttribute("XS")).equals('+') )
return false;
else
return true;
}
return samRecord.getReadNegativeStrandFlag();
}
/**
* Check if two records are from the same BAM file
* @param sam1
* @param sam2
* @param bamList
* @return
*/
private boolean isFromSameBamFile(final BamViewRecord sam1,
final BamViewRecord sam2,
final List<String> bamList)
{
if(bamList == null || bamList.size()<2)
return true;
final short o1 = sam1.bamIndex;
final short o2 = sam2.bamIndex;
if(o1 != -1 && o2 != -1)
if( o1 != o2 )
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return false;
return true;
}
/**
* Check if two records overlap
* @param s1
* @param s2
* @return true id the two reads overlap
*/
private boolean readsOverlap(final SAMRecord s1,
final SAMRecord s2)
{
if( (s2.getAlignmentStart() >= s1.getAlignmentStart() &&
s2.getAlignmentStart() <= s1.getAlignmentEnd()) ||
(s2.getAlignmentEnd() >= s1.getAlignmentStart() &&
s2.getAlignmentEnd() <= s1.getAlignmentEnd()) )
return true;
if( (s1.getAlignmentStart() >= s2.getAlignmentStart() &&
s1.getAlignmentStart() <= s2.getAlignmentEnd()) ||
(s1.getAlignmentEnd() >= s2.getAlignmentStart() &&
s1.getAlignmentEnd() <= s2.getAlignmentEnd()) )
return true;
return false;
}
/**
* Draw the read coverage.
* @param g2
* @param start
* @param end
* @param pixPerBase
*/
private void drawCoverage(Graphics2D g2, int start, int end, float pixPerBase)
{
int scaleHeight = 0;
if(isShowScale())
{
drawScale(g2, start, end, pixPerBase, getHeight());
scaleHeight = 15;
}
int hgt = jspView.getVisibleRect().height-scaleHeight;
if(!cbCoverageStrandView.isSelected() && !coverageView.isPlotHeatMap())
try
{
int y = getHeight()-6-( (hgt* MAX_COVERAGE)/(coverageView.max/coverageView.windowSize) );
g2.setColor(Color.YELLOW);
// draw the threshold for the coverage max read cut-off
g2.fillRect(0, y, getWidth(), 8);
}
catch(Exception e){}
g2.translate(0, getJspView().getViewport().getViewPosition().y);
coverageView.drawSelectionRange(g2, pixPerBase, start, end, getHeight(), Color.PINK);
coverageView.drawMax(g2, coverageView.getMaxCoverage());
/**
* Draw a read that apparently has a read mate that is not in view.
* @param g2
* @param thisRead
* @param ypos
* @param pixPerBase
* @param originalStroke
* @param stroke
*/
private void drawLoneRead(Graphics2D g2, BamViewRecord bamViewRecord, int ypos,
float pixPerBase, int baseAtStartOfView, int scaleHeight, List<Integer> snps, int ydiff)
SAMRecord samRecord = bamViewRecord.sam;
boolean offTheTop = false;
int offset = getSequenceOffset(samRecord.getReferenceName());
int thisStart = samRecord.getAlignmentStart()+offset;
int thisEnd = thisStart + samRecord.getReadString().length() -1;
if(ypos <= 0)
{
offTheTop = true;
ypos = samRecord.getReadString().length();
}
{
offTheTop = true;
ypos = getHeight() - scaleHeight - 5;
}
Math.abs(samRecord.getMateAlignmentStart()-samRecord.getAlignmentEnd())*pixPerBase > 2.f)
{
g2.setColor(Color.LIGHT_GRAY);
if(samRecord.getAlignmentEnd() < samRecord.getMateAlignmentStart())
{
(int)((samRecord.getMateAlignmentStart()-getBaseAtStartOfView()+offset)*pixPerBase);
drawTranslucentLine(g2,
(int)((thisEnd-getBaseAtStartOfView())*pixPerBase), nextStart, ypos);
int nextStart =
(int)((samRecord.getMateAlignmentStart()-getBaseAtStartOfView()+offset)*pixPerBase);
drawTranslucentLine(g2,
(int)((thisStart-getBaseAtStartOfView())*pixPerBase), nextStart, ypos);
g2.setColor(getColourByCoverageColour(bamViewRecord));
tjc
committed
else if(samRecord.getReadNegativeStrandFlag() &&
samRecord.getMateNegativeStrandFlag()) // strand of the query (1 for reverse)
g2.setColor(Color.red);
else
g2.setColor(Color.blue);
drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps, ydiff);
/*if (isSNPs)
showSNPsOnReads(g2, samRecord, pixPerBase, ypos, offset);*/
private void drawScale(Graphics2D g2, int start, int end, float pixPerBase, int ypos)
g2.drawLine( 0, ypos-14,
(int)((end - getBaseAtStartOfView())*pixPerBase), ypos-14);
private void drawTicks(Graphics2D g2, int start, int end, float pixPerBase, int division, int ypos)
int markStart = (Math.round(start/division)*division);
if(markStart < 1)
markStart = 1;
g2.drawString(Integer.toString(m), x, ypos-1);
g2.drawLine((int)x, ypos-14,(int)x, ypos-11);
/**
* Draw a y-scale for inferred size (isize) of reads.
* @param g2
* @param xScaleHeight
*/
private void drawYScale(Graphics2D g2, int xScaleHeight)
int maxY = getPreferredSize().height-xScaleHeight;
if(logScale)
{
int start = 10;
int count = 0;
int ypos = getYPos(xScaleHeight, start);
while(ypos > 0 && count < 15 && start > 1)
{
g2.drawLine(0, ypos, 2, ypos);
g2.drawString(Integer.toString(start), 3, ypos);
start = start*5;
ypos = getYPos(xScaleHeight, start);
count++;
}
return;
}
for(int i=100; i<maxY; i+=100)
{
int ypos = getHeight()-i-xScaleHeight;
g2.drawLine(0, ypos, 2, ypos);
g2.drawString(Integer.toString(i), 3, ypos);
/**
* Draw a given read.
* @param g2
* @param thisRead
* @param pixPerBase
* @param ypos
* @param snps
final BamViewRecord bamViewRecord,
final int baseAtStartOfView,
SAMRecord thisRead = bamViewRecord.sam;
int offset = getSequenceOffset(thisRead.getReferenceName());
int thisStart = thisRead.getAlignmentStart()+offset-baseAtStartOfView;
int thisEnd = thisRead.getAlignmentEnd()+offset-baseAtStartOfView;
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if(highlightSAMRecord != null &&
highlightSAMRecord.sam.getReadName().equals(thisRead.getReadName()))
{
Stroke originalStroke = g2.getStroke();
Stroke stroke =
new BasicStroke (readLnHgt*1.6f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND);
g2.setStroke(stroke);
Color c = g2.getColor();
if (isThisBamRecordHighlighted(bamViewRecord) )
{
// Selected read alignment
g2.setColor(Color.black);
}
else
{
// For a read with the same name as selected one.
g2.setColor(NON_SELECTED_READ_HIGHLIGHT_COLOUR);
}
g2.drawLine((int)( thisStart * pixPerBase), ypos,
(int)( thisEnd * pixPerBase), ypos);
g2.setColor(c);
g2.setStroke(originalStroke);
}
if(thisRead.getCigar().getCigarElements().size() == 1)
g2.drawLine((int)( thisStart * pixPerBase), ypos,
(int)( thisEnd * pixPerBase), ypos);
else
{
List<AlignmentBlock> blocks = thisRead.getAlignmentBlocks();
Color c = g2.getColor();
int lastEnd = 0;
for(int i=0; i<blocks.size(); i++)
{
AlignmentBlock block = blocks.get(i);
int blockStart = block.getReferenceStart()+offset-baseAtStartOfView;
int blockEnd = blockStart + block.getLength() - 1;
g2.drawLine((int)( blockStart * pixPerBase), ypos,
(int)( blockEnd * pixPerBase), ypos);
if(i > 0 && blockStart != lastEnd)
{
g2.setColor(Color.gray);
g2.drawLine((int)( blockStart * pixPerBase), ypos,
(int)( lastEnd * pixPerBase), ypos);
g2.setColor(c);
}
lastEnd = blockEnd;
}
}
drawArrow(g2, thisRead, thisStart, thisEnd, pixPerBase, ypos, ydiff);
// test if the mouse is over this read
if(lastMousePoint.getY() > ypos-(Math.abs(ydiff)/2) && lastMousePoint.getY() < ypos+(Math.abs(ydiff)/2))
{
if(lastMousePoint.getX() > thisStart * pixPerBase &&
lastMousePoint.getX() < thisEnd * pixPerBase)
{
mouseOverSAMRecord = bamViewRecord;
}
}
if (isSNPs && snps != null && snps.size() > 0)
showSNPsOnReads(snps, g2, pixPerBase, ypos);
/**
* Draw arrow on the read to indicate orientation.
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