/* BamView * * 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. * */ package uk.ac.sanger.artemis.components.alignment; 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.Dimension; import java.awt.FlowLayout; import java.awt.FontMetrics; import java.awt.Graphics; import java.awt.Graphics2D; import java.awt.GridBagConstraints; import java.awt.GridBagLayout; import java.awt.Insets; import java.awt.Point; import java.awt.Rectangle; import java.awt.Stroke; 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.event.MouseAdapter; import java.awt.event.MouseEvent; import java.awt.event.MouseMotionListener; import java.awt.image.BufferedImage; import java.io.BufferedReader; import java.io.File; import java.io.FileOutputStream; import java.io.IOException; import java.io.InputStream; import java.io.InputStreamReader; import java.lang.management.ManagementFactory; import java.lang.management.MemoryMXBean; import java.lang.reflect.Field; import java.net.URL; import java.util.ArrayList; import java.util.Collections; import java.util.Enumeration; import java.util.HashMap; import java.util.Hashtable; import java.util.List; import java.util.Map; import java.util.Vector; import java.util.concurrent.CountDownLatch; import java.util.concurrent.ExecutorService; import java.util.concurrent.Executors; import javax.swing.BorderFactory; import javax.swing.Box; import javax.swing.BoxLayout; import javax.swing.ButtonGroup; import javax.swing.ImageIcon; import javax.swing.JButton; import javax.swing.JCheckBox; import javax.swing.JCheckBoxMenuItem; import javax.swing.JComponent; import javax.swing.JFrame; import javax.swing.JLabel; import javax.swing.JMenu; import javax.swing.JMenuBar; import javax.swing.JMenuItem; import javax.swing.JOptionPane; import javax.swing.JPanel; import javax.swing.JPopupMenu; import javax.swing.JRadioButton; import javax.swing.JScrollBar; import javax.swing.JScrollPane; import javax.swing.JSeparator; import javax.swing.JTextField; import javax.swing.SwingUtilities; import javax.swing.UIManager; import javax.swing.border.Border; import javax.swing.border.EmptyBorder; 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.Options; import uk.ac.sanger.artemis.Selection; import uk.ac.sanger.artemis.SelectionChangeEvent; import uk.ac.sanger.artemis.SelectionChangeListener; import uk.ac.sanger.artemis.SimpleEntryGroup; 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.EntryFileDialog; import uk.ac.sanger.artemis.components.FeatureDisplay; import uk.ac.sanger.artemis.components.FileViewer; import uk.ac.sanger.artemis.components.IndexReferenceEvent; import uk.ac.sanger.artemis.components.MessageDialog; 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.io.EntryInformation; import uk.ac.sanger.artemis.io.Range; import uk.ac.sanger.artemis.sequence.Bases; 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; import uk.ac.sanger.artemis.util.OutOfRangeException; public class BamView extends JPanel implements DisplayAdjustmentListener, SelectionChangeListener { private static final long serialVersionUID = 1L; private List<BamViewRecord> readsInView; private Hashtable<String, SAMFileReader> samFileReaderHash = new Hashtable<String, SAMFileReader>(); private HashMap<String, Integer> seqLengths = new HashMap<String, Integer>(); private HashMap<String, Integer> offsetLengths; private Vector<String> seqNames = new Vector<String>(); protected List<String> bamList; protected List<Short> hideBamList = new Vector<Short>(); private SAMRecordPredicate samRecordFlagPredicate; private SAMRecordMapQPredicate samRecordMapQPredicate; private SAMRecordFilter filterFrame; private Bases bases; private JScrollPane jspView; private JScrollBar scrollBar; private SequenceComboBox combo; private boolean isOrientation = false; private boolean isSingle = false; private boolean isSNPs = false; private boolean isCoverage = false; private boolean isSNPplot = false; private EntryEdit entry_edit; private FeatureDisplay feature_display; private Selection selection; private JPanel mainPanel = new JPanel(); private CoveragePanel coveragePanel; private SnpPanel snpPanel; protected boolean logScale = false; private Ruler ruler; private int nbasesInView; private int startBase = -1; private int endBase = -1; private int laststart; private int lastend; private boolean asynchronous = true; 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 colourByCoverageColour = new JCheckBoxMenuItem("Coverage Plot Colours"); private JCheckBoxMenuItem baseQualityColour = new JCheckBoxMenuItem("Base Quality"); private JCheckBoxMenuItem markInsertions = new JCheckBoxMenuItem("Mark Insertions", true); 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)$"; /** Used to colour the frames. */ 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 Point lastMousePoint = null; private BamViewRecord mouseOverSAMRecord = null; private BamViewRecord highlightSAMRecord = null; private String mouseOverInsertion; // record of where a mouse drag starts protected int dragStart = -1; private static int MAX_BASES = 26000; private int maxHeight = 800; private boolean concatSequences = false; private int ALIGNMENT_PIX_PER_BASE; private int BASE_HEIGHT; private JPopupMenu popup; 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, String reference, int nbasesInView, final FeatureDisplay feature_display, final Bases bases, final JPanel containerPanel, final JFrame frame) { super(); setBackground(Color.white); this.bamList = bamList; this.nbasesInView = nbasesInView; 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)); if(reference != null) { System.setProperty("reference", reference); // for CRAM EntryGroup entryGroup = new SimpleEntryGroup(); 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(); Enumeration<Object> keys = UIManager.getDefaults().keys(); 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()); FontMetrics fm = getFontMetrics(getFont()); ALIGNMENT_PIX_PER_BASE = fm.charWidth('M'); BASE_HEIGHT = fm.getMaxAscent(); selection = new Selection(null); MultiLineToolTipUI.initialize(); setToolTipText(""); buttonGroup.add(cbStackView); buttonGroup.add(cbPairedStackView); buttonGroup.add(cbStrandStackView); buttonGroup.add(cbIsizeStackView); buttonGroup.add(cbCoverageView); buttonGroup.add(cbCoverageStrandView); buttonGroup.add(cbCoverageHeatMap); addMouseListener(new PopupListener()); 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); addBamToPanel(frame); } public String getToolTipText() { 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() + "\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()); } else { bamIndexFile = new File(bam + ".bai"); if(!bamIndexFile.exists()) { final File cramIndexFile = new File(bam + ".crai"); if(cramIndexFile.exists()) return cramIndexFile; } } return bamIndexFile; } /** * 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); File bamIndexFile = getBamIndexFile(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); } } final SAMFileReader samFileReader; if(feature_display != null && bam.endsWith("cram")) { 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"); 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")) { File bamFile = new File(bam); 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); return samFileReader; } private void readHeaderPicard() throws IOException { 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()); } } 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(); } } } /** * Read a SAM or BAM file. * @throws IOException */ private void readFromBamPicard(int start, int end, short bamIndex, float pixPerBase) throws IOException { // Open the input file. Automatically detects whether input is SAM or BAM // and delegates to a reader implementation for the appropriate format. final String bam = bamList.get(bamIndex); final SAMFileReader inputSam = getSAMFileReader(bam); //final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile); if(isConcatSequences()) { for(String seq: seqNames) { int sLen = seqLengths.get(seq); int offset = getSequenceOffset(seq); 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); } //inputSam.close(); } /** * 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 checkMemAfter = 8000; final int seqOffset = getSequenceOffset(refName); final int offset = seqOffset- getBaseAtStartOfView(); final boolean isCoverageView = isCoverageView(pixPerBase); int cnt = 0; 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 { while ( it.hasNext() ) { try { cnt++; SAMRecord samRecord = it.next(); 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) 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); if(isCoverage) coveragePanel.addRecord(samRecord, offset, bam); if(isSNPplot) snpPanel.addRecord(samRecord, seqOffset); if(!isCoverageView) readsInView.add(new BamViewRecord(samRecord, bamIndex)); } } if(cnt > checkMemAfter) { 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(); } } private int getSequenceLength() { if(isConcatSequences()) { int len = 0; for(String seq: seqNames) len += seqLengths.get(seq); 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(!isConcatSequences()) return 0; if(offsetLengths == null) { /* offsetLengths = new Hashtable<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)); }*/ 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); } }); //concatSequences = false; 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); } /** * Override */ protected void paintComponent(Graphics g) { super.paintComponent(g); Graphics2D g2 = (Graphics2D)g; mouseOverSAMRecord = null; final int seqLength = getSequenceLength(); int start; int end; if(startBase > 0) start = startBase; else start = getBaseAtStartOfView(); 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(); } final float pixPerBase = getPixPerBaseByWidth(); boolean changeToStackView = false; MemoryMXBean memory = ManagementFactory.getMemoryMXBean(); if(laststart != start || lastend != end || CoveragePanel.isRedraw()) { if(isCoverageView(pixPerBase)) 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) { cbStackView.setSelected(true); 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()); } } } laststart = start; lastend = end; if(showBaseAlignment) drawBaseAlignment(g2, seqLength, pixPerBase, start, end); else { 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(waitingFrame.isVisible()) waitingFrame.hideFrame(); if(changeToStackView) { popFrame.show( "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.", mainPanel, 15000); } } private float getPixPerBaseByWidth() { return (float)mainPanel.getWidth() / (float)nbasesInView; } private int getMaxBasesInPanel(int seqLength) { if(feature_display == null) return seqLength+nbasesInView/3; else return seqLength+nbasesInView; } /** * 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, int end) { ruler.start = start; ruler.end = end; int ypos = 0; String refSeq = null; int refSeqStart = start; end = start + ( mainPanel.getWidth() * ALIGNMENT_PIX_PER_BASE ); if(bases != null) { try { int seqEnd = end+2; 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)); boolean drawn[] = new boolean[readsInView.size()]; for(int i=0; i<readsInView.size(); i++) drawn[i] = false; Rectangle r = jspView.getViewport().getViewRect(); int nreads = readsInView.size(); for (int i = 0; i < nreads; i++) { try { if (!drawn[i]) { ypos += 11; 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(); if (thisEnd == 0) thisEnd = thisRead.sam.getAlignmentStart() + thisRead.sam.getReadLength(); for (int j = i + 1; j < nreads; j++) { if (!drawn[j]) { 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(); if (thisEnd == 0) 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(); } } if(ypos > getHeight()) { Dimension d = getPreferredSize(); d.setSize(getPreferredSize().getWidth(), ypos); setPreferredSize(d); revalidate(); } } /** * Draw the query sequence * @param g2 * @param read * @param pixPerBase * @param ypos */ private void drawSequence(Graphics2D g2, 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 g2.setColor(Color.black); else g2.setColor(Color.blue); final Color col = g2.getColor(); int xpos; int len = 0; int refPos = 0; String readSeq = samRecord.getReadString(); 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(); len += block.getLength(); 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) setColourByBaseQuality(g2, phredQuality[readPos]); if(isSNPs && refSeq != null && refPos > 0 && refPos < refSeq.length()) { if(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>(); g2.setColor(DEEP_PINK); int xscreen = (refPos+1)*ALIGNMENT_PIX_PER_BASE; insertions.put(xscreen, (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); } } // highlight if(highlightSAMRecord != null && highlightSAMRecord.sam.getReadName().equals(samRecord.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); 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; int xstart = refPos*ALIGNMENT_PIX_PER_BASE; refPos = blocks.get(blocks.size()-1).getReferenceStart()+ blocks.get(blocks.size()-1).getLength()+offset-refSeqStart; int xend = (refPos+len)*ALIGNMENT_PIX_PER_BASE; 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) g2.setColor(DARK_GREEN); else if (baseQuality < 30) g2.setColor(DARK_ORANGE); 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); if(isShowScale()) drawScale(g2, start, end, pixPerBase, getHeight()); final Stroke stroke = new BasicStroke (1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND); g2.setStroke(stroke); final int scaleHeight; if(isShowScale()) scaleHeight = 15; else scaleHeight = 0; int baseAtStartOfView = getBaseAtStartOfView(); Rectangle r = jspView.getViewport().getViewRect(); for(int i=0; i<readsInView.size(); i++) { BamViewRecord bamViewRecord = readsInView.get(i); SAMRecord samRecord = bamViewRecord.sam; BamViewRecord bamViewNextRecord = null; SAMRecord samNextRecord = null; List<Integer> snps = getSNPs(samRecord); if( !samRecord.getReadPairedFlag() || // read is not paired in sequencing samRecord.getMateUnmappedFlag() ) // mate is unmapped { if(isSingle) { int ypos = getYPos(scaleHeight, samRecord.getReadString().length()); // (getHeight() - scaleHeight) - samRecord.getReadString().length(); if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; g2.setColor(Color.black); drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps); } continue; } int ypos = getYPos(scaleHeight, Math.abs(samRecord.getInferredInsertSize())); if( (ypos > r.getMaxY() || ypos < r.getMinY()) && ypos > 0 ) continue; if(i < readsInView.size()-1) { bamViewNextRecord = readsInView.get(++i); samNextRecord = bamViewNextRecord.sam; if(samRecord.getReadName().equals(samNextRecord.getReadName())) { // draw connection between paired reads 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); } if(colourByCoverageColour.isSelected()) g2.setColor(getColourByCoverageColour(bamViewRecord)); else if( (samRecord.getReadNegativeStrandFlag() && // strand of the query (1 for reverse) samNextRecord.getReadNegativeStrandFlag()) || (!samRecord.getReadNegativeStrandFlag() && !samNextRecord.getReadNegativeStrandFlag())) g2.setColor(Color.red); else g2.setColor(Color.blue); drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps); drawRead(g2, bamViewNextRecord, pixPerBase, ypos, baseAtStartOfView, getSNPs(samNextRecord)); } else { drawLoneRead(g2, bamViewRecord, ypos, pixPerBase, baseAtStartOfView, scaleHeight, snps); i--; } } else { drawLoneRead(g2, bamViewRecord, ypos, pixPerBase, baseAtStartOfView, scaleHeight, snps); } } drawYScale(g2, scaleHeight); } private int getYPos(int scaleHeight, int size) { if(!logScale) return (getHeight() - scaleHeight) - size; else { int logInfSize = (int)( Math.log(size) * 100); return (getHeight() - scaleHeight) - logInfSize; } } /** * 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 drawStackView(Graphics2D g2, final int seqLength, final float pixPerBase, final int start, final int end) { drawSelectionRange(g2, pixPerBase,start, end, Color.PINK); if(isShowScale()) drawScale(g2, start, end, pixPerBase, getHeight()); final BasicStroke stroke = new BasicStroke( 1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_MITER); g2.setStroke(stroke); final int scaleHeight; if(isShowScale()) scaleHeight = 15; else scaleHeight = 0; int ypos = (getHeight() - scaleHeight); int ydiff = 2; if(isOrientation) ydiff= 4; int maxEnd = 0; int lstStart = 0; int lstEnd = 0; final int baseAtStartOfView = getBaseAtStartOfView(); g2.setColor(Color.blue); final Rectangle r = jspView.getViewport().getViewRect(); for(BamViewRecord bamViewRecord: readsInView) { SAMRecord samRecord = bamViewRecord.sam; int offset = getSequenceOffset(samRecord.getReferenceName()); int recordStart = samRecord.getAlignmentStart()+offset; int recordEnd = samRecord.getAlignmentEnd()+offset; List<Integer> snps = getSNPs(samRecord); if(colourByCoverageColour.isSelected() || lstStart != recordStart || lstEnd != recordEnd || snps != null) { if(colourByCoverageColour.isSelected()) g2.setColor(getColourByCoverageColour(bamViewRecord)); 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; } else g2.setColor(DARK_GREEN); if(snps != null) lstStart = -1; else { lstStart = recordStart; lstEnd = recordEnd; } if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps); } } /** * 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); final BasicStroke stroke = new BasicStroke( 1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_MITER); final int scaleHeight = 15; drawScale(g2, start, end, pixPerBase, ((getHeight()+scaleHeight)/2)); int ymid = (getHeight()/ 2); int ydiff = 2; if(isOrientation) ydiff= 4; // positive strand drawStrand(g2, false, scaleHeight, ymid-(scaleHeight/2), -ydiff, pixPerBase, stroke); // negative strand drawStrand(g2, true, scaleHeight, ymid+(scaleHeight/2), ydiff, pixPerBase, stroke); } private void drawStrand(Graphics2D g2, boolean isStrandNegative, int scaleHeight, int ymid, int ystep, float pixPerBase, Stroke stroke) { int hgt = getHeight(); int ypos = (hgt - scaleHeight); int maxEnd = 0; int lstStart = 0; int lstEnd = 0; int baseAtStartOfView = getBaseAtStartOfView(); g2.setColor(Color.blue); Rectangle r = jspView.getViewport().getViewRect(); for(BamViewRecord bamViewRecord: readsInView) { SAMRecord samRecord = bamViewRecord.sam; if( samRecord.getReadNegativeStrandFlag() == isStrandNegative ) { final int offset = getSequenceOffset(samRecord.getReferenceName()); final int recordStart = samRecord.getAlignmentStart()+offset; final int recordEnd = samRecord.getAlignmentEnd()+offset; List<Integer> snps = getSNPs(samRecord); if(colourByCoverageColour.isSelected() || lstStart != recordStart || lstEnd != recordEnd || snps != null) { if(colourByCoverageColour.isSelected()) g2.setColor(getColourByCoverageColour(bamViewRecord)); else if (!samRecord.getReadPairedFlag() || // read is not paired in sequencing samRecord.getMateUnmappedFlag() ) // mate is unmapped g2.setColor(Color.black); else g2.setColor(Color.blue); if(maxEnd < recordStart || ypos < 0 || ypos > hgt) { ypos = ymid + ystep; maxEnd = recordEnd+2; } else ypos = ypos + ystep; } else g2.setColor(DARK_GREEN); if(snps != null) lstStart = -1; else { lstStart = recordStart; lstEnd = recordEnd; } if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; drawRead(g2, bamViewRecord, pixPerBase, ypos, baseAtStartOfView, snps); } } } /** * 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()) drawScale(g2, start, end, pixPerBase, getHeight()); 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 (1.0f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND); Stroke stroke = new BasicStroke (1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND); g2.setStroke(stroke); final int scaleHeight; if(isShowScale()) scaleHeight = 15; else scaleHeight = 0; int ydiff = 3; if(isOrientation) ydiff= 5; int ypos = getHeight() - scaleHeight - ydiff; int lastEnd = 0; int baseAtStartOfView = getBaseAtStartOfView(); Rectangle r = jspView.getViewport().getViewRect(); for(PairedRead pr: pairedReads) { if(pr.sam1.sam.getAlignmentStart() > lastEnd) { ypos = getHeight() - scaleHeight - ydiff; if(pr.sam2 != null) lastEnd = pr.sam2.sam.getAlignmentEnd(); else lastEnd = pr.sam1.sam.getAlignmentEnd(); } else ypos = ypos - ydiff; if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; 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())) { final int prStart, prEnd; if(pr.sam1.sam.getAlignmentStart() > pr.sam1.sam.getMateAlignmentStart()) { prStart = pr.sam1.sam.getMateAlignmentStart(); prEnd = pr.sam1.sam.getAlignmentStart(); } else { 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( 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)); if(pr.sam2 != null) drawRead(g2, pr.sam2, pixPerBase, ypos, baseAtStartOfView, getSNPs(pr.sam2.sam)); } } /** * 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 ) 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.draw(g2, getWidth(), hgt, hideBamList); if(!coverageView.isPlotHeatMap()) coverageView.drawMax(g2); } /** * 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) { 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(); } if(samRecord.getInferredInsertSize() == 0) { offTheTop = true; ypos = getHeight() - scaleHeight - 5; } if(samRecord.getInferredInsertSize() != 0 && Math.abs(samRecord.getMateAlignmentStart()-samRecord.getAlignmentEnd())*pixPerBase > 2.f) { g2.setColor(Color.LIGHT_GRAY); if(samRecord.getAlignmentEnd() < samRecord.getMateAlignmentStart()) { int nextStart = (int)((samRecord.getMateAlignmentStart()-getBaseAtStartOfView()+offset)*pixPerBase); drawTranslucentLine(g2, (int)((thisEnd-getBaseAtStartOfView())*pixPerBase), nextStart, ypos); } else { int nextStart = (int)((samRecord.getMateAlignmentStart()-getBaseAtStartOfView()+offset)*pixPerBase); drawTranslucentLine(g2, (int)((thisStart-getBaseAtStartOfView())*pixPerBase), nextStart, ypos); } } if(colourByCoverageColour.isSelected()) g2.setColor(getColourByCoverageColour(bamViewRecord)); else if(offTheTop) g2.setColor(DARK_ORANGE); 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); /*if (isSNPs) showSNPsOnReads(g2, samRecord, pixPerBase, ypos, offset);*/ } private void drawScale(Graphics2D g2, int start, int end, float pixPerBase, int ypos) { g2.setColor(Color.black); g2.drawLine( 0, ypos-14, (int)((end - getBaseAtStartOfView())*pixPerBase), ypos-14); int interval = end-start; if(interval > 256000) drawTicks(g2, start, end, pixPerBase, 512000, ypos); else if(interval > 64000) drawTicks(g2, start, end, pixPerBase, 12800, ypos); else if(interval > 16000) drawTicks(g2, start, end, pixPerBase, 3200, ypos); else if(interval > 4000) drawTicks(g2, start, end, pixPerBase, 800, ypos); else if(interval > 1000) drawTicks(g2, start, end, pixPerBase, 400, ypos); else drawTicks(g2, start, end, pixPerBase, 100, ypos); } 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; int sm = markStart-(division/2); float x; if(sm > start) { x = (sm-getBaseAtStartOfView())*pixPerBase; g2.drawLine((int)x, ypos-14,(int)x, ypos-12); } for(int m=markStart; m<end; m+=division) { x = (m-getBaseAtStartOfView())*pixPerBase; g2.drawString(Integer.toString(m), x, ypos-1); g2.drawLine((int)x, ypos-14,(int)x, ypos-11); sm = m+(division/2); if(sm < end) { x = (sm-getBaseAtStartOfView())*pixPerBase; g2.drawLine((int)x, ypos-14,(int)x, ypos-12); } if(m == 1) m = 0; } } /** * Draw a y-scale for inferred size (isize) of reads. * @param g2 * @param xScaleHeight */ private void drawYScale(Graphics2D g2, int xScaleHeight) { g2.setColor(Color.black); 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 baseAtStartOfView * @param snps */ private void drawRead(Graphics2D g2, final BamViewRecord bamViewRecord, final float pixPerBase, final int ypos, final int baseAtStartOfView, final List<Integer> snps) { SAMRecord thisRead = bamViewRecord.sam; int offset = getSequenceOffset(thisRead.getReferenceName()); int thisStart = thisRead.getAlignmentStart()+offset-baseAtStartOfView; int thisEnd = thisRead.getAlignmentEnd()+offset-baseAtStartOfView; if(highlightSAMRecord != null && highlightSAMRecord.sam.getReadName().equals(thisRead.getReadName())) { Stroke originalStroke = g2.getStroke(); Stroke stroke = new BasicStroke (3.f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND); g2.setStroke(stroke); Color c = g2.getColor(); g2.setColor(Color.black); 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; } } if(isOrientation) drawArrow(g2, thisRead, thisStart, thisEnd, pixPerBase, ypos); // test if the mouse is over this read if(lastMousePoint != null) { if(lastMousePoint.getY()+2 > ypos && lastMousePoint.getY()-2 < ypos) if(lastMousePoint.getX() > thisStart * pixPerBase && lastMousePoint.getX() < thisEnd * pixPerBase) { mouseOverSAMRecord = bamViewRecord; } } if (isSNPs && snps != null) showSNPsOnReads(snps, g2, pixPerBase, ypos); } /** * Draw arrow on the read to indicate orientation. * @param g2 * @param thisRead * @param thisStart * @param thisEnd * @param pixPerBase * @param ypos */ private void drawArrow(Graphics2D g2, SAMRecord thisRead, int thisStart, int thisEnd, float pixPerBase, int ypos) { if(thisRead.getReadNegativeStrandFlag()) { int apos = ypos + 2; g2.drawLine((int)( (thisStart+5) * pixPerBase), apos, (int)( thisStart * pixPerBase), ypos); } else { int apos = ypos - 2; g2.drawLine((int)( (thisEnd-5) * pixPerBase), apos, (int)( thisEnd * pixPerBase), ypos); } } /** * Highlight a selected range * @param g2 * @param pixPerBase * @param start * @param end */ private void drawSelectionRange(Graphics2D g2, float pixPerBase, int start, int end, Color c) { if(getSelection() != null) { Range selectedRange = getSelection().getSelectionRange(); if(selectedRange != null) { int rangeStart = selectedRange.getStart(); int rangeEnd = selectedRange.getEnd(); if(end < rangeStart || start > rangeEnd) return; int x = (int) (pixPerBase*(rangeStart-getBaseAtStartOfView())); int width = (int) (pixPerBase*(rangeEnd-rangeStart+1)); g2.setColor(c); g2.fillRect(x, 0, width, getHeight()); } } } /** * Draw a translucent line * @param g2 * @param start * @param end * @param ypos */ private void drawTranslucentLine(Graphics2D g2, int start, int end, int ypos) { Composite origComposite = g2.getComposite(); g2.setComposite(translucent); g2.drawLine(start, ypos, end, ypos); g2.setComposite(origComposite); } /** * Draw a translucent line * @param g2 * @param start * @param end * @param ypos */ private void drawTranslucentJointedLine(Graphics2D g2, int start, int end, int ypos) { Composite origComposite = g2.getComposite(); g2.setComposite(translucent); int mid = (int) ((end-start)/2.f)+start; //g2.drawLine(start, ypos, end, ypos); g2.drawLine(start, ypos, mid, ypos-5); g2.drawLine(mid, ypos-5, end, ypos); g2.setComposite(origComposite); } /** * Display the SNPs for the given read. * @param snps * @param g2 * @param pixPerBase * @param ypos */ private void showSNPsOnReads(final List<Integer> snps, final Graphics2D g2, float pixPerBase, int ypos) { g2.setColor(Color.red); for(int pos: snps) g2.drawLine((int) (pos * pixPerBase), ypos + 2, (int) (pos * pixPerBase), ypos - 2); } /** * Get the SNP positions * @param samRecord */ private List<Integer> getSNPs(final SAMRecord samRecord) { if(!isSNPs) // return null if not displaying SNPs return null; int rbeg = samRecord.getAlignmentStart(); int rend = samRecord.getAlignmentEnd(); int offset = getSequenceOffset(samRecord.getReferenceName()); ArrayList<Integer> snps = null; // use alignment blocks of the contiguous alignment of // subsets of read bases to a reference sequence try { final char[] refSeq = bases.getSubSequenceC( new Range(rbeg+offset, rend+offset), Bases.FORWARD); final byte[] readSeq = samRecord.getReadBases(); offset = offset - getBaseAtStartOfView(); final List<AlignmentBlock> blocks = samRecord.getAlignmentBlocks(); for(AlignmentBlock block: blocks) { int readStart = block.getReadStart(); int refStart = block.getReferenceStart(); int len = block.getLength(); for(int j=0; j<len; j++) { int readPos = readStart-1+j; int refPos = refStart+j; if (Character.toUpperCase(refSeq[refPos-rbeg]) != readSeq[readPos]) { if(snps == null) snps = new ArrayList<Integer>(); snps.add(refPos+offset); } } } } catch (OutOfRangeException e) { System.err.println(samRecord.getReadName()+" "+e.getMessage()); } return snps; } /** * Add the alignment view to the supplied <code>JPanel</code> in * a <code>JScrollPane</code>. * @param mainPanel panel to add the alignment to * @param frame * @param autohide automatically hide the top panel containing the buttons * @param feature_display */ private void addBamToPanel(final JFrame frame) { final JComponent topPanel = bamTopPanel(frame); mainPanel.setPreferredSize(new Dimension(900, 400)); setDisplay(1, nbasesInView, null); mainPanel.setLayout(new BorderLayout()); if(topPanel instanceof JPanel) mainPanel.add(topPanel, BorderLayout.NORTH); mainPanel.add(jspView, BorderLayout.CENTER); JPanel bottomPanel = new JPanel(new BorderLayout()); coveragePanel = new CoveragePanel(this); bottomPanel.add(coveragePanel, BorderLayout.CENTER); // snpPanel = new SnpPanel(this, bases); bottomPanel.add(snpPanel, BorderLayout.NORTH); if(feature_display == null) { scrollBar = new JScrollBar(JScrollBar.HORIZONTAL, 1, nbasesInView, 1, getMaxBasesInPanel(getSequenceLength())); scrollBar.setUnitIncrement(nbasesInView/20); scrollBar.addAdjustmentListener(new AdjustmentListener() { public void adjustmentValueChanged(AdjustmentEvent e) { repaint(); if(isSNPplot) snpPanel.repaint(); if(isCoverage) coveragePanel.repaint(); } }); bottomPanel.add(scrollBar, BorderLayout.SOUTH); } else { if(!isConcatSequences()) { int seqLen = seqLengths.get((String) combo.getSelectedItem()); int artemisSeqLen = feature_display.getSequenceLength(); if(seqLen != artemisSeqLen) { int newIndex = -1; for(int i=0; i<seqNames.size(); i++) { if(seqLengths.get(seqNames.get(i)) == artemisSeqLen) { // this looks like the correct sequence combo.setSelectedIndex(i); newIndex = i; } } if(!Options.isBlackBeltMode()) { String label[] = { "The length of the sequence loaded does not match the length of", "the default reference sequence in the BAM ("+seqNames.get(0)+").", (newIndex == -1 ? "" : "The length does match the reference "+ seqNames.get(newIndex)+" so this has been set as the default.") }; new NonModalDialog(frame, label); } } } } mainPanel.add(bottomPanel, BorderLayout.SOUTH); coveragePanel.setPreferredSize(new Dimension(900, 100)); coveragePanel.setVisible(false); snpPanel.setPreferredSize(new Dimension(900, 100)); snpPanel.setVisible(false); mainPanel.revalidate(); jspView.getVerticalScrollBar().setValue( jspView.getVerticalScrollBar().getMaximum()); } private void addToViewMenu(final short thisBamIndex) { final File f = new File(bamList.get(thisBamIndex)); final JCheckBoxMenuItem cbBam = new JCheckBoxMenuItem( f.getName(), getImageIcon(getColourByCoverageColour(thisBamIndex)), true); bamFilesMenu.add(cbBam); cbBam.addItemListener(new ItemListener() { public void itemStateChanged(ItemEvent e) { if(cbBam.isSelected()) hideBamList.remove(new Short(thisBamIndex)); else hideBamList.add(new Short(thisBamIndex)); laststart = -1; repaint(); } }); } /** * Refresh the colour of the icons used to identify the * BAM files. */ protected void refreshColourOfBamMenu() { final Component cs[] = bamFilesMenu.getMenuComponents(); for(Component c : cs) { if(c instanceof JCheckBoxMenuItem) { final JCheckBoxMenuItem cbBam = (JCheckBoxMenuItem) c; final Color col = getColorByJCheckBoxMenuItem(cbBam); if(col != null) cbBam.setIcon(getImageIcon(col)); } } } protected Color getColorByJCheckBoxMenuItem(JCheckBoxMenuItem cbBam) { final String bam = cbBam.getText(); for(short i=0; i<bamList.size(); i++) { final File f = new File(bamList.get(i)); if(f.getName().equals(bam)) return getColourByCoverageColour(i); } return null; } /** * Create an icon of a box using the given colour. * @param c * @return */ protected ImageIcon getImageIcon(Color c) { BufferedImage image = (BufferedImage)this.createImage(10, 10); Graphics2D g2 = image.createGraphics(); g2.setColor(c); g2.fillRect(0, 0, 10, 10); return new ImageIcon(image); } private void createMenus(JComponent menu) { final JMenuItem addBam = new JMenuItem("Add BAM ..."); menu.add(addBam); addBam.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { FileSelectionDialog bamFileSelection = new FileSelectionDialog( null, false, "BamView", "BAM"); List<String> bamFiles = bamFileSelection.getFiles(BAM_SUFFIX); short count = (short) bamList.size(); bamList.addAll(bamFileSelection.getFiles(BAM_SUFFIX)); for(short i=0; i<bamFiles.size(); i++) addToViewMenu((short) (i+count)); laststart = -1; repaint(); } }); bamFilesMenu.setFont(addBam.getFont()); final JMenuItem groupBams = new JMenuItem("Group BAMs ..."); groupBams.addActionListener(new ActionListener(){ public void actionPerformed(ActionEvent arg0) { groupsFrame.updateAndDisplay(); } }); bamFilesMenu.add(groupBams); bamFilesMenu.addSeparator(); menu.add(bamFilesMenu); final JMenu analyse = new JMenu("Analyse"); menu.add(analyse); final JMenuItem readCount = new JMenuItem("Read count of selected features ..."); analyse.add(readCount); if(feature_display == null) readCount.setEnabled(false); readCount.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { FeatureVector features = feature_display.getSelection().getAllFeatures(); JCheckBox overlap = new JCheckBox("Include all overlapping reads", true); overlap.setToolTipText("Include reads that partially overlap the feature"); JCheckBox spliced = new JCheckBox("Introns included", true); Box yBox = Box.createVerticalBox(); yBox.add(overlap); yBox.add(spliced); final ReadCountDialog opts = new ReadCountDialog(new JFrame(), "Read Count Options", feature_display, yBox); if(opts.getStatus() == -1) return; //JOptionPane.showMessageDialog(null, yBox, "Read Count Option", JOptionPane.INFORMATION_MESSAGE); new MappedReads(features, (String)combo.getSelectedItem(), samFileReaderHash, bamList, seqNames, offsetLengths, concatSequences, seqLengths, samRecordFlagPredicate, samRecordMapQPredicate, !overlap.isSelected(), spliced.isSelected()); } }); final JMenuItem rpkm = new JMenuItem("RPKM value of selected features ..."); analyse.add(rpkm); if(feature_display == null) rpkm.setEnabled(false); rpkm.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { final FeatureVector features = feature_display.getSelection().getAllFeatures(); JCheckBox overlap = new JCheckBox("Include all overlapping reads", true); overlap.setToolTipText("Include reads that partially overlap the feature"); JCheckBox spliced = new JCheckBox("Introns included", true); JCheckBox allRefSeqs = new JCheckBox("Use reads mapped to all reference sequences", false); Box yBox = Box.createVerticalBox(); yBox.add(overlap); yBox.add(spliced); if(seqLengths.size() > 1) yBox.add(allRefSeqs); final ReadCountDialog opts = new ReadCountDialog(new JFrame(), "RPKM Options", feature_display, yBox); if(opts.getStatus() == -1) return; int seqlen = 0; if(feature_display != null) seqlen = feature_display.getSequenceLength(); else if(bases != null) seqlen = bases.getLength(); new MappedReads(features, (String)combo.getSelectedItem(), samFileReaderHash, bamList, seqNames, offsetLengths, concatSequences, seqLengths, seqlen, samRecordFlagPredicate, samRecordMapQPredicate, !overlap.isSelected(), spliced.isSelected(), allRefSeqs.isSelected()); } }); final JMenuItem createFeatures = new JMenuItem("Create features from coverage peaks ..."); analyse.add(createFeatures); if(feature_display == null) createFeatures.setEnabled(false); createFeatures.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if(feature_display == null) return; new CreateFeatures(groupsFrame); } }); for(short i=0; i<bamList.size(); i++) addToViewMenu(i); menu.add(new JSeparator()); JMenu viewMenu = new JMenu("Views"); cbStackView.setFont(viewMenu.getFont()); cbIsizeStackView.setFont(viewMenu.getFont()); cbPairedStackView.setFont(viewMenu.getFont()); cbStrandStackView.setFont(viewMenu.getFont()); cbCoverageView.setFont(viewMenu.getFont()); cbCoverageStrandView.setFont(viewMenu.getFont()); cbCoverageHeatMap.setFont(viewMenu.getFont()); baseQualityColour.setFont(viewMenu.getFont()); colourByCoverageColour.setFont(viewMenu.getFont()); markInsertions.setFont(viewMenu.getFont()); cbIsizeStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; logMenuItem.setEnabled(isIsizeStackView()); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED); repaint(); } }); viewMenu.add(cbIsizeStackView); cbStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(cbStackView.isSelected()) logMenuItem.setEnabled(false); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED); repaint(); } }); viewMenu.add(cbStackView); cbPairedStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(cbPairedStackView.isSelected()) logMenuItem.setEnabled(false); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED); repaint(); } }); viewMenu.add(cbPairedStackView); cbStrandStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(isStrandStackView()) setViewportMidPoint(); if(cbStrandStackView.isSelected()) logMenuItem.setEnabled(false); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED); repaint(); } }); viewMenu.add(cbStrandStackView); cbCoverageView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(cbCoverageView.isSelected()) { logMenuItem.setEnabled(true); coverageView.setPlotHeatMap(false); coverageView.setPlotByStrand(false); setViewportBtm(); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_NEVER); } repaint(); } }); viewMenu.add(cbCoverageView); cbCoverageStrandView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(cbCoverageStrandView.isSelected()) { logMenuItem.setEnabled(true); coverageView.setPlotHeatMap(false); coverageView.setPlotByStrand(true); setViewportBtm(); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_NEVER); } repaint(); } }); viewMenu.add(cbCoverageStrandView); cbCoverageHeatMap.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; if(cbCoverageHeatMap.isSelected()) { logMenuItem.setEnabled(true); coverageView.setPlotHeatMap(true); setViewportBtm(); getJspView().setVerticalScrollBarPolicy(JScrollPane.VERTICAL_SCROLLBAR_NEVER); } repaint(); } }); viewMenu.add(cbCoverageHeatMap); menu.add(viewMenu); final JCheckBoxMenuItem checkBoxSNPs = new JCheckBoxMenuItem("SNP marks"); // JMenu colourMenu = new JMenu("Colour By"); colourMenu.add(colourByCoverageColour); baseQualityColour.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if(baseQualityColour.isSelected()) { checkBoxSNPs.setSelected(false); isSNPs = false; } repaint(); } }); colourMenu.add(baseQualityColour); menu.add(colourMenu); // JMenu showMenu = new JMenu("Show"); JCheckBoxMenuItem checkBoxOrientation = new JCheckBoxMenuItem("Orientation"); checkBoxOrientation.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { isOrientation = !isOrientation; repaint(); } }); showMenu.add(checkBoxOrientation); JCheckBoxMenuItem checkBoxSingle = new JCheckBoxMenuItem("Single Reads"); checkBoxSingle.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { repaint(); isSingle = !isSingle; } }); showMenu.add(checkBoxSingle); checkBoxSNPs.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if (isSNPs && bases == null) { JOptionPane.showMessageDialog(null, "No reference sequence supplied to identify SNPs.", "SNPs", JOptionPane.INFORMATION_MESSAGE); } isSNPs = !isSNPs; if(isSNPs) baseQualityColour.setSelected(false); repaint(); } }); showMenu.add(checkBoxSNPs); markInsertions.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { repaint(); } }); showMenu.add(markInsertions); menu.add(showMenu); // JMenu graphMenu = new JMenu("Graph"); JCheckBoxMenuItem checkBoxCoverage = new JCheckBoxMenuItem("Coverage"); checkBoxCoverage.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { isCoverage = !isCoverage; coveragePanel.setVisible(isCoverage); if( isCoverage && !cbCoverageView.isSelected() && !cbCoverageStrandView.isSelected() && !cbCoverageHeatMap.isSelected()) laststart = -1; repaint(); } }); graphMenu.add(checkBoxCoverage); JCheckBoxMenuItem checkBoxSNP = new JCheckBoxMenuItem("SNP"); checkBoxSNP.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { isSNPplot = !isSNPplot; snpPanel.setVisible(isSNPplot); laststart = -1; repaint(); } }); graphMenu.add(checkBoxSNP); menu.add(graphMenu); if(feature_display != null) { final JCheckBoxMenuItem checkBoxSync = new JCheckBoxMenuItem("Asynchronous", asynchronous); checkBoxSync.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { asynchronous = checkBoxSync.isSelected(); } }); menu.add(checkBoxSync); } menu.add(new JSeparator()); JMenu maxHeightMenu = new JMenu("BamView Height"); menu.add(maxHeightMenu); final String hgts[] = {"500", "800", "1000", "1500", "2500", "5000", "50000"}; ButtonGroup bgroup = new ButtonGroup(); for(int i=0; i<hgts.length; i++) { final String hgt = hgts[i]; final JCheckBoxMenuItem maxHeightMenuItem = new JCheckBoxMenuItem(hgt); bgroup.add(maxHeightMenuItem); maxHeightMenuItem.setSelected(hgts[i].equals(Integer.toString(maxHeight))); maxHeightMenu.add(maxHeightMenuItem); maxHeightMenuItem.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if(maxHeightMenuItem.isSelected()) maxHeight = Integer.parseInt(hgt); int start = getBaseAtStartOfView(); setDisplay(start, nbasesInView+start, null); } }); } menu.add(new JSeparator()); logMenuItem.setFont(menu.getFont()); menu.add(logMenuItem); logMenuItem.setEnabled(isIsizeStackView()); logMenuItem.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { logScale = logMenuItem.isSelected(); repaint(); } }); JMenuItem filter = new JMenuItem("Filter Reads ..."); menu.add(filter); filter.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if(filterFrame == null) filterFrame = new SAMRecordFilter(BamView.this); else filterFrame.setVisible(true); } }); JMenuItem maxReadCoverage = new JMenuItem("Read Coverage Threshold ..."); menu.add(maxReadCoverage); maxReadCoverage.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { final TextFieldInt maxRead = new TextFieldInt(); maxRead.setValue(MAX_COVERAGE); int status = JOptionPane.showConfirmDialog(null, maxRead, "Read Coverage Threshold", JOptionPane.OK_CANCEL_OPTION); if(status == JOptionPane.OK_OPTION && maxRead.getValue() != MAX_COVERAGE) { MAX_COVERAGE = maxRead.getValue(); if(MAX_COVERAGE < 1) MAX_COVERAGE = Integer.MAX_VALUE; laststart = -1; repaint(); } } }); JMenuItem readList = new JMenuItem("List Reads ..."); menu.add(readList); readList.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { new SAMRecordList(BamView.this); } }); final JMenuItem bamSplitter = new JMenuItem("Clone window"); bamSplitter.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { BamView bamView = new BamView(new Vector<String>(bamList), null, nbasesInView, entry_edit, feature_display, bases, (JPanel) mainPanel.getParent(), null); bamView.getJspView().getVerticalScrollBar().setValue( bamView.getJspView().getVerticalScrollBar().getMaximum()); getJspView().getVerticalScrollBar().setValue( bamView.getJspView().getVerticalScrollBar().getMaximum()); int start = getBaseAtStartOfView(); setDisplay(start, nbasesInView+start, null); if(feature_display != null) { feature_display.addDisplayAdjustmentListener(bamView); feature_display.getSelection().addSelectionChangeListener(bamView); } } }); menu.add(new JSeparator()); menu.add(bamSplitter); // JMenu coverageMenu = new JMenu("Coverage Options"); coverageView.init(this, 0.f, 0, 0); coverageView.createMenus(coverageMenu); viewMenu.add(new JSeparator()); viewMenu.add(coverageMenu); } private JComponent bamTopPanel(final JFrame frame) { final JComponent topPanel; if(frame == null) { topPanel = new JPanel(new FlowLayout(FlowLayout.LEADING, 0, 0)); if(feature_display != null) this.selection = feature_display.getSelection(); } else { topPanel = new JMenuBar(); frame.setJMenuBar((JMenuBar)topPanel); JMenu fileMenu = new JMenu("File"); topPanel.add(fileMenu); JMenuItem readBam = new JMenuItem("Open new BamView ..."); fileMenu.add(readBam); readBam.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { String[] s = { "NEW-BAMVIEW" }; BamView.main(s); } }); JMenuItem saveAs = new JMenuItem("Save As Image File (png/jpeg/svg) ..."); fileMenu.add(saveAs); saveAs.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { PrintBamView.print((JPanel)mainPanel.getParent()); } }); JMenuItem close = new JMenuItem("Close"); fileMenu.add(close); close.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { BamView.this.setVisible(false); Component comp = BamView.this; while( !(comp instanceof JFrame) ) comp = comp.getParent(); ((JFrame)comp).dispose(); } }); JMenuItem exit = new JMenuItem("Exit"); fileMenu.add(new JSeparator()); fileMenu.add(exit); exit.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { int status = JOptionPane.showConfirmDialog(BamView.this, "Exit BamView?", "Exit", JOptionPane.OK_CANCEL_OPTION); if(status != JOptionPane.OK_OPTION) return; System.exit(0); } }); addKeyListener(new KeyAdapter() { public void keyPressed(final KeyEvent event) { switch (event.getKeyCode()) { case KeyEvent.VK_UP: setZoomLevel((int) (BamView.this.nbasesInView * 1.1)); break; case KeyEvent.VK_DOWN: if (showBaseAlignment) break; setZoomLevel((int) (BamView.this.nbasesInView * .9)); break; default: break; } } }); } if(seqNames.size() > 1) { int len = 0; for(int i=0; i<seqNames.size(); i++) len += seqLengths.get(seqNames.get(i)); if(feature_display != null && len == feature_display.getSequenceLength()) concatSequences = true; else if(bases != null && len == bases.getLength() ) concatSequences = true; } // auto hide top panel final JCheckBox buttonAutoHide = new JCheckBox("Hide", (frame == null)); buttonAutoHide.setToolTipText("Auto-Hide"); final MouseMotionListener mouseMotionListener = new MouseMotionListener() { public void mouseDragged(MouseEvent event) { handleCanvasMouseDrag(event); } public void mouseMoved(MouseEvent e) { lastMousePoint = e.getPoint(); int thisHgt = HEIGHT; if (thisHgt < 5) thisHgt = 15; int y = (int) (e.getY() - jspView.getViewport().getViewRect().getY()); if (y < thisHgt) { topPanel.setVisible(true); } else { if (buttonAutoHide.isSelected()) topPanel.setVisible(false); } mainPanel.repaint(); mainPanel.revalidate(); } }; addMouseMotionListener(mouseMotionListener); combo = new SequenceComboBox(seqNames){ private static final long serialVersionUID = 1L; public void update(IndexReferenceEvent event) { laststart = -1; if(feature_display != null) setZoomLevel(feature_display.getMaxVisibleBases()); else setZoomLevel(BamView.this.nbasesInView); } }; topPanel.add(combo); if(feature_display == null) { final JTextField baseText = new JTextField(8); JButton goTo = new JButton("GoTo:"); goTo.setToolTipText("Go to base position"); goTo.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { try { int basePosition = Integer.parseInt(baseText.getText()); scrollBar.setValue(basePosition); } catch (NumberFormatException nfe) { JOptionPane.showMessageDialog(BamView.this, "Expecting a base number!", "Number Format", JOptionPane.WARNING_MESSAGE); } } }); topPanel.add(goTo); topPanel.add(baseText); JButton zoomIn = new JButton("-"); zoomIn.setToolTipText("Zoom out (up arrow)"); Insets ins = new Insets(1,1,1,1); zoomIn.setMargin(ins); zoomIn.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { setZoomLevel((int) (BamView.this.nbasesInView * 1.1)); } }); topPanel.add(zoomIn); JButton zoomOut = new JButton("+"); zoomOut.setToolTipText("Zoom in (down arrow)"); zoomOut.setMargin(ins); zoomOut.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if (showBaseAlignment) return; setZoomLevel((int) (BamView.this.nbasesInView * .9)); } }); topPanel.add(zoomOut); } topPanel.add(buttonAutoHide); if(feature_display != null) { JButton close = new JButton("Close"); topPanel.add(close); close.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { final JPanel containerPanel = (JPanel) mainPanel.getParent(); feature_display.removeDisplayAdjustmentListener(BamView.this); feature_display.getSelection().removeSelectionChangeListener(BamView.this); containerPanel.remove(mainPanel); if(containerPanel.getComponentCount() > 0) containerPanel.revalidate(); else { if(entry_edit != null) entry_edit.setNGDivider(); else containerPanel.setVisible(false); } } }); } return topPanel; } public void setVisible(boolean visible) { super.setVisible(visible); mainPanel.setVisible(visible); } private void setViewportMidPoint() { Point p = jspView.getViewport().getLocation(); p.y = (getHeight() - jspView.getViewport().getViewRect().height)/2; jspView.getViewport().setViewPosition(p); } private void setViewportBtm() { jspView.getVerticalScrollBar().setValue( jspView.getVerticalScrollBar().getMaximum()); } protected int getBaseAtStartOfView() { if(feature_display != null) return feature_display.getForwardBaseAtLeftEdge(); else return scrollBar.getValue(); } /** * Set the panel size based on the number of bases visible * and repaint. * @param nbasesInView */ private void setZoomLevel(final int nbasesInView) { int startValue = getBaseAtStartOfView(); this.nbasesInView = nbasesInView; float pixPerBase = getPixPerBaseByWidth(); if(isBaseAlignmentView(pixPerBase)) { pixPerBase = ALIGNMENT_PIX_PER_BASE; this.nbasesInView = (int)(mainPanel.getWidth()/pixPerBase); jspView.getVerticalScrollBar().setValue(0); if(ruler == null) ruler = new Ruler(); jspView.setColumnHeaderView(ruler); showBaseAlignment = true; baseQualityColour.setEnabled(true); markInsertions.setEnabled(true); } else if(jspView != null) { if(isCoverageView(pixPerBase) && nbasesInView >= MAX_BASES) { cbLastSelected = getSelectedCheckBoxMenuItem(); cbCoverageView.setSelected(true); coverageView.setPlotByStrand(false); } else if(isCoverageView(pixPerBase) && nbasesInView < MAX_BASES && cbLastSelected != null) { cbLastSelected.setSelected(true); cbLastSelected = null; } jspView.setColumnHeaderView(null); if(!isStrandStackView()) setViewportBtm(); else setViewportMidPoint(); showBaseAlignment = false; baseQualityColour.setEnabled(false); markInsertions.setEnabled(false); } if(scrollBar != null) { scrollBar.setValues(startValue, nbasesInView, 1, getMaxBasesInPanel(getSequenceLength())); scrollBar.setUnitIncrement(nbasesInView/20); scrollBar.setBlockIncrement(nbasesInView); } } /** * Set the start and end base positions to display. * @param start * @param end * @param event */ public void setDisplay(int start, int end, DisplayAdjustmentEvent event) { this.startBase = start; this.endBase = end; this.nbasesInView = end-start+1; lastMousePoint = null; float pixPerBase; if(jspView.getViewport().getViewRect().width > 0) pixPerBase = getPixPerBaseByWidth(); else { if(feature_display == null) pixPerBase = 1000.f/(float)(end-start+1); else pixPerBase = feature_display.getWidth()/(float)(end-start+1); } Dimension d = new Dimension(); d.setSize(nbasesInView*pixPerBase, maxHeight); setPreferredSize(d); if(event == null) { this.startBase = -1; this.endBase = -1; } } /** * Return an Artemis entry from a file * @param entryFileName * @param entryGroup * @return * @throws NoSequenceException */ private Entry getEntry(final String entryFileName, final EntryGroup entryGroup) throws NoSequenceException { final Document entry_document = DocumentFactory.makeDocument(entryFileName); final EntryInformation artemis_entry_information = Options.getArtemisEntryInformation(); //System.out.println(entryFileName); final uk.ac.sanger.artemis.io.Entry new_embl_entry = EntryFileDialog.getEntryFromFile(null, entry_document, artemis_entry_information, false); if(new_embl_entry == null) // the read failed return null; Entry entry = null; try { if(entryGroup.getSequenceEntry() != null) bases = entryGroup.getSequenceEntry().getBases(); if(bases == null) { entry = new Entry(new_embl_entry); bases = entry.getBases(); } else entry = new Entry(bases,new_embl_entry); entryGroup.add(entry); } catch(OutOfRangeException e) { new MessageDialog(null, "read failed: one of the features in " + entryFileName + " has an out of range " + "location: " + e.getMessage()); } return entry; } private boolean isShowScale() { return (feature_display == null ? true : false); } public JScrollPane getJspView() { return jspView; } /** * Handle a mouse drag event on the drawing canvas. **/ private void handleCanvasMouseDrag(final MouseEvent event) { if(event.getButton() == MouseEvent.BUTTON3 || bases == null) return; highlightSAMRecord = null; if(event.getClickCount() > 1) { getSelection().clear(); repaint(); return; } highlightRange(event, MouseEvent.BUTTON1_DOWN_MASK & MouseEvent.BUTTON2_DOWN_MASK); } /** * * @param event * @param onmask */ protected void highlightRange(MouseEvent event, int onmask) { int seqLength = getSequenceLength(); float pixPerBase = getPixPerBaseByWidth(); int start = (int) ( ( (event.getPoint().getX())/pixPerBase) + getBaseAtStartOfView() ); if(start < 1) start = 1; if(start > seqLength) start = seqLength; if (dragStart < 0 && (event.getModifiersEx() & onmask) == onmask) dragStart = start; else if((event.getModifiersEx() & onmask) != onmask) dragStart = -1; MarkerRange drag_range; try { if(dragStart < 0) drag_range = new MarkerRange (bases.getForwardStrand(), start, start); else drag_range = new MarkerRange (bases.getForwardStrand(), dragStart, start); getSelection().setMarkerRange(drag_range); repaint(); } catch (OutOfRangeException e) { e.printStackTrace(); } } /** * Get the colour for the given read given to it by the coverage plot. * @param samRecord * @return */ private Color getColourByCoverageColour(BamViewRecord samRecord) { short fileIndex = 0; if(bamList.size()>1) fileIndex = samRecord.bamIndex; return getColourByCoverageColour(fileIndex); } private Color getColourByCoverageColour(final short fileIndex) { LineAttributes lines[] = CoveragePanel.getLineAttributes(bamList.size()); return lines[fileIndex].getLineColour(); } protected int getMaxBases() { return MAX_BASES; } protected void setMaxBases(int max) { MAX_BASES = max; } private boolean isStackView() { return cbStackView.isSelected(); } private boolean isPairedStackView() { return cbPairedStackView.isSelected(); } private boolean isStrandStackView() { return cbStrandStackView.isSelected(); } private boolean isCoverageView(float pixPerBase) { if(isBaseAlignmentView(pixPerBase)) return false; return cbCoverageView.isSelected() || cbCoverageStrandView.isSelected() || cbCoverageHeatMap.isSelected(); } private boolean isIsizeStackView() { return cbIsizeStackView.isSelected(); } private boolean isBaseAlignmentView(float pixPerBase) { if(pixPerBase*1.08f >= ALIGNMENT_PIX_PER_BASE) return true; return false; } private JCheckBoxMenuItem getSelectedCheckBoxMenuItem() { if(isStackView()) return cbStackView; if(isPairedStackView()) return cbPairedStackView; if(isStrandStackView()) return cbStrandStackView; if(isIsizeStackView()) return cbIsizeStackView; if(cbCoverageView.isSelected()) return cbCoverageView; if(cbCoverageHeatMap.isSelected()) return cbCoverageHeatMap; return cbCoverageStrandView; } protected Selection getSelection() { return selection; } protected List<BamViewRecord> getReadsInView() { return readsInView; } protected int getBasesInView() { return nbasesInView; } protected void setHighlightSAMRecord(BamViewRecord highlightSAMRecord) { this.highlightSAMRecord = highlightSAMRecord; } protected BamViewRecord getHighlightSAMRecord() { return highlightSAMRecord; } protected FeatureDisplay getFeatureDisplay() { return feature_display; } /** * @return the combo */ public SequenceComboBox getCombo() { return combo; } private String getVersion() { final ClassLoader cl = this.getClass().getClassLoader(); try { String line; InputStream in = cl.getResourceAsStream("etc/versions"); BufferedReader reader = new BufferedReader(new InputStreamReader(in)); while((line = reader.readLine()) != null) { if(line.startsWith("BamView")) return line.substring( "BamView".length() ).trim(); } reader.close(); in.close(); } catch (Exception ex) { } return null; } /** * Artemis event notification */ public void displayAdjustmentValueChanged(final DisplayAdjustmentEvent event) { if(event.getType() == DisplayAdjustmentEvent.REV_COMP_EVENT && event.isRevCompDisplay()) JOptionPane.showMessageDialog(this, "Flipping the display is not supported by BamView.", "Warning", JOptionPane.WARNING_MESSAGE); if(!asynchronous) { // if not asynchronous displayAdjustmentWork(event); return; } SwingWorker worker = new SwingWorker() { public Object construct() { try { Thread.sleep(500); } catch (InterruptedException e) { e.printStackTrace(); } if(event.getStart() != ((FeatureDisplay)event.getSource()).getForwardBaseAtLeftEdge()) { waitingFrame.showWaiting("waiting...", mainPanel); return null; } displayAdjustmentWork(event); waitingFrame.setVisible(false); return null; } }; worker.start(); } /** * Carry out the display agjustment event action. * @param event */ private void displayAdjustmentWork(final DisplayAdjustmentEvent event) { if(event.getType() == DisplayAdjustmentEvent.SCALE_ADJUST_EVENT) { laststart = -1; BamView.this.startBase = event.getStart(); BamView.this.endBase = event.getEnd(); int width = feature_display.getMaxVisibleBases(); setZoomLevel(width); repaint(); } else { setDisplay(event.getStart(), event.getStart()+feature_display.getMaxVisibleBases(), event); repaint(); } } public void selectionChanged(SelectionChangeEvent event) { repaint(); } private class Ruler extends JPanel { private static final long serialVersionUID = 1L; protected int start; protected int end; protected String refSeq; public Ruler() { super(); setPreferredSize(new Dimension(mainPanel.getWidth(), 26)); setBackground(Color.white); } public void paintComponent(Graphics g) { super.paintComponent(g); Graphics2D g2 = (Graphics2D)g; drawBaseScale(g2, start, end, 12); } private void drawBaseScale(Graphics2D g2, int start, int end, int ypos) { int startMark = (((int)(start/10))*10)+1; if(end > getSequenceLength()) end = getSequenceLength(); for(int i=startMark; i<end; i+=20) { int xpos = (i-start)*ALIGNMENT_PIX_PER_BASE; g2.drawString(Integer.toString(i), xpos, ypos); } for(int i=startMark; i<end; i+=10) { int xpos = (i-start)*ALIGNMENT_PIX_PER_BASE; xpos+=(ALIGNMENT_PIX_PER_BASE/2); g2.drawLine(xpos, ypos+1, xpos, ypos+5); } if(refSeq != null) { ypos+=15; g2.setColor(LIGHT_GREY); g2.fillRect(0, ypos-11, getWidth(), 11); g2.translate(0, 16); drawSelectionRange(g2, ALIGNMENT_PIX_PER_BASE, start, end, Color.yellow); g2.translate(0, -16); g2.setColor(Color.black); g2.drawString(refSeq, 0, ypos-2); } } } /** * Popup menu listener */ class PopupListener extends MouseAdapter { private JMenuItem gotoMateMenuItem; private JMenuItem showDetails; private JMenu coverageMenu; private JMenuItem createGroup; public void mouseClicked(MouseEvent e) { if(e.isPopupTrigger() || e.getButton() == MouseEvent.BUTTON3) return; BamView.this.requestFocus(); if(e.getClickCount() > 1) getSelection().clear(); else if(e.getButton() == MouseEvent.BUTTON1) { if(isCoverageView(getPixPerBaseByWidth())) coverageView.singleClick(e.isShiftDown(), e.getPoint().y-getJspView().getViewport().getViewPosition().y); else highlightSAMRecord = mouseOverSAMRecord; } else highlightRange(e, MouseEvent.BUTTON2_DOWN_MASK); repaint(); } public void mousePressed(MouseEvent e) { maybeShowPopup(e); } public void mouseReleased(MouseEvent e) { dragStart = -1; maybeShowPopup(e); } private void maybeShowPopup(MouseEvent e) { if(e.isPopupTrigger()) { // // main menu options if(popup == null) { popup = new JPopupMenu(); createMenus(popup); } // // coverage heatmap menu options if(coverageMenu != null) popup.remove(coverageMenu); if(isCoverageView(getPixPerBaseByWidth()) && coverageView.isPlotHeatMap()) { if(coverageMenu == null) { coverageMenu = new JMenu("Coverage HeatMap"); coverageView.createMenus(coverageMenu); final JCheckBoxMenuItem coverageGrid = new JCheckBoxMenuItem("Show heatmap grid", false); coverageGrid.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { coverageView.showLabels(coverageGrid.isSelected()); } }); coverageMenu.add(coverageGrid); createGroup = new JMenuItem("Create group from selected BAMs"); createGroup.addActionListener(new ActionListener() { private int n = 1; public void actionPerformed(ActionEvent e) { String groupName = "group_"+n; groupsFrame.addGroup(groupName); final List<String> selected = coverageView.getSelected(); for(String sel: selected) groupsFrame.addToGroup((new File(sel)).getName(), groupName); groupsFrame.updateAndDisplay(); n++; } }); coverageMenu.add(createGroup); } createGroup.setEnabled(coverageView.hasSelectedBams()); popup.add(coverageMenu); } if(gotoMateMenuItem != null) popup.remove(gotoMateMenuItem); if(showDetails != null) popup.remove(showDetails); if( mouseOverSAMRecord != null && mouseOverSAMRecord.sam.getReadPairedFlag() && !mouseOverSAMRecord.sam.getMateUnmappedFlag() ) { final BamViewRecord thisSAMRecord = mouseOverSAMRecord; gotoMateMenuItem = new JMenuItem("Go to mate of : "+ thisSAMRecord.sam.getReadName()); gotoMateMenuItem.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { String name = thisSAMRecord.sam.getMateReferenceName(); if(name.equals("=")) name = thisSAMRecord.sam.getReferenceName(); int offset = getSequenceOffset(name); if(feature_display != null) feature_display.makeBaseVisible( thisSAMRecord.sam.getMateAlignmentStart()+offset); else scrollBar.setValue( thisSAMRecord.sam.getMateAlignmentStart()+offset- (nbasesInView/2)); highlightSAMRecord = thisSAMRecord; } }); popup.add(gotoMateMenuItem); } if( mouseOverSAMRecord != null) { final BamViewRecord thisSAMRecord = mouseOverSAMRecord; showDetails = new JMenuItem("Show details of : "+ thisSAMRecord.sam.getReadName()); showDetails.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { openFileViewer(thisSAMRecord.sam, getMate(thisSAMRecord), bamList); } }); popup.add(showDetails); } popup.show(e.getComponent(), e.getX(), e.getY()); } } } protected static void openFileViewer(SAMRecord readRecord, SAMRecord mateRecord, List<String> bamList) { FileViewer viewDetail = new FileViewer(readRecord.getReadName(), true, false, false); appendToDetailView(readRecord, mateRecord, viewDetail, bamList); } private static void appendToDetailView(final SAMRecord thisSAMRecord, final SAMRecord thisSAMRecordMate, final FileViewer viewDetail, final List<String> bamList) { if(bamList.size() > 1 && thisSAMRecord.getAttribute("FL") != null) { int bamIndex = (Integer)thisSAMRecord.getAttribute("FL"); if(bamIndex < bamList.size()) viewDetail.appendString("File "+bamList.get(bamIndex)+"\n\n", Level.INFO); } viewDetail.appendString("Read Name "+thisSAMRecord.getReadName()+"\n", Level.INFO); viewDetail.appendString("Coordinates "+thisSAMRecord.getAlignmentStart()+".."+ thisSAMRecord.getAlignmentEnd()+"\n", Level.DEBUG); viewDetail.appendString("Length "+thisSAMRecord.getReadLength()+"\n", Level.DEBUG); viewDetail.appendString("Reference Name "+thisSAMRecord.getReferenceName()+"\n", Level.DEBUG); viewDetail.appendString("Inferred Size "+thisSAMRecord.getInferredInsertSize()+"\n", Level.DEBUG); viewDetail.appendString("Mapping Quality "+thisSAMRecord.getMappingQuality()+"\n", Level.DEBUG); viewDetail.appendString("Cigar String "+thisSAMRecord.getCigarString()+"\n", Level.DEBUG); viewDetail.appendString("Strand "+ (thisSAMRecord.getReadNegativeStrandFlag() ? "-\n\n" : "+\n\n"), Level.DEBUG); if(thisSAMRecord.getReadPairedFlag() && !thisSAMRecord.getMateUnmappedFlag()) { if(thisSAMRecordMate != null) { viewDetail.appendString("Mate Coordinates "+thisSAMRecordMate.getAlignmentStart()+".."+ thisSAMRecordMate.getAlignmentEnd()+"\n", Level.DEBUG); viewDetail.appendString("Mate Length "+thisSAMRecordMate.getReadLength()+"\n", Level.DEBUG); viewDetail.appendString("Mate Reference Name "+thisSAMRecordMate.getReferenceName()+"\n", Level.DEBUG); viewDetail.appendString("Mate Inferred Size "+thisSAMRecordMate.getInferredInsertSize()+"\n", Level.DEBUG); viewDetail.appendString("Mate Mapping Quality "+thisSAMRecordMate.getMappingQuality()+"\n", Level.DEBUG); viewDetail.appendString("Mate Cigar String "+thisSAMRecordMate.getCigarString()+"\n", Level.DEBUG); } else { viewDetail.appendString("Mate Start Coordinate "+thisSAMRecord.getMateAlignmentStart()+"\n", Level.DEBUG); viewDetail.appendString("Mate Reference Name "+thisSAMRecord.getMateReferenceName()+"\n", Level.DEBUG); } viewDetail.appendString("Mate Strand "+ (thisSAMRecord.getMateNegativeStrandFlag() ? "-" : "+"), Level.DEBUG); } else { viewDetail.appendString("Mate Unmapped ", Level.DEBUG); } viewDetail.appendString("\n\nFlags:", Level.INFO); viewDetail.appendString("\nDuplicate Read "+ (thisSAMRecord.getDuplicateReadFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\nRead Paired "+ (thisSAMRecord.getReadPairedFlag() ? "yes" : "no"), Level.DEBUG); if(thisSAMRecord.getReadPairedFlag()) { viewDetail.appendString("\nFirst of Pair "+ (thisSAMRecord.getFirstOfPairFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\nMate Unmapped "+ (thisSAMRecord.getMateUnmappedFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\nProper Pair "+ (thisSAMRecord.getProperPairFlag() ? "yes" : "no"), Level.DEBUG); } viewDetail.appendString("\nRead Fails Vendor\nQuality Check "+ (thisSAMRecord.getReadFailsVendorQualityCheckFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\nRead Unmapped "+ (thisSAMRecord.getReadUnmappedFlag() ? "yes" : "no"), Level.DEBUG); if(thisSAMRecord.getReadPairedFlag()) viewDetail.appendString("\nSecond Of Pair "+ (thisSAMRecord.getSecondOfPairFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\n\nRead Bases:\n", Level.INFO); // wrap the read bases String seq = new String(thisSAMRecord.getReadBases()); int MAX_SEQ_LINE_LENGTH = 100; for(int i=0; i<=seq.length(); i+=MAX_SEQ_LINE_LENGTH) { int iend = i+MAX_SEQ_LINE_LENGTH; if(iend > seq.length()) iend = seq.length(); viewDetail.appendString(seq.substring(i, iend)+"\n", Level.DEBUG); } } /** * Query for the mate of a read * @param mate * @return */ protected SAMRecord getMate(BamViewRecord thisSAMRecord) { if(!thisSAMRecord.sam.getReadPairedFlag() || // read is not paired in sequencing thisSAMRecord.sam.getMateUnmappedFlag()) return null; SAMRecord mate = null; try { short fileIndex = 0; if(bamList.size()>1 && thisSAMRecord.bamIndex > 0) fileIndex = thisSAMRecord.bamIndex; String bam = bamList.get(fileIndex); final SAMFileReader inputSam = getSAMFileReader(bam); mate = inputSam.queryMate(thisSAMRecord.sam); } catch (Exception e) { e.printStackTrace(); } return mate; } protected SAMRecordPredicate getSamRecordFlagPredicate() { return samRecordFlagPredicate; } protected void setSamRecordFlagPredicate( SAMRecordPredicate samRecordFlagPredicate) { laststart = -1; lastend = -1; this.samRecordFlagPredicate = samRecordFlagPredicate; } protected SAMRecordMapQPredicate getSamRecordMapQPredicate() { return samRecordMapQPredicate; } protected void setSamRecordMapQPredicate( SAMRecordMapQPredicate samRecordMapQPredicate) { laststart = -1; lastend = -1; this.samRecordMapQPredicate = samRecordMapQPredicate; } /** * @return the concatSequences */ protected boolean isConcatSequences() { return concatSequences; } class PairedRead { BamViewRecord sam1; BamViewRecord sam2; } class CreateFeatures { CreateFeatures(final GroupBamFrame groupsFrame) { final TextFieldInt threshold = new TextFieldInt(); final TextFieldInt minSize = new TextFieldInt(); final TextFieldInt minBams = new TextFieldInt(); threshold.setValue(6); minSize.setValue(6); minBams.setValue( (groupsFrame.getNumberOfGroups() == 1 ? bamList.size() : groupsFrame.getMaximumBamsInGroup()) ); final JPanel gridPanel = new JPanel(new GridBagLayout()); GridBagConstraints c = new GridBagConstraints(); c.anchor = GridBagConstraints.WEST; c.fill = GridBagConstraints.HORIZONTAL; c.gridx = 0; c.gridy = 0; gridPanel.add(new JLabel("Minimum number of reads:"), c); c.gridy++; gridPanel.add(threshold, c); c.gridy++; gridPanel.add(new JSeparator(), c); c.gridy++; gridPanel.add(new JLabel("Minimum number of BAMs for reads to be present in:"), c); c.gridy++; gridPanel.add(minBams, c); JRadioButton useAllBams = new JRadioButton("out of all BAMs", (groupsFrame.getNumberOfGroups() == 1)); JRadioButton useGroup = new JRadioButton("within a group", (groupsFrame.getNumberOfGroups() != 1)); if(groupsFrame.getNumberOfGroups() == 1) useGroup.setEnabled(false); final ButtonGroup group = new ButtonGroup(); group.add(useAllBams); group.add(useGroup); final Box xBox = Box.createHorizontalBox(); xBox.add(useAllBams); xBox.add(useGroup); xBox.add(Box.createHorizontalGlue()); c.gridy++; gridPanel.add(xBox, c); c.gridy++; gridPanel.add(new JSeparator(), c); c.gridy++; gridPanel.add(new JLabel("Minimum feature size:"), c); c.gridy++; gridPanel.add(minSize, c); final JCheckBox cbOpposite = new JCheckBox("Assume reads on opposite strand", false); cbOpposite.setToolTipText("for cDNA experiments when the reads are on the opposite strand"); c.gridy++; gridPanel.add(cbOpposite, c); int status = JOptionPane.showConfirmDialog(feature_display, gridPanel, "Options", JOptionPane.OK_CANCEL_OPTION); if(status == JOptionPane.CANCEL_OPTION) return; if(!useGroup.isSelected() && minBams.getValue() > bamList.size()) { status = JOptionPane.showConfirmDialog(feature_display, "The minimum number of BAMs setting can not be\n"+ "greater than the total number of BAM files.\n"+ "Set this to the number of BAMs (i.e. "+bamList.size()+").", "Options", JOptionPane.OK_CANCEL_OPTION); if(status == JOptionPane.CANCEL_OPTION) return; minBams.setValue(bamList.size()); } else if(useGroup.isSelected() && minBams.getValue() > groupsFrame.getMaximumBamsInGroup()) { status = JOptionPane.showConfirmDialog(feature_display, "Minimum number of BAMs setting can not be greater than\n"+ "the total number of BAM files found in any of the groups.\n"+ "Set this to the greatest number of BAM files in any\n"+ "group (i.e. "+groupsFrame.getMaximumBamsInGroup()+").", "Options", JOptionPane.OK_CANCEL_OPTION); if(status == JOptionPane.CANCEL_OPTION) return; minBams.setValue(groupsFrame.getMaximumBamsInGroup()); } new MappedReads((String)combo.getSelectedItem(),BamView.this, samFileReaderHash, seqNames, offsetLengths, concatSequences, seqLengths, (useGroup.isSelected() ? groupsFrame : null), threshold.getValue(), minSize.getValue(), minBams.getValue(), cbOpposite.isSelected(), true); } } public static void main(String[] args) { BamFrame frame = new BamFrame(); if(args.length == 0 && BamFrame.isMac()) { try { Thread.sleep(1000); } catch (InterruptedException e1) {} if(frame.getBamFile() != null) args = new String[]{ frame.getBamFile() }; } List<String> bam = new Vector<String>(); String reference = null; if(args.length == 0 || args[0].equals("NEW-BAMVIEW")) { System.setProperty("default_directory", System.getProperty("user.dir")); FileSelectionDialog fileSelection = new FileSelectionDialog( null, true, "BamView", "BAM"); bam = fileSelection.getFiles(BAM_SUFFIX); reference = fileSelection.getReferenceFile(); if(reference == null || reference.equals("")) reference = null; if(bam == null || bam.size() < 1) { if(args.length > 0 && args[0].equals("NEW-BAMVIEW")) return; System.err.println("No files found."); System.exit(0); } } else if(!args[0].startsWith("-")) { for(int i=0; i< args.length; i++) bam.add(args[i]); } int nbasesInView = 2000; String chr = null; String vw = null; boolean orientation = false; boolean covPlot = false; boolean snpPlot = false; int base = 0; for(int i=0;i<args.length; i++) { if(args[i].equals("-a")) { while(i < args.length-1 && !args[++i].startsWith("-")) { String filename = args[i]; if(FileSelectionDialog.isListOfFiles(filename)) bam.addAll(FileSelectionDialog.getListOfFiles(filename)); else bam.add(filename); } --i; } else if(args[i].equals("-r")) reference = args[++i]; else if(args[i].equals("-n")) nbasesInView = Integer.parseInt(args[++i]); else if(args[i].equals("-s")) System.setProperty("samtoolDir", args[++i]); else if(args[i].equals("-c")) chr = args[++i].trim(); else if(args[i].equals("-b")) base = Integer.parseInt(args[++i].trim()); else if(args[i].equals("-v")) vw = args[++i].trim(); else if(args[i].equals("-o")) orientation = true; else if(args[i].equals("-pc")) covPlot = true; else if(args[i].equals("-ps")) snpPlot = true; else if(args[i].startsWith("-h")) { System.out.println("-h\t show help"); System.out.println("-a\t BAM/SAM file to display"); System.out.println("-r\t reference file (optional)"); System.out.println("-n\t number of bases to display in the view (optional)"); System.out.println("-c\t chromosome name (optional)"); System.out.println("-v\t view (optional - IS (inferred size), S (stack, default), PS (paired stack), ST (strand), C (coverage))"); System.out.println("-b\t base position (optional)"); System.out.println("-o\t show orientation (optional)"); System.out.println("-pc\t plot coverage (optional)"); System.out.println("-ps\t plot SNP (optional and only with -r)"); System.exit(0); } } final BamView view = new BamView(bam, reference, nbasesInView, null, null, (JPanel)frame.getContentPane(), frame); frame.setTitle("BamView v"+view.getVersion()); if(chr != null) view.combo.setSelectedItem(chr); if(vw != null) { if(vw.equalsIgnoreCase("IS")) view.cbIsizeStackView.setSelected(true); if(vw.equalsIgnoreCase("PS")) view.cbPairedStackView.setSelected(true); if(vw.equalsIgnoreCase("ST")) view.cbStrandStackView.setSelected(true); if(vw.equalsIgnoreCase("C")) view.cbCoverageView.setSelected(true); } if(base > 0) view.scrollBar.setValue(base); if(orientation) view.isOrientation = true; if(covPlot) { view.isCoverage = true; view.coveragePanel.setVisible(true); } if(snpPlot) { view.isSNPplot = true; view.snpPanel.setVisible(true); } // translucent //frame.getRootPane().putClientProperty("Window.alpha", new Float(0.9f)); /*frame.addWindowFocusListener(new WindowFocusListener() { public void windowGainedFocus(WindowEvent e) { view.requestFocus(); } public void windowLostFocus(WindowEvent e){} });*/ frame.pack(); view.jspView.getVerticalScrollBar().setValue( view.jspView.getVerticalScrollBar().getMaximum()); frame.setVisible(true); } }