/* JamView * * 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.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.event.WindowEvent; import java.awt.event.WindowFocusListener; import java.io.File; import java.lang.management.ManagementFactory; import java.lang.management.MemoryMXBean; import java.util.Collections; import java.util.Enumeration; import java.util.Hashtable; import java.util.List; import java.util.Vector; import javax.swing.ButtonGroup; import javax.swing.JButton; import javax.swing.JCheckBox; import javax.swing.JCheckBoxMenuItem; import javax.swing.JComboBox; import javax.swing.JComponent; import javax.swing.JFrame; 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.JScrollBar; import javax.swing.JScrollPane; import javax.swing.JSeparator; import javax.swing.JTextField; import javax.swing.UIManager; import javax.swing.border.Border; import javax.swing.border.EmptyBorder; import org.apache.log4j.Level; import net.sf.samtools.AlignmentBlock; 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.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.components.DisplayAdjustmentEvent; import uk.ac.sanger.artemis.components.DisplayAdjustmentListener; 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.MessageDialog; 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.OutOfRangeException; public class BamView extends JPanel implements DisplayAdjustmentListener, SelectionChangeListener { private static final long serialVersionUID = 1L; private List<SAMRecord> readsInView; private Hashtable<String, Integer> seqLengths = new Hashtable<String, Integer>(); private Hashtable<String, Integer> offsetLengths; private Vector<String> seqNames = new Vector<String>(); private String bam; private SAMRecordFlagPredicate samRecordFlagPredicate; private SAMRecordMapQPredicate samRecordMapQPredicate; private Bases bases; private JScrollPane jspView; private JScrollBar scrollBar; private JComboBox combo; private boolean isSingle = false; private boolean isSNPs = false; private boolean isStackView = false; private boolean isPairedStackView = false; private boolean isStrandStackView = false; private boolean isCoverage = false; private FeatureDisplay feature_display; private Selection selection; private JPanel mainPanel; private CoveragePanel coveragePanel; private boolean showScale = true; private boolean logScale = false; private Ruler ruler; private int nbasesInView; private int startBase = -1; private int endBase = -1; private int laststart; private int lastend; private int maxUnitIncrement = 8; private boolean asynchronous = true; private boolean showBaseAlignment = false; private JCheckBoxMenuItem logMenuItem = new JCheckBoxMenuItem("Use Log Scale", logScale); private JCheckBoxMenuItem checkBoxStackView = new JCheckBoxMenuItem("Stack View"); private JCheckBoxMenuItem baseQualityColour = new JCheckBoxMenuItem("Colour by Base Quality");; private JCheckBoxMenuItem markInsertions = new JCheckBoxMenuItem("Mark Insertions"); private AlphaComposite translucent = AlphaComposite.getInstance(AlphaComposite.SRC_OVER, 0.6f); /** Used to colour the frames. */ private Color lightGrey = new Color(200, 200, 200); private Color darkGreen = new Color(0, 150, 0); private Color darkOrange = new Color(255,140,0); private Color deepPink = new Color(139,10,80); private Point lastMousePoint = null; private SAMRecord mouseOverSAMRecord = null; private SAMRecord highlightSAMRecord = null; private String mouseOverInsertion; // record of where a mouse drag starts private int dragStart = -1; 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..."); public static org.apache.log4j.Logger logger4j = org.apache.log4j.Logger.getLogger(BamView.class); public BamView(String bam, String reference, int nbasesInView) { super(); setBackground(Color.white); this.bam = bam; this.nbasesInView = nbasesInView; if(reference != null) { EntryGroup entryGroup = new SimpleEntryGroup(); try { getEntry(reference,entryGroup); } catch (NoSequenceException e) { e.printStackTrace(); } } try { readHeaderPicard(); } catch(java.lang.UnsupportedClassVersionError err) { JOptionPane.showMessageDialog(null, "This requires Java 1.6 or higher.", "Check Java Version", JOptionPane.WARNING_MESSAGE); } 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(""); } public String getToolTipText() { if(mouseOverSAMRecord == null) return null; String msg = mouseOverSAMRecord.getReadName() + "\n" + mouseOverSAMRecord.getAlignmentStart() + ".." + mouseOverSAMRecord.getAlignmentEnd() + "\nisize=" + mouseOverSAMRecord.getInferredInsertSize() + "\nmapq=" + mouseOverSAMRecord.getMappingQuality()+"\nrname="+ mouseOverSAMRecord.getReferenceName(); if( mouseOverSAMRecord.getReadPairedFlag() && mouseOverSAMRecord.getProperPairFlag() && !mouseOverSAMRecord.getMateUnmappedFlag()) { msg = msg + "\nstrand (read/mate): "+ (mouseOverSAMRecord.getReadNegativeStrandFlag() ? "-" : "+")+" / "+ (mouseOverSAMRecord.getMateNegativeStrandFlag() ? "-" : "+"); } else msg = msg + "\nstrand (read/mate): "+ (mouseOverSAMRecord.getReadNegativeStrandFlag() ? "-" : "+"); if(msg != null && mouseOverInsertion != null) msg = msg + "\nInsertion at:" +mouseOverInsertion; return msg; } /* private void readHeader() { String samtoolCmd = ""; if(System.getProperty("samtoolDir") != null) samtoolCmd = System.getProperty("samtoolDir"); String cmd[] = { samtoolCmd+File.separator+"samtools", "view", "-H", bam }; RunSamTools samtools = new RunSamTools(cmd, null, null, null); if(samtools.getProcessStderr() != null) System.out.println(samtools.getProcessStderr()); String header = samtools.getProcessStdout(); StringReader samReader = new StringReader(header); BufferedReader buff = new BufferedReader(samReader); String line; try { while((line = buff.readLine()) != null) { if(line.indexOf("LN:") > -1) { String parts[] = line.split("\t"); String name = ""; int seqLength = 0; for(int i=0; i<parts.length; i++) { if(parts[i].startsWith("LN:")) seqLength = Integer.parseInt( parts[i].substring(3) ); else if(parts[i].startsWith("SN:")) name = parts[i].substring(3); } seqLengths.put(name, seqLength); seqNames.add(name); } } } catch (IOException e) { e.printStackTrace(); } } private void readFromBam(int start, int end) { String refName = (String) combo.getSelectedItem(); String samtoolCmd = ""; if(System.getProperty("samtoolDir") != null) samtoolCmd = System.getProperty("samtoolDir"); String cmd[] = { samtoolCmd+File.separator+"samtools", "view", bam, refName+":"+start+"-"+end }; for(int i=0; i<cmd.length;i++) System.out.print(cmd[i]+" "); System.out.println(); if(readsInView == null) readsInView = new Vector<SAMRecord>(); else readsInView.clear(); RunSamTools samtools = new RunSamTools(cmd, null, null, readsInView); if(samtools.getProcessStderr() != null) System.out.println(samtools.getProcessStderr()); samtools.waitForStdout(); }*/ private void readHeaderPicard() { File bamFile = new File(bam); File indexFile = new File(bam+".bai"); final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile); SAMFileHeader header = inputSam.getFileHeader(); List<SAMSequenceRecord> readGroups = header.getSequenceDictionary().getSequences(); for(int i=0; i<readGroups.size(); i++) { seqLengths.put(readGroups.get(i).getSequenceName(), readGroups.get(i).getSequenceLength()); seqNames.add(readGroups.get(i).getSequenceName()); } inputSam.close(); } /** * Read a SAM or BAM file. */ private void readFromBamPicard(int start, int end) { // Open the input file. Automatically detects whether input is SAM or BAM // and delegates to a reader implementation for the appropriate format. File bamFile = new File(bam); File indexFile = new File(bam+".bai"); final SAMFileReader inputSam = new SAMFileReader(bamFile, indexFile); inputSam.setValidationStringency(ValidationStringency.SILENT); if(readsInView == null) readsInView = new Vector<SAMRecord>(); else readsInView.clear(); if(concatSequences) { int len = 0; int lastLen = 1; for(int i=0; i<seqNames.size(); i++) { int thisLength = seqLengths.get(seqNames.get(i)); len += thisLength; if( (lastLen >= start && lastLen < end) || (len >= start && len < end) || (start >= lastLen && start < len) || (end >= lastLen && end < len) ) { int offset = getSequenceOffset(seqNames.get(i)); int thisStart = start - offset; if(thisStart < 1) thisStart = 1; int thisEnd = end - offset; if(thisEnd > thisLength) thisEnd = thisLength; //System.out.println("READ "+seqNames.get(i)+" "+thisStart+".."+thisEnd); iterateOverBam(inputSam, seqNames.get(i), thisStart, thisEnd); } lastLen = len; } } else { String refName = (String) combo.getSelectedItem(); iterateOverBam(inputSam, refName, start, end); } inputSam.close(); //System.out.println("readFromBamPicard "+start+".."+end); } /** * 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, String refName, int start, int end) { CloseableIterator<SAMRecord> it = inputSam.queryOverlapping(refName, start, end); MemoryMXBean memory = ManagementFactory.getMemoryMXBean(); int checkMemAfter = 8000; int cnt = 0; while ( it.hasNext() ) { try { cnt++; SAMRecord samRecord = it.next(); if( samRecordFlagPredicate == null || !samRecordFlagPredicate.testPredicate(samRecord)) { if(samRecordMapQPredicate == null || samRecordMapQPredicate.testPredicate(samRecord)) readsInView.add(samRecord); } 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(heapFraction > 0.97) { popFrame.show( "Over 97 % of the maximum memory\nlimit ("+ (memory.getHeapMemoryUsage().getMax()/1000000.f)+" Mb).\n"+ "Zoom in or consider increasing the\nmemory for this application.", mainPanel, 15000); break; } } } catch(Exception e) { System.out.println(e.getMessage()); } } it.close(); } private int getSequenceLength() { if(concatSequences) { int len = 0; for(int i=0; i<seqNames.size(); i++) len += seqLengths.get(seqNames.get(i)); 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(!concatSequences) 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)); } } return offsetLengths.get(refName); } /** * Override */ protected void paintComponent(Graphics g) { super.paintComponent(g); Graphics2D g2 = (Graphics2D)g; mouseOverSAMRecord = null; int seqLength = getSequenceLength(); float pixPerBase = getPixPerBaseByWidth(); 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; } boolean changeToStackView = false; MemoryMXBean memory = ManagementFactory.getMemoryMXBean(); if(laststart != start || lastend != end) { if(!waitingFrame.isVisible()) waitingFrame.showWaiting("loading...", mainPanel); synchronized (this) { try { float heapFractionUsedBefore = (float) ((float) memory.getHeapMemoryUsage().getUsed() / (float) memory.getHeapMemoryUsage().getMax()); readFromBamPicard(start, end); 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) { checkBoxStackView.setSelected(true); isStackView = true; changeToStackView = true; } if ((!isStackView && !isStrandStackView) || pixPerBase * 1.08f >= ALIGNMENT_PIX_PER_BASE) { Collections.sort(readsInView, new SAMRecordComparator()); } } catch (OutOfMemoryError ome) { JOptionPane.showMessageDialog(this, "Out of Memory"); readsInView.clear(); return; } } } //System.out.println(start+".."+end+" " + // "sequence length = "+getSequenceLength()+ // " pixPerBase="+pixPerBase); laststart = start; lastend = end; if(showBaseAlignment) drawBaseAlignment(g2, seqLength, pixPerBase, start, end); 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.setStartAndEnd(start, end); coveragePanel.setPixPerBase(pixPerBase); coveragePanel.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; ruler.repaint(); int ypos = 0; String refSeq = null; int refSeqStart = start; end = start + ( mainPanel.getWidth() * ALIGNMENT_PIX_PER_BASE ); if(bases != null) { // draw the reference sequence ypos+=11; 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(); g2.setColor(lightGrey); g2.fillRect(0, ypos-11, mainPanel.getWidth(), 11); drawSelectionRange(g2, ALIGNMENT_PIX_PER_BASE, start, end); g2.setColor(Color.black); g2.drawString(refSeq, 0, ypos); } catch (OutOfRangeException e) { e.printStackTrace(); } } else drawSelectionRange(g2, ALIGNMENT_PIX_PER_BASE, start, end); 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; SAMRecord thisRead = readsInView.get(i); if (ypos < r.getMaxY() || ypos > r.getMinY()) drawSequence(g2, thisRead, ypos, refSeq, refSeqStart); drawn[i] = true; int thisEnd = thisRead.getAlignmentEnd(); if (thisEnd == 0) thisEnd = thisRead.getAlignmentStart() + thisRead.getReadLength(); for (int j = i + 1; j < nreads; j++) { if (!drawn[j]) { SAMRecord nextRead = readsInView.get(j); int nextStart = nextRead.getAlignmentStart(); if (nextStart > thisEnd + 1) { if (ypos < r.getMaxY() || ypos > r.getMinY()) drawSequence(g2, nextRead, ypos, refSeq, refSeqStart); drawn[j] = true; thisEnd = nextRead.getAlignmentEnd(); if (thisEnd == 0) thisEnd = nextStart + nextRead.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, SAMRecord samRecord, int ypos, String refSeq, int refSeqStart) { 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); 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); } // 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(deepPink); int xscreen = refPos*ALIGNMENT_PIX_PER_BASE; insertions.put(xscreen, (samRecord.getAlignmentStart()+len-2)+" "+ 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.getReadName().equals(samRecord.getReadName())) { refPos = blocks.get(0).getReferenceStart()+offset-refSeqStart; int xstart = refPos*ALIGNMENT_PIX_PER_BASE; int width = len*ALIGNMENT_PIX_PER_BASE; g2.setColor(Color.red); g2.drawRect(xstart, ypos-BASE_HEIGHT, width, BASE_HEIGHT); } if(lastMousePoint != null) { refPos = blocks.get(0).getReferenceStart()+offset-refSeqStart; int xstart = refPos*ALIGNMENT_PIX_PER_BASE; int xend = (refPos+len)*ALIGNMENT_PIX_PER_BASE; if(lastMousePoint.getY() > ypos-11 && lastMousePoint.getY() < ypos) if(lastMousePoint.getX() > xstart && lastMousePoint.getX() < xend) { mouseOverSAMRecord = samRecord; 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(darkGreen); else if (baseQuality < 30) g2.setColor(darkOrange); else g2.setColor(Color.black); } /** * Draw zoomed-out 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); if(showScale) drawScale(g2, start, end, pixPerBase, getHeight()); Stroke stroke = new BasicStroke (1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_ROUND); g2.setStroke(stroke); 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++) { SAMRecord samRecord = readsInView.get(i); SAMRecord samNextRecord = null; 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, samRecord, pixPerBase, ypos, baseAtStartOfView); } continue; } int ypos = getYPos(scaleHeight, Math.abs(samRecord.getInferredInsertSize())); if( (ypos > r.getMaxY() || ypos < r.getMinY()) && ypos > 0 ) continue; if(i < readsInView.size()-1) { samNextRecord = readsInView.get(++i); 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); drawTranslucentLine(g2, (int)((samRecord.getAlignmentEnd()-getBaseAtStartOfView())*pixPerBase), (int)((samNextRecord.getAlignmentStart()-getBaseAtStartOfView())*pixPerBase), ypos); } if( samRecord.getReadNegativeStrandFlag() && // strand of the query (1 for reverse) samNextRecord.getReadNegativeStrandFlag() ) g2.setColor(Color.red); else g2.setColor(Color.blue); drawRead(g2, samRecord, pixPerBase, ypos, baseAtStartOfView); drawRead(g2, samNextRecord, pixPerBase, ypos, baseAtStartOfView); } else { drawLoneRead(g2, samRecord, ypos, pixPerBase, baseAtStartOfView); i--; } } else { drawLoneRead(g2, samRecord, ypos, pixPerBase, baseAtStartOfView); } } drawYScale(g2, scaleHeight); } private int getYPos(int scaleHeight, int size) { int ypos; if(!logScale) ypos = (getHeight() - scaleHeight) - size; else { int logInfSize = (int)( Math.log(size) * 100); ypos = (getHeight() - scaleHeight) - logInfSize; } return ypos; } /** * 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, int seqLength, float pixPerBase, int start, int end) { drawSelectionRange(g2, pixPerBase,start, end); if(isShowScale()) drawScale(g2, start, end, pixPerBase, getHeight()); BasicStroke stroke = new BasicStroke( 1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_MITER); g2.setStroke(stroke); int scaleHeight; if(isShowScale()) scaleHeight = 15; else scaleHeight = 0; int ypos = (getHeight() - scaleHeight); int maxEnd = 0; int lstStart = 0; int lstEnd = 0; int baseAtStartOfView = getBaseAtStartOfView(); g2.setColor(Color.blue); Rectangle r = jspView.getViewport().getViewRect(); for(int i=0; i<readsInView.size(); i++) { SAMRecord samRecord = readsInView.get(i); int offset = getSequenceOffset(samRecord.getReferenceName()); int recordStart = samRecord.getAlignmentStart()+offset; int recordEnd = samRecord.getAlignmentEnd()+offset; if(lstStart != recordStart || lstEnd != recordEnd) { 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 = (getHeight() - scaleHeight)-2; maxEnd = recordEnd+2; } else ypos = ypos-2; } else g2.setColor(darkGreen); lstStart = recordStart; lstEnd = recordEnd; if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; drawRead(g2, samRecord, pixPerBase, ypos, baseAtStartOfView); } } /** * 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); BasicStroke stroke = new BasicStroke( 1.3f, BasicStroke.CAP_BUTT, BasicStroke.JOIN_MITER); int scaleHeight = 15; drawScale(g2, start, end, pixPerBase, ((getHeight()+scaleHeight)/2)); int ymid = (getHeight()/ 2); // positive strand drawStrand(g2, false, scaleHeight, ymid-(scaleHeight/2), -2, pixPerBase, stroke); // negative strand drawStrand(g2, true, scaleHeight, ymid+(scaleHeight/2), 2, pixPerBase, stroke); } private void drawStrand(Graphics2D g2, boolean isStrandNegative, int scaleHeight, int ymid, int ystep, float pixPerBase, Stroke stroke) { int ypos = (getHeight() - scaleHeight); int maxEnd = 0; int lstStart = 0; int lstEnd = 0; int baseAtStartOfView = getBaseAtStartOfView(); g2.setColor(Color.blue); Rectangle r = jspView.getViewport().getViewRect(); for(int i=0; i<readsInView.size(); i++) { SAMRecord samRecord = readsInView.get(i); if( samRecord.getReadNegativeStrandFlag() == isStrandNegative ) { int offset = getSequenceOffset(samRecord.getReferenceName()); int recordStart = samRecord.getAlignmentStart()+offset; int recordEnd = samRecord.getAlignmentEnd()+offset; if(lstStart != recordStart || lstEnd != recordEnd) { 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 = ymid + ystep; maxEnd = recordEnd+2; } else ypos = ypos + ystep; } else g2.setColor(darkGreen); lstStart = recordStart; lstEnd = recordEnd; if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; drawRead(g2, samRecord, pixPerBase, ypos, baseAtStartOfView); } } } /** * 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 drawPairedStackView(Graphics2D g2, int seqLength, float pixPerBase, int start, int end) { drawSelectionRange(g2, pixPerBase,start, end); if(isShowScale()) drawScale(g2, start, end, pixPerBase, getHeight()); Vector<PairedRead> pairedReads = new Vector<PairedRead>(); for(int i=0; i<readsInView.size(); i++) { SAMRecord samRecord = readsInView.get(i); if( !samRecord.getReadPairedFlag() || // read is not paired in sequencing samRecord.getMateUnmappedFlag() ) // mate is unmapped continue; SAMRecord samNextRecord = null; if(i < readsInView.size()-1) { samNextRecord = readsInView.get(++i); PairedRead pr = new PairedRead(); if(samRecord.getReadName().equals(samNextRecord.getReadName())) { if(samRecord.getAlignmentStart() < samNextRecord.getAlignmentStart()) { pr.sam1 = samRecord; pr.sam2 = samNextRecord; } else { pr.sam2 = samRecord; pr.sam1 = samNextRecord; } } else { --i; pr.sam1 = samRecord; 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); int scaleHeight; if(isShowScale()) scaleHeight = 15; else scaleHeight = 0; int ypos = getHeight() - scaleHeight - 3; int lastEnd = 0; int baseAtStartOfView = getBaseAtStartOfView(); Rectangle r = jspView.getViewport().getViewRect(); for(int i=0; i<pairedReads.size(); i++) { PairedRead pr = pairedReads.get(i); if(pr.sam1.getAlignmentStart() > lastEnd) { ypos = getHeight() - scaleHeight - 3; if(pr.sam2 != null) { lastEnd = pr.sam2.getAlignmentEnd(); } else lastEnd = pr.sam1.getAlignmentEnd(); } else ypos = ypos - 3; if(ypos > r.getMaxY() || ypos < r.getMinY()) continue; g2.setStroke(originalStroke); g2.setColor(Color.LIGHT_GRAY); if(pr.sam2 != null) { drawTranslucentJointedLine(g2, (int)((pr.sam1.getAlignmentEnd()-getBaseAtStartOfView())*pixPerBase), (int)((pr.sam2.getAlignmentStart()-getBaseAtStartOfView())*pixPerBase), ypos); } else { if(!pr.sam1.getMateUnmappedFlag() && pr.sam1.getMateReferenceName().equals(pr.sam1.getReferenceName())) { int prStart; if(pr.sam1.getAlignmentStart() > pr.sam1.getMateAlignmentStart()) prStart = pr.sam1.getAlignmentEnd(); else prStart = pr.sam1.getAlignmentStart(); drawTranslucentJointedLine(g2, (int)( (prStart-getBaseAtStartOfView())*pixPerBase), (int)( (pr.sam1.getMateAlignmentStart()-getBaseAtStartOfView())*pixPerBase), ypos); } } if( pr.sam1.getReadNegativeStrandFlag() && // strand of the query (1 for reverse) ( pr.sam2 != null && pr.sam2.getReadNegativeStrandFlag() ) ) g2.setColor(Color.red); else g2.setColor(Color.blue); drawRead(g2, pr.sam1, pixPerBase, ypos, baseAtStartOfView); if(pr.sam2 != null) drawRead(g2, pr.sam2, pixPerBase, ypos, baseAtStartOfView); } } /** * 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, SAMRecord samRecord, int ypos, float pixPerBase, int baseAtStartOfView) { 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(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(offTheTop) g2.setColor(darkOrange); else if(samRecord.getReadNegativeStrandFlag()) // strand of the query (1 for reverse) g2.setColor(Color.red); else g2.setColor(Color.blue); drawRead(g2, samRecord, pixPerBase, ypos, baseAtStartOfView); 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 */ private void drawRead(Graphics2D g2, SAMRecord thisRead, float pixPerBase, int ypos, int baseAtStartOfView) { int offset = getSequenceOffset(thisRead.getReferenceName()); int thisStart = thisRead.getAlignmentStart()+offset-baseAtStartOfView; int thisEnd = thisRead.getAlignmentEnd()+offset-baseAtStartOfView; if(highlightSAMRecord != null && highlightSAMRecord.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); } g2.drawLine((int)( thisStart * pixPerBase), ypos, (int)( 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 = thisRead; } } if (isSNPs) showSNPsOnReads(g2, thisRead, pixPerBase, ypos, offset); } /** * Highlight a selected range * @param g2 * @param pixPerBase * @param start * @param end */ private void drawSelectionRange(Graphics2D g2, float pixPerBase, int start, int end) { 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(Color.pink); 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 g2 * @param thisRead * @param pixPerBase * @param ypos */ private void showSNPsOnReads(Graphics2D g2, SAMRecord thisRead, float pixPerBase, int ypos, int offset) { int thisStart = thisRead.getAlignmentStart(); int thisEnd = thisRead.getAlignmentEnd(); // use alignment blocks of the contiguous alignment of // subsets of read bases to a reference sequence List<AlignmentBlock> blocks = thisRead.getAlignmentBlocks(); try { char[] refSeq = bases.getSubSequenceC( new Range(thisStart+offset, thisEnd+offset), Bases.FORWARD); byte[] readSeq = thisRead.getReadBases(); Color col = g2.getColor(); g2.setColor(Color.red); offset = offset - getBaseAtStartOfView(); for(int i=0; i<blocks.size(); i++) { AlignmentBlock block = blocks.get(i); for(int j=0; j<block.getLength(); j++) { int readPos = block.getReadStart()-1+j; int refPos = block.getReferenceStart()+j; if (Character.toUpperCase(refSeq[refPos-thisStart]) != readSeq[readPos]) { g2.drawLine((int) ((refPos+offset) * pixPerBase), ypos + 2, (int) ((refPos+offset) * pixPerBase), ypos - 2); } } } g2.setColor(col); } catch (OutOfRangeException e) { e.printStackTrace(); } } /** * Add the alignment view to the supplied <code>JPanel</code> in * a <code>JScrollPane</code>. * @param mainPanel panel to add the alignment to * @param autohide automatically hide the top panel containing the buttons */ public void addJamToPanel(final JPanel mainPanel, final JFrame frame, final boolean autohide, final FeatureDisplay feature_display) { this.mainPanel = mainPanel; final JComponent topPanel; if(feature_display != null) { this.feature_display = feature_display; this.selection = feature_display.getSelection(); topPanel = new JPanel(new FlowLayout(FlowLayout.LEADING, 0, 0)); } else { topPanel = new JMenuBar(); frame.setJMenuBar((JMenuBar)topPanel); JMenu fileMenu = new JMenu("File"); topPanel.add(fileMenu); JMenuItem readBam = new JMenuItem("Read BAM..."); fileMenu.add(readBam); readBam.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { String[] s = new String[0]; BamView.main(s); } }); 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); } }); } 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", autohide); 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 JComboBox(seqNames); combo.setEditable(false); combo.addItemListener(new ItemListener() { public void itemStateChanged(ItemEvent e) { laststart = -1; lastend = -1; 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("-"); 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.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); mainPanel.setPreferredSize(new Dimension(900, 400)); jspView = new JScrollPane(this, JScrollPane.VERTICAL_SCROLLBAR_AS_NEEDED, JScrollPane.HORIZONTAL_SCROLLBAR_NEVER); 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); 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(); } }); bottomPanel.add(scrollBar, BorderLayout.SOUTH); } mainPanel.add(bottomPanel, BorderLayout.SOUTH); coveragePanel.setPreferredSize(new Dimension(900, 100)); coveragePanel.setVisible(false); jspView.getVerticalScrollBar().setValue( jspView.getVerticalScrollBar().getMaximum()); jspView.getVerticalScrollBar().setUnitIncrement(maxUnitIncrement); if(feature_display == null) { 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; } } }); } addMouseListener(new PopupListener()); setFocusable(true); requestFocusInWindow(); } private void createMenus(JComponent menu) { JMenu viewMenu = new JMenu("Views"); ButtonGroup group = new ButtonGroup(); final JCheckBoxMenuItem checkBoxPairedStackView = new JCheckBoxMenuItem("Paired Stack View"); final JCheckBoxMenuItem checkBoxStrandStackView = new JCheckBoxMenuItem("Strand Stack View"); final JCheckBoxMenuItem checkIsizeStackView = new JCheckBoxMenuItem("Inferred Size View", true); checkBoxStackView.setFont(checkIsizeStackView.getFont()); baseQualityColour.setFont(checkIsizeStackView.getFont()); markInsertions.setFont(checkIsizeStackView.getFont()); group.add(checkBoxStackView); group.add(checkBoxPairedStackView); group.add(checkBoxStrandStackView); group.add(checkIsizeStackView); checkIsizeStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; lastend = -1; if(checkIsizeStackView.isSelected()) { isStackView = false; isPairedStackView = false; isStrandStackView = false; logMenuItem.setEnabled(true); } repaint(); } }); viewMenu.add(checkIsizeStackView); checkBoxStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; lastend = -1; isStackView = !isStackView; if(isStackView) { isPairedStackView = !isStackView; isStrandStackView = !isStackView; checkBoxPairedStackView.setSelected(!isStackView); checkBoxStrandStackView.setSelected(!isStackView); logMenuItem.setEnabled(false); } repaint(); } }); viewMenu.add(checkBoxStackView); checkBoxPairedStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; lastend = -1; isPairedStackView = !isPairedStackView; if(isPairedStackView) { isStackView = !isPairedStackView; isStrandStackView = !isPairedStackView; checkBoxStackView.setSelected(!isPairedStackView); checkBoxStrandStackView.setSelected(!isPairedStackView); logMenuItem.setEnabled(false); } repaint(); } }); viewMenu.add(checkBoxPairedStackView); checkBoxStrandStackView.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { laststart = -1; lastend = -1; isStrandStackView = !isStrandStackView; if(isStrandStackView) { isStackView = !isStrandStackView; isPairedStackView = !isStrandStackView; checkBoxStackView.setSelected(!isStrandStackView); checkBoxPairedStackView.setSelected(!isStrandStackView); setViewportMidPoint(); logMenuItem.setEnabled(false); } repaint(); } }); viewMenu.add(checkBoxStrandStackView); menu.add(viewMenu); JMenu showMenu = new JMenu("Show"); JCheckBoxMenuItem checkBoxSingle = new JCheckBoxMenuItem("Single Reads"); checkBoxSingle.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { repaint(); isSingle = !isSingle; } }); showMenu.add(checkBoxSingle); final JCheckBoxMenuItem checkBoxSNPs = new JCheckBoxMenuItem("SNPs"); 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); baseQualityColour.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { if(baseQualityColour.isSelected()) { checkBoxSNPs.setSelected(false); isSNPs = false; } repaint(); } }); showMenu.add(baseQualityColour); markInsertions.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { repaint(); } }); showMenu.add(markInsertions); showMenu.add(new JSeparator()); JCheckBoxMenuItem checkBoxCoverage = new JCheckBoxMenuItem("Coverage"); checkBoxCoverage.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { isCoverage = !isCoverage; coveragePanel.setVisible(isCoverage); repaint(); } }); showMenu.add(checkBoxCoverage); menu.add(showMenu); 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("Plot 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(checkIsizeStackView.getFont()); menu.add(logMenuItem); logMenuItem.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { logScale = logMenuItem.isSelected(); repaint(); } }); JMenuItem filter = new JMenuItem("Filter by Flag..."); menu.add(filter); filter.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { new SAMRecordFilter(BamView.this); } }); menu.add(new JSeparator()); } 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 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(pixPerBase*1.08f >= ALIGNMENT_PIX_PER_BASE) { 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) { jspView.setColumnHeaderView(null); if(!isStrandStackView) jspView.getVerticalScrollBar().setValue( jspView.getVerticalScrollBar().getMaximum()); 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 showScale; } public void setShowScale(boolean showScale) { this.showScale = showScale; } public JScrollPane getJspView() { return jspView; } public void setBases(Bases bases) { this.bases = bases; } /** * Remove JScrollPane border */ public void removeBorder() { Border empty = new EmptyBorder(0,0,0,0); jspView.setBorder(empty); } /** * 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 */ private 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(); } } private Selection getSelection() { return selection; } protected List<SAMRecord> getReadsInView() { return readsInView; } protected int getBasesInView() { return nbasesInView; } /** * Artemis event notification */ public void displayAdjustmentValueChanged(final DisplayAdjustmentEvent event) { 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; lastend = -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; int start; int end; public Ruler() { super(); setPreferredSize(new Dimension(mainPanel.getWidth(), 15)); setBackground(Color.white); setFont(getFont().deriveFont(11.f)); } 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+=10) { int xpos = (i-start)*ALIGNMENT_PIX_PER_BASE; g2.drawString(Integer.toString(i), xpos, ypos); xpos+=(ALIGNMENT_PIX_PER_BASE/2); g2.drawLine(xpos, ypos+1, xpos, ypos+5); } } } /** * Popup menu listener */ class PopupListener extends MouseAdapter { JMenuItem gotoMateMenuItem; JMenuItem showDetails; 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) 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()) { if(popup == null) { popup = new JPopupMenu(); createMenus(popup); } if(gotoMateMenuItem != null) popup.remove(gotoMateMenuItem); if(showDetails != null) popup.remove(showDetails); if( mouseOverSAMRecord != null && mouseOverSAMRecord.getReadPairedFlag() && !mouseOverSAMRecord.getMateUnmappedFlag() ) { final SAMRecord thisSAMRecord = mouseOverSAMRecord; gotoMateMenuItem = new JMenuItem("Go to mate of : "+ thisSAMRecord.getReadName()); gotoMateMenuItem.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { String name = thisSAMRecord.getMateReferenceName(); if(name.equals("=")) name = thisSAMRecord.getReferenceName(); int offset = getSequenceOffset(name); if(feature_display != null) feature_display.makeBaseVisible( thisSAMRecord.getMateAlignmentStart()+offset); else scrollBar.setValue( thisSAMRecord.getMateAlignmentStart()+offset- (nbasesInView/2)); highlightSAMRecord = thisSAMRecord; } }); popup.add(gotoMateMenuItem); } if( mouseOverSAMRecord != null) { final SAMRecord thisSAMRecord = mouseOverSAMRecord; showDetails = new JMenuItem("Show details of : "+ thisSAMRecord.getReadName()); showDetails.addActionListener(new ActionListener() { public void actionPerformed(ActionEvent e) { FileViewer viewDetail = new FileViewer(thisSAMRecord.getReadName(), true, false); appendToDetailView(thisSAMRecord, viewDetail); } }); popup.add(showDetails); } popup.show(e.getComponent(), e.getX(), e.getY()); } } } private void appendToDetailView(SAMRecord thisSAMRecord, FileViewer viewDetail) { 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); if(thisSAMRecord.getProperPairFlag() && !thisSAMRecord.getMateUnmappedFlag()) { viewDetail.appendString("Mate Reference Name "+thisSAMRecord.getMateReferenceName()+"\n", Level.DEBUG); viewDetail.appendString("Mate Start Coordinate "+thisSAMRecord.getMateAlignmentStart()+"\n", Level.DEBUG); viewDetail.appendString("Strand (read/mate) "+ (thisSAMRecord.getReadNegativeStrandFlag() ? "-" : "+")+" / "+ (thisSAMRecord.getMateNegativeStrandFlag() ? "-" : "+"), Level.DEBUG); } else { viewDetail.appendString("Strand (read) "+ (thisSAMRecord.getReadNegativeStrandFlag() ? "-" : "+"), Level.DEBUG); } viewDetail.appendString("\n\nCigar String "+thisSAMRecord.getCigarString(), Level.DEBUG); viewDetail.appendString("\n\nFlags:", Level.INFO); viewDetail.appendString("\nDuplicate Read "+ (thisSAMRecord.getDuplicateReadFlag() ? "yes" : "no"), Level.DEBUG); 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); viewDetail.appendString("\nSecond Of Pair "+ (thisSAMRecord.getSecondOfPairFlag() ? "yes" : "no"), Level.DEBUG); viewDetail.appendString("\n\nRead Bases:\n", Level.INFO); viewDetail.appendString(new String(thisSAMRecord.getReadBases()), Level.DEBUG); } protected SAMRecordFlagPredicate getSamRecordFlagPredicate() { return samRecordFlagPredicate; } protected void setSamRecordFlagPredicate( SAMRecordFlagPredicate 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; } class PairedRead { SAMRecord sam1; SAMRecord sam2; } public static void main(String[] args) { String bam; String reference = null; if(args.length == 0) { FileSelectionPanel fileSelection = new FileSelectionPanel(); fileSelection.showPanel(); bam = fileSelection.getBamFile(); reference = fileSelection.getReferenceFile(); if(reference == null || reference.equals("")) reference = null; } else bam = args[0]; int nbasesInView = 1000; for(int i=0;i<args.length; i++) { if(args[i].equals("-a")) bam = args[++i]; else if(args[i].equals("-r")) reference = args[++i]; else if(args[i].equals("-v")) nbasesInView = Integer.parseInt(args[++i]); else if(args[i].equals("-s")) System.setProperty("samtoolDir", args[++i]); 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("-v\t number of bases to display in the view (optional)"); /*System.out.println("-s\t samtool directory");*/ System.exit(0); } } final BamView view = new BamView(bam, reference, nbasesInView); JFrame frame = new JFrame("BamView"); // 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){} }); view.addJamToPanel((JPanel)frame.getContentPane(), frame, false, null); frame.pack(); view.jspView.getVerticalScrollBar().setValue( view.jspView.getVerticalScrollBar().getMaximum()); frame.setVisible(true); } }