public void set(MouseEvent e, SAMRecord sr) {
      if (sr == null) return;
      StringBuffer text = new StringBuffer();
      text.append("<html>");

      if (sr != null) {
        text.append(
            MessageManager.getString("shortreadtrack.name") + " " + sr.getReadName() + "<br/>");
        text.append(
            MessageManager.getString("shortreadtrack.len") + " " + sr.getReadLength() + "<br/>");
        text.append(
            MessageManager.getString("shortreadtrack.cigar") + " " + sr.getCigarString() + "<br/>");
        text.append(
            MessageManager.getString("shortreadtrack.sequence")
                + " "
                + rerun(sr.getReadString())
                + "<br/>");
        text.append(
            MessageManager.getString("shortreadtrack.paired")
                + " "
                + sr.getReadPairedFlag()
                + "<br/>");
        if (sr.getReadPairedFlag()) {
          if (!sr.getMateUnmappedFlag())
            text.append(
                MessageManager.getString("shortreadtrack.mate")
                    + " "
                    + sr.getMateReferenceName()
                    + ":"
                    + sr.getMateAlignmentStart()
                    + "<br/>");
          else text.append(MessageManager.getString("shortreadtrack.mate_missing") + "<br/>");
          text.append(
              MessageManager.getString("shortreadtrack.second") + " " + sr.getFirstOfPairFlag());
        }
        // text.append("<br/>");
      }
      text.append("</html>");
      if (!text.toString().equals(floater.getText())) {
        floater.setText(text.toString());
        this.pack();
      }
      setLocation(e.getXOnScreen() + 5, e.getYOnScreen() + 5);

      if (!isVisible()) {
        setVisible(true);
      }
    }
Beispiel #2
0
  public static void align(
      Graph graph,
      SAMRecord rec,
      Node recNode,
      ReferenceSequence sequence,
      SAMProgramRecord programRecord,
      int offset,
      AlleleCoverageCutoffs alleleCoverageCutoffs,
      boolean correctBases,
      boolean useSequenceQualities,
      int MAXIMUM_TOTAL_COVERAGE,
      int MAX_HEAP_SIZE)
      throws Exception {

    int i;
    AlignHeapNode curAlignHeapNode = null;
    AlignHeapNode nextAlignHeapNode = null;
    AlignHeapNode bestAlignHeapNode = null;
    AlignHeap heap = null;
    String read = null; // could be cs
    String readBases = null; // always nt
    String qualities = null; // could be cq
    SRMAUtil.Space space = SRMAUtil.Space.NTSPACE;
    ListIterator<NodeRecord> iter = null;
    AlignHeapNodeComparator comp = null;
    int alignmentStart = -1;
    int numStartNodesAdded = 0;
    boolean strand = rec.getReadNegativeStrandFlag(); // false -> forward, true -> reverse
    String softClipStartBases = null;
    String softClipStartQualities = null;
    String softClipEndBases = null;
    String softClipEndQualities = null;

    // Debugging stuff
    String readName = rec.getReadName();

    assert SRMAUtil.Space.COLORSPACE != space;

    // Get space
    read = (String) rec.getAttribute("CS");
    if (null == read) {
      // Use base space
      space = SRMAUtil.Space.NTSPACE;
    } else {
      // assumes CS and CQ are always in sequencing order
      space = SRMAUtil.Space.COLORSPACE;
    }

    // Get read and qualities
    if (space == SRMAUtil.Space.NTSPACE) {
      byte tmpRead[] = rec.getReadString().getBytes();
      byte tmpQualities[] = rec.getBaseQualityString().getBytes();
      // Reverse once
      if (strand) { // reverse
        SAMRecordUtil.reverseArray(tmpRead);
        SAMRecordUtil.reverseArray(tmpQualities);
      }
      read = new String(tmpRead);
      readBases = new String(tmpRead);
      qualities = new String(tmpQualities);
      // Reverse again
      if (strand) { // reverse
        SAMRecordUtil.reverseArray(tmpRead);
        SAMRecordUtil.reverseArray(tmpQualities);
      }
    } else {
      byte tmpRead[] = rec.getReadString().getBytes();
      // Reverse once
      if (strand) { // reverse
        SAMRecordUtil.reverseArray(tmpRead);
      }
      readBases = new String(tmpRead);
      // Reverse again
      if (strand) { // reverse
        SAMRecordUtil.reverseArray(tmpRead);
      }
      read = SRMAUtil.normalizeColorSpaceRead(read);
      qualities = (String) rec.getAttribute("CQ");
      // Some aligners include a quality value for the adapter.  A quality value
      // IMHO should not be given for an unobserved (assumed) peice of data.  Trim
      // the first quality in this case
      if (qualities.length() == 1 + read.length()) { // trim the first quality
        qualities = qualities.substring(1);
      }
    }
    // Reverse back
    if (readBases.length() <= 0) {
      throw new Exception("Error.  The current alignment has no bases.");
    }
    if (read.length() <= 0) {
      throw new Exception("Error.  The current alignment has no bases.");
    }
    if (qualities.length() <= 0) {
      throw new Exception("Error.  The current alignment has no qualities.");
    }
    if (readBases.length() != read.length()) {
      if (space == SRMAUtil.Space.COLORSPACE) {
        throw new Exception(
            "Error.  The current alignment's read bases length does not match the length of the colors in the CS tag ["
                + rec.getReadName()
                + "].");
      } else {
        throw new Exception("Error.  Internal error: readBases.length() != read.length()");
      }
    }

    // Deal with soft-clipping
    // - save the soft clipped sequence for latter
    {
      List<CigarElement> cigarElements = null;

      cigarElements = rec.getCigar().getCigarElements();
      CigarElement e1 = cigarElements.get(0); // first
      CigarElement e2 = cigarElements.get(cigarElements.size() - 1); // last

      // Soft-clipped
      if (CigarOperator.S == e1.getOperator()) {
        if (space == SRMAUtil.Space.COLORSPACE) {
          throw new Exception(
              "Error.  Soft clipping with color-space data not currently supported.");
        }
        int l = e1.getLength();
        if (strand) { // reverse
          softClipStartBases = readBases.substring(readBases.length() - l);
          softClipStartQualities = qualities.substring(qualities.length() - l);
          readBases = readBases.substring(0, readBases.length() - l);
          read = read.substring(0, read.length() - l);
          qualities = qualities.substring(0, qualities.length() - l);
        } else {
          softClipStartBases = readBases.substring(0, l - 1);
          softClipStartQualities = qualities.substring(0, l - 1);
          readBases = readBases.substring(l);
          read = read.substring(l);
          qualities = qualities.substring(l);
        }
      }
      if (CigarOperator.S == e2.getOperator()) {
        if (space == SRMAUtil.Space.COLORSPACE) {
          throw new Exception(
              "Error.  Soft clipping with color-space data not currently supported.");
        }
        int l = e2.getLength();
        if (strand) { // reverse
          softClipEndBases = readBases.substring(0, l - 1);
          softClipEndQualities = qualities.substring(0, l - 1);
          readBases = readBases.substring(l);
          read = read.substring(l);
          qualities = qualities.substring(l);
        } else {
          softClipEndBases = readBases.substring(readBases.length() - l);
          softClipEndQualities = qualities.substring(qualities.length() - l);
          readBases = readBases.substring(0, readBases.length() - l);
          read = read.substring(0, read.length() - l);
          qualities = qualities.substring(0, qualities.length() - l);
        }
      }
    }

    // Remove mate pair information
    Align.removeMateInfo(rec);

    comp =
        new AlignHeapNodeComparator(
            (strand) ? AlignHeap.HeapType.MAXHEAP : AlignHeap.HeapType.MINHEAP);

    // Bound by original alignment if possible
    bestAlignHeapNode =
        Align.boundWithOriginalAlignment(
            rec,
            graph,
            recNode,
            comp,
            strand,
            read,
            qualities,
            readBases,
            space,
            sequence,
            alleleCoverageCutoffs,
            useSequenceQualities,
            MAXIMUM_TOTAL_COVERAGE,
            MAX_HEAP_SIZE);

    /*
    System.err.println("readName="+rec.getReadName());
    if(null != bestAlignHeapNode) {
    System.err.println("\nFOUND BEST:" + rec.toString());
    }
    else {
    System.err.println("\nNOT FOUND (BEST): " + rec.toString());
    }
    Align.updateSAM(rec, programRecord, bestAlignHeapNode, space, read, qualities, softClipStartBases, softClipStartQualities, softClipEndBases, softClipEndQualities, strand, correctBases);
    return;
    */

    heap = new AlignHeap((strand) ? AlignHeap.HeapType.MAXHEAP : AlignHeap.HeapType.MINHEAP);

    // Add start nodes
    if (strand) { // reverse
      alignmentStart = rec.getAlignmentEnd();
      for (i = alignmentStart + offset; alignmentStart - offset <= i; i--) {
        int position = graph.getPriorityQueueIndexAtPositionOrBefore(i);
        PriorityQueue<Node> startNodeQueue = graph.getPriorityQueue(position);
        if (0 != position && null != startNodeQueue) {
          Iterator<Node> startNodeQueueIter = startNodeQueue.iterator();
          while (startNodeQueueIter.hasNext()) {
            Node startNode = startNodeQueueIter.next();
            int f = passFilters(graph, startNode, alleleCoverageCutoffs, MAXIMUM_TOTAL_COVERAGE);
            if (0 == f) {
              heap.add(
                  new AlignHeapNode(
                      null,
                      startNode,
                      startNode.coverage,
                      read.charAt(0),
                      qualities.charAt(0),
                      useSequenceQualities,
                      space));
            } else if (f < 0) {
              return;
            }
            if (startNode.position < i) {
              i = startNode.position;
            }
            numStartNodesAdded++;
          }
        }
      }
    } else {
      alignmentStart = rec.getAlignmentStart();
      for (i = alignmentStart - offset; i <= alignmentStart + offset; i++) {
        int position = graph.getPriorityQueueIndexAtPositionOrGreater(i);
        PriorityQueue<Node> startNodeQueue = graph.getPriorityQueue(position);
        if (0 != position && null != startNodeQueue) {
          Iterator<Node> startNodeQueueIter = startNodeQueue.iterator();
          while (startNodeQueueIter.hasNext()) {
            Node startNode = startNodeQueueIter.next();
            int f = passFilters(graph, startNode, alleleCoverageCutoffs, MAXIMUM_TOTAL_COVERAGE);
            if (0 == f) {
              heap.add(
                  new AlignHeapNode(
                      null,
                      startNode,
                      startNode.coverage,
                      read.charAt(0),
                      qualities.charAt(0),
                      useSequenceQualities,
                      space));
            } else if (f < 0) {
              return;
            }
            if (i < startNode.position) {
              i = startNode.position;
            }
            numStartNodesAdded++;
          }
        }
      }
    }
    if (numStartNodesAdded == 0) {
      throw new Exception("Did not add any start nodes!");
    }

    // Get first node off the heap
    curAlignHeapNode = heap.poll();

    while (null != curAlignHeapNode) {

      if (MAX_HEAP_SIZE <= heap.size()) {
        // too many to consider
        return;
      }

      // System.err.println("strand:" + strand + "\tsize:" + heap.size() + "\talignmentStart:" +
      // alignmentStart + "\toffset:" + offset + "\treadOffset:" + curAlignHeapNode.readOffset);
      // System.err.print("size:" + heap.size() + ":" + curAlignHeapNode.readOffset + ":" +
      // curAlignHeapNode.score + ":" + curAlignHeapNode.alleleCoverageSum + ":" +
      // curAlignHeapNode.startPosition + "\t");
      // curAlignHeapNode.node.print(System.err);
      // System.err.print("\rposition:" + curAlignHeapNode.node.position + "\treadOffset:" +
      // curAlignHeapNode.readOffset);

      // Remove all non-insertions with the same contig/pos/read-offset/type/base and lower score
      nextAlignHeapNode = heap.peek();
      while (Node.INSERTION != curAlignHeapNode.node.type
          && null != nextAlignHeapNode
          && 0 == comp.compare(curAlignHeapNode, nextAlignHeapNode)) {
        if (curAlignHeapNode.score < nextAlignHeapNode.score
            || (curAlignHeapNode.score == nextAlignHeapNode.score
                && curAlignHeapNode.alleleCoverageSum < nextAlignHeapNode.alleleCoverageSum)) {
          // Update current node
          curAlignHeapNode = heap.poll();
        } else {
          // Ignore next node
          heap.poll();
        }
        nextAlignHeapNode = heap.peek();
      }
      nextAlignHeapNode = null;

      // Check if the alignment is complete
      if (curAlignHeapNode.readOffset == read.length() - 1) {
        // All read bases examined, store if has the best alignment.

        // System.err.print(curAlignHeapNode.alleleCoverageSum + ":" + curAlignHeapNode.score +
        // ":");
        // System.err.print(curAlignHeapNode.startPosition + ":");
        // curAlignHeapNode.node.print(System.err);

        if (null == bestAlignHeapNode
            || bestAlignHeapNode.score < curAlignHeapNode.score
            || (bestAlignHeapNode.score == curAlignHeapNode.score
                && bestAlignHeapNode.alleleCoverageSum < curAlignHeapNode.alleleCoverageSum)) {
          bestAlignHeapNode = curAlignHeapNode;
        }
      } else if (null != bestAlignHeapNode && curAlignHeapNode.score < bestAlignHeapNode.score) {
        // ignore, under the assumption that scores can only become more negative.
      } else {
        if (strand) { // reverse
          // Go to all the "prev" nodes
          iter = curAlignHeapNode.node.prev.listIterator();
        } else { // forward
          // Go to all "next" nodes
          iter = curAlignHeapNode.node.next.listIterator();
        }
        while (iter.hasNext()) {
          NodeRecord next = iter.next();
          int f =
              passFilters(
                  graph, next.node, next.coverage, alleleCoverageCutoffs, MAXIMUM_TOTAL_COVERAGE);
          if (0 == f) {
            heap.add(
                new AlignHeapNode(
                    curAlignHeapNode,
                    next.node,
                    next.coverage,
                    read.charAt(curAlignHeapNode.readOffset + 1),
                    qualities.charAt(curAlignHeapNode.readOffset + 1),
                    useSequenceQualities,
                    space));
          } else if (f < 0) {
            return;
          }
        }
        iter = null;
      }
      // Get next node
      curAlignHeapNode = heap.poll();
    }

    // Recover alignment
    Align.updateSAM(
        rec,
        sequence,
        programRecord,
        bestAlignHeapNode,
        space,
        read,
        qualities,
        softClipStartBases,
        softClipStartQualities,
        softClipEndBases,
        softClipEndQualities,
        strand,
        correctBases);
  }
Beispiel #3
0
  @Override
  public int doWork(String[] args) {
    boolean repair_missing_read = false;
    SortingCollectionFactory<MappedFastq> sortingFactory =
        new SortingCollectionFactory<MappedFastq>();
    File forwardFile = null;
    File reverseFile = null;
    com.github.lindenb.jvarkit.util.cli.GetOpt opt =
        new com.github.lindenb.jvarkit.util.cli.GetOpt();
    int c;

    sortingFactory.setComponentType(MappedFastq.class);
    sortingFactory.setCodec(new MappedFastqCodec());
    sortingFactory.setComparator(new MappedFastqComparator());

    while ((c = opt.getopt(args, super.getGetOptDefault() + "F:R:N:r")) != -1) {
      switch (c) {
        case 'F':
          forwardFile = new File(opt.getOptArg());
          break;
        case 'R':
          reverseFile = new File(opt.getOptArg());
          break;
        case 't':
          addTmpDirectory(new File(opt.getOptArg()));
          break;
        case 'N':
          sortingFactory.setMaxRecordsInRAM(Math.max(Integer.parseInt(opt.getOptArg()), 100));
          break;
        case 'r':
          repair_missing_read = true;
          break;
        case ':':
          System.err.println("Missing argument for option -" + opt.getOptOpt());
          return -1;
        default:
          {
            switch (handleOtherOptions(c, opt, args)) {
              case EXIT_FAILURE:
                return -1;
              case EXIT_SUCCESS:
                return 0;
              default:
                break;
            }
          }
      }
    }
    SAMFileReader sfr = null;
    SortingCollection<MappedFastq> fastqCollection = null;
    try {
      sortingFactory.setTmpDirs(this.getTmpDirectories());
      fastqCollection = sortingFactory.make();
      fastqCollection.setDestructiveIteration(true);
      boolean found_single = false;
      boolean found_paired = false;
      long non_primary_alignmaned_flag = 0L;

      if (opt.getOptInd() == args.length) {
        info("Reading from stdin");
        sfr = new SAMFileReader(System.in);
      } else if (opt.getOptInd() + 1 == args.length) {
        String filename = args[opt.getOptInd()];
        sfr = new SAMFileReader(new File(filename));
      } else {
        error(getMessageBundle("illegal.number.of.arguments"));
        return -1;
      }
      sfr.setValidationStringency(ValidationStringency.LENIENT);
      SAMRecordIterator iter = sfr.iterator();
      SAMSequenceDictionaryProgress progress =
          new SAMSequenceDictionaryProgress(sfr.getFileHeader().getSequenceDictionary());
      while (iter.hasNext()) {
        SAMRecord rec = iter.next();
        progress.watch(rec);

        if (rec.isSecondaryOrSupplementary()) {
          if (non_primary_alignmaned_flag == 0) {
            warning("SKIPPING NON-PRIMARY " + (non_primary_alignmaned_flag + 1) + " ALIGNMENTS");
          }
          non_primary_alignmaned_flag++;
          continue;
        }

        MappedFastq m = new MappedFastq();
        m.name = rec.getReadName();
        if (m.name == null) m.name = "";
        m.hash = m.name.hashCode();
        m.seq = rec.getReadString();

        if (m.seq.equals(SAMRecord.NULL_SEQUENCE_STRING)) m.seq = "";
        m.qual = rec.getBaseQualityString();
        if (m.qual.equals(SAMRecord.NULL_QUALS_STRING)) m.qual = "";
        if (!rec.getReadUnmappedFlag() && rec.getReadNegativeStrandFlag()) {
          m.seq = AcidNucleics.reverseComplement(m.seq);
          m.qual = new StringBuilder(m.qual).reverse().toString();
        }
        if (m.seq.length() != m.qual.length()) {
          error("length(seq)!=length(qual) in " + m.name);
          continue;
        }
        if (m.seq.isEmpty() && m.qual.isEmpty()) {
          m.seq = "N";
          m.qual = "#";
        }

        if (rec.getReadPairedFlag()) {
          found_paired = true;
          if (found_single) {
            sfr.close();
            throw new PicardException("input is a mix of paired/singled reads");
          }
          m.side = (byte) (rec.getSecondOfPairFlag() ? 2 : 1);
        } else {
          found_single = true;
          if (found_paired) {
            sfr.close();
            throw new PicardException("input is a mix of paired/singled reads");
          }
          m.side = (byte) 0;
        }
        fastqCollection.add(m);
      }
      iter.close();
      CloserUtil.close(iter);
      CloserUtil.close(sfr);
      progress.finish();

      fastqCollection.doneAdding();
      info("Done reading.");

      if (found_paired) {
        FastqWriter fqw1 = null;
        FastqWriter fqw2 = null;
        if (forwardFile != null) {
          info("Writing to " + forwardFile);
          fqw1 = new BasicFastqWriter(forwardFile);
        } else {
          info("Writing to stdout");
          fqw1 = new BasicFastqWriter(new PrintStream(System.out));
        }
        if (reverseFile != null) {
          info("Writing to " + reverseFile);
          fqw2 = new BasicFastqWriter(reverseFile);
        } else {
          info("Writing to interlaced stdout");
          fqw2 = fqw1;
        }
        List<MappedFastq> row = new ArrayList<MappedFastq>();
        CloseableIterator<MappedFastq> r = fastqCollection.iterator();
        for (; ; ) {
          MappedFastq curr = null;
          if (r.hasNext()) curr = r.next();
          if (curr == null || (!row.isEmpty() && !row.get(0).name.equals(curr.name))) {
            if (!row.isEmpty()) {
              if (row.size() > 2) {
                warning("WTF :" + row);
              }
              boolean found_F = false;
              boolean found_R = false;
              for (MappedFastq m : row) {
                switch ((int) m.side) {
                  case 1:
                    if (found_F)
                      throw new PicardException("two forward reads found for " + row.get(0).name);
                    found_F = true;
                    echo(fqw1, m);
                    break;
                  case 2:
                    if (found_R)
                      throw new PicardException("two reverse reads found for " + row.get(0).name);
                    found_R = true;
                    echo(fqw2, m);
                    break;
                  default:
                    throw new IllegalStateException("uh???");
                }
              }
              if (!found_F) {
                if (repair_missing_read) {
                  warning("forward not found for " + row.get(0));
                  MappedFastq pad = new MappedFastq();
                  pad.side = (byte) 1;
                  pad.name = row.get(0).name;
                  pad.seq = "N";
                  pad.qual = "#";
                  echo(fqw1, pad);
                } else {
                  throw new PicardException("forward not found for " + row);
                }
              }
              if (!found_R) {
                if (repair_missing_read) {
                  warning("reverse not found for " + row.get(0));
                  MappedFastq pad = new MappedFastq();
                  pad.side = (byte) 2;
                  pad.name = row.get(0).name;
                  pad.seq = "N";
                  pad.qual = "#";
                  echo(fqw2, pad);
                } else {
                  throw new PicardException("reverse not found for " + row);
                }
              }
            }
            if (curr == null) break;
            row.clear();
          }
          row.add(curr);
        }
        r.close();
        fqw1.close();
        fqw2.close();
      } else if (found_single) {
        FastqWriter fqw1 = null;
        if (forwardFile != null) {
          info("Writing to " + forwardFile);
          fqw1 = new BasicFastqWriter(forwardFile);
        } else {
          info("Writing to stdout");
          fqw1 = new BasicFastqWriter(new PrintStream(System.out));
        }

        CloseableIterator<MappedFastq> r = fastqCollection.iterator();
        while (r.hasNext()) {
          echo(fqw1, r.next());
        }
        r.close();
        fqw1.close();
      }
      return 0;
    } catch (Exception err) {
      error(err);
      return -1;
    } finally {
      if (fastqCollection != null) fastqCollection.cleanup();
    }
  }