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);
      }
    }
    public void acceptRecord(final SAMRecordAndReference args) {
      final SAMRecord rec = args.getSamRecord();
      final ReferenceSequence ref = args.getReferenceSequence();

      if (rec.getReadPairedFlag()) {
        if (rec.getFirstOfPairFlag()) {
          firstOfPairCollector.addRecord(rec, ref);
        } else {
          secondOfPairCollector.addRecord(rec, ref);
        }

        pairCollector.addRecord(rec, ref);
      } else {
        unpairedCollector.addRecord(rec, ref);
      }
    }
Beispiel #3
0
  private static void removeMateInfo(SAMRecord rec) {
    if (rec.getReadPairedFlag()) {
      // Remove all information of its mate

      // flag
      rec.setProperPairFlag(false); // not paired any more
      rec.setMateUnmappedFlag(false);
      rec.setMateNegativeStrandFlag(false);

      // entries
      rec.setMateReferenceIndex(SAMRecord.NO_ALIGNMENT_REFERENCE_INDEX);
      rec.setMateAlignmentStart(0);
      rec.setInferredInsertSize(0);

      // TODO: remove tags and values that are mate pair inclined.
    }
  }
      private void collectReadData(final SAMRecord record, final ReferenceSequence ref) {
        metrics.TOTAL_READS++;
        readLengthHistogram.increment(record.getReadBases().length);

        if (!record.getReadFailsVendorQualityCheckFlag()) {
          metrics.PF_READS++;
          if (isNoiseRead(record)) metrics.PF_NOISE_READS++;

          if (record.getReadUnmappedFlag()) {
            // If the read is unmapped see if it's adapter sequence
            final byte[] readBases = record.getReadBases();
            if (!(record instanceof BAMRecord)) StringUtil.toUpperCase(readBases);

            if (isAdapterSequence(readBases)) {
              this.adapterReads++;
            }
          } else if (doRefMetrics) {
            metrics.PF_READS_ALIGNED++;
            if (!record.getReadNegativeStrandFlag()) numPositiveStrand++;

            if (record.getReadPairedFlag() && !record.getMateUnmappedFlag()) {
              metrics.READS_ALIGNED_IN_PAIRS++;

              // Check that both ends have mapq > minimum
              final Integer mateMq = record.getIntegerAttribute("MQ");
              if (mateMq == null
                  || mateMq >= MAPPING_QUALITY_THRESOLD
                      && record.getMappingQuality() >= MAPPING_QUALITY_THRESOLD) {
                ++this.chimerasDenominator;

                // With both reads mapped we can see if this pair is chimeric
                if (Math.abs(record.getInferredInsertSize()) > maxInsertSize
                    || !record.getReferenceIndex().equals(record.getMateReferenceIndex())) {
                  ++this.chimeras;
                }
              }
            }
          }
        }
      }
Beispiel #5
0
  @Override
  public int doWork(String[] args) {
    File refFile = null;
    com.github.lindenb.jvarkit.util.cli.GetOpt getopt =
        new com.github.lindenb.jvarkit.util.cli.GetOpt();
    int c;
    while ((c = getopt.getopt(args, "hvL:r:")) != -1) {
      switch (c) {
        case 'h':
          printUsage();
          return 0;
        case 'v':
          System.out.println(getVersion());
          return 0;
        case 'L':
          getLogger().setLevel(java.util.logging.Level.parse(getopt.getOptArg()));
          break;
        case 'r':
          refFile = new File(getopt.getOptArg());
          break;
        case ':':
          System.err.println("Missing argument for option -" + getopt.getOptOpt());
          return -1;
        default:
          System.err.println("Unknown option -" + getopt.getOptOpt());
          return -1;
      }
    }

    if (refFile == null) {
      error("Undefined REF file");
      return -1;
    }
    File bamFile = null;
    if (getopt.getOptInd() + 1 != args.length) {
      info("reading from stdin.");
    } else {
      bamFile = new File(args[getopt.getOptInd()]);
    }

    IndexedFastaSequenceFile indexedFastaSequenceFile = null;
    SAMFileReader samFileReader = null;

    try {
      GenomicSequence genomicSequence = null;
      indexedFastaSequenceFile = new IndexedFastaSequenceFile(refFile);
      SAMFileReader.setDefaultValidationStringency(ValidationStringency.SILENT);
      samFileReader = null;
      if (bamFile == null) {
        samFileReader = new SAMFileReader(System.in);
      } else {
        samFileReader = new SAMFileReader(bamFile);
      }
      XMLOutputFactory xmlfactory = XMLOutputFactory.newInstance();
      XMLStreamWriter w = xmlfactory.createXMLStreamWriter(System.out, "UTF-8");
      w.writeStartDocument("UTF-8", "1.0");
      w.writeStartElement("sam");
      w.writeComment(getProgramCommandLine());
      w.writeAttribute("ref", (bamFile == null ? "stdin" : bamFile.getPath()));
      w.writeAttribute("bam", args[1]);

      SAMRecordIterator iter = samFileReader.iterator();
      while (iter.hasNext()) {
        SAMRecord rec = iter.next();

        final byte readbases[] = rec.getReadBases();
        w.writeStartElement("read");

        w.writeStartElement("name");
        w.writeCharacters(rec.getReadName());
        w.writeEndElement();
        w.writeStartElement("sequence");
        w.writeCharacters(new String(readbases));
        w.writeEndElement();
        w.writeStartElement("flags");
        w.writeAttribute("paired", String.valueOf(rec.getReadPairedFlag()));
        w.writeAttribute(
            "failsVendorQual", String.valueOf(rec.getReadFailsVendorQualityCheckFlag()));
        w.writeAttribute("mapped", String.valueOf(!rec.getReadUnmappedFlag()));
        w.writeAttribute("strand", (rec.getReadNegativeStrandFlag() ? "-" : "+"));

        if (rec.getReadPairedFlag()) {
          w.writeAttribute("mate-mapped", String.valueOf(!rec.getMateUnmappedFlag()));
          w.writeAttribute("mate-strand", (rec.getMateNegativeStrandFlag() ? "-" : "+"));
          w.writeAttribute("proper-pair", String.valueOf(rec.getProperPairFlag()));
        }

        w.writeCharacters(String.valueOf(rec.getFlags()));
        w.writeEndElement();
        if (!rec.getReadUnmappedFlag()) {
          w.writeStartElement("qual");
          w.writeCharacters(String.valueOf(rec.getMappingQuality()));
          w.writeEndElement();

          w.writeStartElement("chrom");
          w.writeAttribute("index", String.valueOf(rec.getReferenceIndex()));
          w.writeCharacters(rec.getReferenceName());
          w.writeEndElement();
          w.writeStartElement("pos");
          w.writeCharacters(String.valueOf(rec.getAlignmentStart()));
          w.writeEndElement();
          w.writeStartElement("cigar");
          w.writeCharacters(rec.getCigarString());
          w.writeEndElement();
        }

        if (!rec.getMateUnmappedFlag()) {
          w.writeStartElement("mate-chrom");
          w.writeAttribute("index", String.valueOf(rec.getMateReferenceIndex()));
          w.writeCharacters(rec.getMateReferenceName());
          w.writeEndElement();
          w.writeStartElement("mate-pos");
          w.writeCharacters(String.valueOf(rec.getMateAlignmentStart()));
          w.writeEndElement();
        }

        if (!rec.getReadUnmappedFlag()) {
          if (genomicSequence == null
              || genomicSequence.getChrom().equals(rec.getReferenceName())) {
            genomicSequence = new GenomicSequence(indexedFastaSequenceFile, rec.getReferenceName());
          }

          w.writeStartElement("align");

          int readIndex = 0;
          int refIndex = rec.getAlignmentStart();

          for (final CigarElement e : rec.getCigar().getCigarElements()) {
            switch (e.getOperator()) {
              case H:
                break; // ignore hard clips
              case P:
                break; // ignore pads
              case I: // cont.
              case S:
                {
                  final int length = e.getLength();
                  for (int i = 0; i < length; ++i) {
                    w.writeEmptyElement(e.getOperator().name());
                    w.writeAttribute("read-index", String.valueOf(readIndex + 1));
                    if (readIndex >= 0 && readIndex < readbases.length) {
                      w.writeAttribute("read-base", String.valueOf((char) (readbases[readIndex])));
                    }
                    readIndex++;
                  }
                  break;
                }
              case N: // cont. -- reference skip
              case D:
                {
                  final int length = e.getLength();
                  for (int i = 0; i < length; ++i) {
                    w.writeEmptyElement(e.getOperator().name());
                    w.writeAttribute("ref-index", String.valueOf(refIndex));
                    if (refIndex >= 1 && refIndex <= genomicSequence.length()) {
                      w.writeAttribute(
                          "ref-base", String.valueOf(genomicSequence.charAt(refIndex - 1)));
                    }
                    refIndex++;
                  }
                  break;
                }
              case M:
              case EQ:
              case X:
                {
                  final int length = e.getLength();
                  for (int i = 0; i < length; ++i) {
                    w.writeEmptyElement(e.getOperator().name());
                    char baseRead = '\0';
                    if (readIndex >= 0 && readIndex < readbases.length) {
                      baseRead = (char) (rec.getReadBases()[readIndex]);
                      w.writeAttribute("read-index", String.valueOf(readIndex + 1));
                      w.writeAttribute("read-base", String.valueOf(baseRead));
                    }
                    w.writeAttribute("ref-index", String.valueOf(refIndex));
                    if (refIndex >= 1 && refIndex <= genomicSequence.length()) {
                      char baseRef = genomicSequence.charAt(refIndex - 1);
                      w.writeAttribute("ref-base", String.valueOf(baseRef));
                      if (Character.toUpperCase(baseRef) != Character.toUpperCase(baseRead)) {
                        w.writeAttribute("mismatch", "true");
                      }
                    }

                    refIndex++;
                    readIndex++;
                  }
                  break;
                }

              default:
                throw new IllegalStateException(
                    "Case statement didn't deal with cigar op: " + e.getOperator());
            }
          }
        }

        w.writeEndElement();

        w.writeEndElement();

        iter.close();
        w.writeEndElement();
      }
      w.writeEndElement();
      w.writeEndDocument();
      w.flush();
      w.close();
    } catch (Exception err) {
      error(err);
      return -1;
    } finally {
      CloserUtil.close(samFileReader);
      CloserUtil.close(indexedFastaSequenceFile);
    }
    return 0;
  }
Beispiel #6
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();
    }
  }