Beispiel #1
0
  /**
   * Finds the adaptor boundary around the read and returns the first base inside the adaptor that
   * is closest to the read boundary. If the read is in the positive strand, this is the first base
   * after the end of the fragment (Picard calls it 'insert'), if the read is in the negative
   * strand, this is the first base before the beginning of the fragment.
   *
   * <p>There are two cases we need to treat here:
   *
   * <p>1) Our read is in the reverse strand :
   *
   * <p><----------------------| * |--------------------->
   *
   * <p>in these cases, the adaptor boundary is at the mate start (minus one)
   *
   * <p>2) Our read is in the forward strand :
   *
   * <p>|----------------------> * <----------------------|
   *
   * <p>in these cases the adaptor boundary is at the start of the read plus the inferred insert
   * size (plus one)
   *
   * @param read the read being tested for the adaptor boundary
   * @return the reference coordinate for the adaptor boundary (effectively the first base IN the
   *     adaptor, closest to the read. NULL if the read is unmapped or the mate is mapped to another
   *     contig.
   */
  public static Integer getAdaptorBoundary(final SAMRecord read) {
    final int MAXIMUM_ADAPTOR_LENGTH = 8;
    final int insertSize =
        Math.abs(
            read
                .getInferredInsertSize()); // the inferred insert size can be negative if the mate
                                           // is mapped before the read (so we take the absolute
                                           // value)

    if (insertSize == 0
        || read
            .getReadUnmappedFlag()) // no adaptors in reads with mates in another chromosome or
                                    // unmapped pairs
    return null;

    Integer
        adaptorBoundary; // the reference coordinate for the adaptor boundary (effectively the first
                         // base IN the adaptor, closest to the read)
    if (read.getReadNegativeStrandFlag())
      adaptorBoundary = read.getMateAlignmentStart() - 1; // case 1 (see header)
    else adaptorBoundary = read.getAlignmentStart() + insertSize + 1; // case 2 (see header)

    if ((adaptorBoundary < read.getAlignmentStart() - MAXIMUM_ADAPTOR_LENGTH)
        || (adaptorBoundary > read.getAlignmentEnd() + MAXIMUM_ADAPTOR_LENGTH))
      adaptorBoundary =
          null; // we are being conservative by not allowing the adaptor boundary to go beyond what
                // we belive is the maximum size of an adaptor

    return adaptorBoundary;
  }
      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;
                }
              }
            }
          }
        }
      }