htsjdk.samtools.SAMUtils Maven / Gradle / Ivy
/*
* The MIT License
*
* Copyright (c) 2009 The Broad Institute
*
* Permission is hereby granted, free of charge, to any person obtaining a copy
* of this software and associated documentation files (the "Software"), to deal
* in the Software without restriction, including without limitation the rights
* to use, copy, modify, merge, publish, distribute, sublicense, and/or sell
* copies of the Software, and to permit persons to whom the Software is
* furnished to do so, subject to the following conditions:
*
* The above copyright notice and this permission notice shall be included in
* all copies or substantial portions of the Software.
*
* THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR
* IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY,
* FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE
* AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER
* LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM,
* OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN
* THE SOFTWARE.
*/
package htsjdk.samtools;
import htsjdk.samtools.seekablestream.SeekableStream;
import htsjdk.samtools.util.BinaryCodec;
import htsjdk.samtools.util.CigarUtil;
import htsjdk.samtools.util.CloserUtil;
import htsjdk.samtools.util.CoordMath;
import htsjdk.samtools.util.RuntimeEOFException;
import htsjdk.samtools.util.StringUtil;
import htsjdk.tribble.annotation.Strand;
import java.io.File;
import java.io.IOException;
import java.io.UnsupportedEncodingException;
import java.math.BigInteger;
import java.security.MessageDigest;
import java.security.NoSuchAlgorithmException;
import java.util.ArrayList;
import java.util.Collections;
import java.util.List;
import java.util.Locale;
import java.util.Map;
import java.util.Optional;
import java.util.TreeMap;
import java.util.regex.Pattern;
/**
* Utilty methods.
*/
public final class SAMUtils {
/**
* regex for semicolon, used in {@link SAMUtils#getOtherCanonicalAlignments(SAMRecord)}
*/
private static final Pattern SEMICOLON_PAT = Pattern.compile("[;]");
/**
* regex for comma, used in {@link SAMUtils#getOtherCanonicalAlignments(SAMRecord)}
*/
private static final Pattern COMMA_PAT = Pattern.compile("[,]");
// Representation of bases, one for when in low-order nybble, one for when in high-order nybble.
private static final byte COMPRESSED_EQUAL_LOW = 0;
private static final byte COMPRESSED_A_LOW = 1;
private static final byte COMPRESSED_C_LOW = 2;
private static final byte COMPRESSED_M_LOW = 3;
private static final byte COMPRESSED_G_LOW = 4;
private static final byte COMPRESSED_R_LOW = 5;
private static final byte COMPRESSED_S_LOW = 6;
private static final byte COMPRESSED_V_LOW = 7;
private static final byte COMPRESSED_T_LOW = 8;
private static final byte COMPRESSED_W_LOW = 9;
private static final byte COMPRESSED_Y_LOW = 10;
private static final byte COMPRESSED_H_LOW = 11;
private static final byte COMPRESSED_K_LOW = 12;
private static final byte COMPRESSED_D_LOW = 13;
private static final byte COMPRESSED_B_LOW = 14;
private static final byte COMPRESSED_N_LOW = 15;
private static final byte COMPRESSED_EQUAL_HIGH = COMPRESSED_EQUAL_LOW << 4;
private static final byte COMPRESSED_A_HIGH = COMPRESSED_A_LOW << 4;
private static final byte COMPRESSED_C_HIGH = COMPRESSED_C_LOW << 4;
private static final byte COMPRESSED_G_HIGH = COMPRESSED_G_LOW << 4;
private static final byte COMPRESSED_T_HIGH = (byte) (COMPRESSED_T_LOW << 4);
private static final byte COMPRESSED_N_HIGH = (byte) (COMPRESSED_N_LOW << 4);
private static final byte COMPRESSED_M_HIGH = (byte) (COMPRESSED_M_LOW << 4);
private static final byte COMPRESSED_R_HIGH = (byte) (COMPRESSED_R_LOW << 4);
private static final byte COMPRESSED_S_HIGH = (byte) (COMPRESSED_S_LOW << 4);
private static final byte COMPRESSED_V_HIGH = (byte) (COMPRESSED_V_LOW << 4);
private static final byte COMPRESSED_W_HIGH = (byte) (COMPRESSED_W_LOW << 4);
private static final byte COMPRESSED_Y_HIGH = (byte) (COMPRESSED_Y_LOW << 4);
private static final byte COMPRESSED_H_HIGH = (byte) (COMPRESSED_H_LOW << 4);
private static final byte COMPRESSED_K_HIGH = (byte) (COMPRESSED_K_LOW << 4);
private static final byte COMPRESSED_D_HIGH = (byte) (COMPRESSED_D_LOW << 4);
private static final byte COMPRESSED_B_HIGH = (byte) (COMPRESSED_B_LOW << 4);
private static final byte[] COMPRESSED_LOOKUP_TABLE = {
'=',
'A',
'C',
'M',
'G',
'R',
'S',
'V',
'T',
'W',
'Y',
'H',
'K',
'D',
'B',
'N'
};
public static final int MAX_PHRED_SCORE = 93;
/**
* Convert from a byte array containing =AaCcGgTtNnMmRrSsVvWwYyHhKkDdBb represented as ASCII, to a byte array half as long,
* with for example, =, A, C, G, T converted to 0, 1, 2, 4, 8, 15.
*
* @param readBases Bases as ASCII bytes.
* @return New byte array with bases represented as nybbles, in BAM binary format.
*/
static byte[] bytesToCompressedBases(final byte[] readBases) {
final byte[] compressedBases = new byte[(readBases.length + 1) / 2];
int i;
for (i = 1; i < readBases.length; i += 2) {
compressedBases[i / 2] = (byte) (charToCompressedBaseHigh(readBases[i - 1]) |
charToCompressedBaseLow(readBases[i]));
}
// Last nybble
if (i == readBases.length) {
compressedBases[i / 2] = charToCompressedBaseHigh(readBases[i - 1]);
}
return compressedBases;
}
/**
* Convert from a byte array with bases stored in nybbles, with for example,=, A, C, G, T, N represented as 0, 1, 2, 4, 8, 15,
* to a a byte array containing =AaCcGgTtNn represented as ASCII.
*
* @param length Number of bases (not bytes) to convert.
* @param compressedBases Bases represented as nybbles, in BAM binary format.
* @param compressedOffset Byte offset in compressedBases to start.
* @return New byte array with bases as ASCII bytes.
*/
public static byte[] compressedBasesToBytes(final int length, final byte[] compressedBases, final int compressedOffset) {
final byte[] ret = new byte[length];
int i;
for (i = 1; i < length; i += 2) {
final int compressedIndex = i / 2 + compressedOffset;
ret[i - 1] = compressedBaseToByteHigh(compressedBases[compressedIndex]);
ret[i] = compressedBaseToByteLow(compressedBases[compressedIndex]);
}
// Last nybble
if (i == length) {
ret[i - 1] = compressedBaseToByteHigh(compressedBases[i / 2 + compressedOffset]);
}
return ret;
}
/**
* Convert from ASCII byte to BAM nybble representation of a base in low-order nybble.
*
* @param base One of =AaCcGgTtNnMmRrSsVvWwYyHhKkDdBb.
* @return Low-order nybble-encoded equivalent.
* @throws IllegalArgumentException if the base is not one of =AaCcGgTtNnMmRrSsVvWwYyHhKkDdBb.
*/
private static byte charToCompressedBaseLow(final byte base) {
switch (base) {
case '=':
return COMPRESSED_EQUAL_LOW;
case 'a':
case 'A':
return COMPRESSED_A_LOW;
case 'c':
case 'C':
return COMPRESSED_C_LOW;
case 'g':
case 'G':
return COMPRESSED_G_LOW;
case 't':
case 'T':
return COMPRESSED_T_LOW;
case 'n':
case 'N':
case '.':
return COMPRESSED_N_LOW;
// IUPAC ambiguity codes
case 'M':
case 'm':
return COMPRESSED_M_LOW;
case 'R':
case 'r':
return COMPRESSED_R_LOW;
case 'S':
case 's':
return COMPRESSED_S_LOW;
case 'V':
case 'v':
return COMPRESSED_V_LOW;
case 'W':
case 'w':
return COMPRESSED_W_LOW;
case 'Y':
case 'y':
return COMPRESSED_Y_LOW;
case 'H':
case 'h':
return COMPRESSED_H_LOW;
case 'K':
case 'k':
return COMPRESSED_K_LOW;
case 'D':
case 'd':
return COMPRESSED_D_LOW;
case 'B':
case 'b':
return COMPRESSED_B_LOW;
default:
throw new IllegalArgumentException("Bad base passed to charToCompressedBaseLow: " + Character.toString((char) base) + "(" + base + ")");
}
}
/**
* Convert from ASCII byte to BAM nybble representation of a base in high-order nybble.
*
* @param base One of =AaCcGgTtNnMmRrSsVvWwYyHhKkDdBb.
* @return High-order nybble-encoded equivalent.
* @throws IllegalArgumentException if the base is not one of =AaCcGgTtNnMmRrSsVvWwYyHhKkDdBb.
*/
private static byte charToCompressedBaseHigh(final byte base) {
switch (base) {
case '=':
return COMPRESSED_EQUAL_HIGH;
case 'a':
case 'A':
return COMPRESSED_A_HIGH;
case 'c':
case 'C':
return COMPRESSED_C_HIGH;
case 'g':
case 'G':
return COMPRESSED_G_HIGH;
case 't':
case 'T':
return COMPRESSED_T_HIGH;
case 'n':
case 'N':
case '.':
return COMPRESSED_N_HIGH;
// IUPAC ambiguity codes
case 'M':
case 'm':
return COMPRESSED_M_HIGH;
case 'R':
case 'r':
return COMPRESSED_R_HIGH;
case 'S':
case 's':
return COMPRESSED_S_HIGH;
case 'V':
case 'v':
return COMPRESSED_V_HIGH;
case 'W':
case 'w':
return COMPRESSED_W_HIGH;
case 'Y':
case 'y':
return COMPRESSED_Y_HIGH;
case 'H':
case 'h':
return COMPRESSED_H_HIGH;
case 'K':
case 'k':
return COMPRESSED_K_HIGH;
case 'D':
case 'd':
return COMPRESSED_D_HIGH;
case 'B':
case 'b':
return COMPRESSED_B_HIGH;
default:
throw new IllegalArgumentException("Bad base passed to charToCompressedBaseHigh: " + Character.toString((char) base) + "(" + base + ")");
}
}
/**
* Returns the byte corresponding to a certain nybble
*
* @param base One of COMPRESSED_*_LOW, a low-order nybble encoded base.
* @return ASCII base, one of =ACGTNMRSVWYHKDB.
* @throws IllegalArgumentException if the base is not one of =ACGTNMRSVWYHKDB.
*/
private static byte compressedBaseToByte(byte base) {
try {
return COMPRESSED_LOOKUP_TABLE[base];
} catch (IndexOutOfBoundsException e) {
throw new IllegalArgumentException("Bad base passed to charToCompressedBase: " + Character.toString((char) base) + "(" + base + ")");
}
}
/**
* Convert from BAM nybble representation of a base in low-order nybble to ASCII byte.
*
* @param base One of COMPRESSED_*_LOW, a low-order nybble encoded base.
* @return ASCII base, one of ACGTN=.
*/
private static byte compressedBaseToByteLow(final int base) {
return compressedBaseToByte((byte) (base & 0xf));
}
/**
* Convert from BAM nybble representation of a base in high-order nybble to ASCII byte.
*
* @param base One of COMPRESSED_*_HIGH, a high-order nybble encoded base.
* @return ASCII base, one of ACGTN=.
*/
private static byte compressedBaseToByteHigh(final int base) {
return compressedBaseToByte((byte) ((base >> 4) & 0xf));
}
/**
* Convert bases in place into canonical form, upper case, and with no-call represented as N.
*
* @param bases byte array of bases to "normalize", in place.
*/
static void normalizeBases(final byte[] bases) {
for (int i = 0; i < bases.length; ++i) {
bases[i] = StringUtil.toUpperCase(bases[i]);
if (bases[i] == '.') {
bases[i] = 'N';
}
}
}
/**
* Convert an array of bytes, in which each byte is a binary phred quality score, to
* printable ASCII representation of the quality scores, ala FASTQ format.
*
* Equivalent to phredToFastq(data, 0, data.length)
*
* @param data Array of bytes in which each byte is a binar phred score.
* @return String with ASCII representation of those quality scores.
*/
public static String phredToFastq(final byte[] data) {
if (data == null) {
return null;
}
return phredToFastq(data, 0, data.length);
}
/**
* Convert an array of bytes, in which each byte is a binary phred quality score, to
* printable ASCII representation of the quality scores, ala FASTQ format.
*
* @param buffer Array of bytes in which each byte is a binar phred score.
* @param offset Where in buffer to start conversion.
* @param length How many bytes of buffer to convert.
* @return String with ASCII representation of those quality scores.
*/
public static String phredToFastq(final byte[] buffer, final int offset, final int length) {
final char[] chars = new char[length];
for (int i = 0; i < length; i++) {
chars[i] = phredToFastq(buffer[offset + i] & 0xFF);
}
return new String(chars);
}
/**
* Convert a single binary phred score to printable ASCII representation, ala FASTQ format.
*
* @param phredScore binary phred score.
* @return Printable ASCII representation of phred score.
*/
public static char phredToFastq(final int phredScore) {
if (phredScore < 0 || phredScore > MAX_PHRED_SCORE) {
throw new IllegalArgumentException("Cannot encode phred score: " + phredScore);
}
return (char) (33 + phredScore);
}
/**
* Convert a string with phred scores in printable ASCII FASTQ format to an array
* of binary phred scores.
*
* @param fastq Phred scores in FASTQ printable ASCII format.
* @return byte array of binary phred scores in which each byte corresponds to a character in the input string.
*/
public static byte[] fastqToPhred(final String fastq) {
if (fastq == null) {
return null;
}
final int length = fastq.length();
final byte[] scores = new byte[length];
for (int i = 0; i < length; i++) {
scores[i] = (byte) fastqToPhred(fastq.charAt(i));
}
return scores;
}
/**
* Converts printable qualities in Sanger fastq format to binary phred scores.
*/
public static void fastqToPhred(final byte[] fastq) {
for (int i = 0; i < fastq.length; ++i) {
fastq[i] = (byte) fastqToPhred((char) (fastq[i] & 0xff));
}
}
/**
* Convert a single printable ASCII FASTQ format phred score to binary phred score.
*
* @param ch Printable ASCII FASTQ format phred score.
* @return Binary phred score.
*/
public static int fastqToPhred(final char ch) {
if (ch < 33 || ch > 126) {
throw new IllegalArgumentException("Invalid fastq character: " + ch);
}
return (ch - 33);
}
/**
* calculate the bin given an alignment in [beg,end)
* Copied from SAM spec.
*
* @param beg 0-based start of read (inclusive)
* @param end 0-based end of read (exclusive)
* @deprecated Use {@link GenomicIndexUtil#regionToBin}
*/
@Deprecated
static int reg2bin(final int beg, final int end) {
return GenomicIndexUtil.regionToBin(beg, end);
}
/**
* Handle a list of validation errors according to the validation stringency.
*
* @param validationErrors List of errors to report, or null if there are no errors.
* @param samRecordIndex Record number of the SAMRecord corresponding to the validation errors, or -1 if
* the record number is not known.
* @param validationStringency If STRICT, throw a SAMFormatException. If LENIENT, print the validation
* errors to stderr. If SILENT, do nothing.
*/
public static void processValidationErrors(final List validationErrors,
final long samRecordIndex,
final ValidationStringency validationStringency) {
if (validationErrors != null && !validationErrors.isEmpty()) {
for (final SAMValidationError validationError : validationErrors) {
validationError.setRecordNumber(samRecordIndex);
}
if (validationStringency == ValidationStringency.STRICT) {
throw new SAMFormatException("SAM validation error: " + validationErrors.get(0));
} else if (validationStringency == ValidationStringency.LENIENT) {
for (final SAMValidationError error : validationErrors) {
System.err.println("Ignoring SAM validation error: " + error);
}
}
}
}
public static void processValidationError(final SAMValidationError validationError,
final ValidationStringency validationStringency) {
if (validationStringency == ValidationStringency.STRICT) {
throw new SAMFormatException("SAM validation error: " + validationError);
} else if (validationStringency == ValidationStringency.LENIENT) {
System.err.println("Ignoring SAM validation error: " + validationError);
}
}
private static final SAMHeaderRecordComparator HEADER_RECORD_COMPARATOR =
new SAMHeaderRecordComparator<>(
SAMReadGroupRecord.PLATFORM_UNIT_TAG,
SAMReadGroupRecord.LIBRARY_TAG,
SAMReadGroupRecord.DATE_RUN_PRODUCED_TAG,
SAMReadGroupRecord.READ_GROUP_SAMPLE_TAG,
SAMReadGroupRecord.SEQUENCING_CENTER_TAG,
SAMReadGroupRecord.PLATFORM_TAG,
SAMReadGroupRecord.DESCRIPTION_TAG,
SAMReadGroupRecord.READ_GROUP_ID_TAG
// We don't actually want to compare with ID but it's suitable
// "just in case" since it's the only one that's actually required
);
/**
* Calculate a hash code from identifying information in the RG (read group) records in a SAM file's
* header. This hash code changes any time read groups are added or removed. Comparing one file's
* hash code to another's tells you if the read groups in the BAM files are different.
*/
public static String calculateReadGroupRecordChecksum(final File input, final File referenceFasta) {
final String ENCODING = "UTF-8";
final MessageDigest digest;
try {
digest = MessageDigest.getInstance("MD5");
} catch (final NoSuchAlgorithmException nsae) {
throw new Error("No MD5 algorithm was available in a Java JDK? Unheard-of!");
}
// Sort the read group records by their first
final SamReader reader = SamReaderFactory.makeDefault().referenceSequence(referenceFasta).open(input);
final List sortedRecords = new ArrayList<>(reader.getFileHeader().getReadGroups());
Collections.sort(sortedRecords, HEADER_RECORD_COMPARATOR);
for (final SAMReadGroupRecord rgRecord : sortedRecords) {
final TreeMap sortedAttributes = new TreeMap<>();
for (final Map.Entry attributeEntry : rgRecord.getAttributes()) {
sortedAttributes.put(attributeEntry.getKey(), attributeEntry.getValue());
}
try {
for (final Map.Entry sortedEntry : sortedAttributes.entrySet()) {
if (!sortedEntry.getKey().equals(SAMReadGroupRecord.READ_GROUP_ID_TAG)) { // Redundant check, safety first
digest.update(sortedEntry.getKey().getBytes(ENCODING));
digest.update(sortedEntry.getValue().getBytes(ENCODING));
}
}
} catch (final UnsupportedEncodingException uee) {
throw new Error("No " + ENCODING + "!? WTH?");
}
}
// Convert to a String and pad to get the full 32 chars.
final StringBuilder hashText = new StringBuilder((new BigInteger(1, digest.digest())).toString(16));
while (hashText.length() < 32) hashText.insert(0, "0");
CloserUtil.close(reader);
return hashText.toString();
}
/**
* Chains program in front of the first "head" item in the list of
* SAMProgramRecords in header. This method should not be used
* when there are multiple chains of program groups in a header, only when
* it can safely be assumed that there is only one chain. It correctly handles
* the case where program has already been added to the header, so
* it can be used whether creating a SAMProgramRecord with a constructor or when
* calling SAMFileHeader.createProgramRecord().
*/
public static void chainSAMProgramRecord(final SAMFileHeader header, final SAMProgramRecord program) {
final List pgs = header.getProgramRecords();
if (!pgs.isEmpty()) {
final List referencedIds = new ArrayList<>();
for (final SAMProgramRecord pg : pgs) {
if (pg.getPreviousProgramGroupId() != null) {
referencedIds.add(pg.getPreviousProgramGroupId());
}
}
for (final SAMProgramRecord pg : pgs) {
// if record being chained has already been added, ignore it
if (pg.getProgramGroupId().equals(program.getProgramGroupId())) {
continue;
}
if (!referencedIds.contains(pg.getProgramGroupId())) {
program.setPreviousProgramGroupId(pg.getProgramGroupId());
break;
}
}
}
}
/**
* Strip mapping information from a SAMRecord.
*
* WARNING: by clearing the secondary and supplementary flags,
* this may have the affect of producing multiple distinct records with the
* same read name and flags, which may lead to invalid SAM/BAM output.
* Callers of this method should make sure to deal with this issue.
*/
public static void makeReadUnmapped(final SAMRecord rec) {
if (rec.getReadNegativeStrandFlag()) {
rec.reverseComplement(true);
rec.setReadNegativeStrandFlag(false);
}
rec.setDuplicateReadFlag(false);
rec.setReferenceIndex(SAMRecord.NO_ALIGNMENT_REFERENCE_INDEX);
rec.setAlignmentStart(SAMRecord.NO_ALIGNMENT_START);
rec.setCigarString(SAMRecord.NO_ALIGNMENT_CIGAR);
rec.setMappingQuality(SAMRecord.NO_MAPPING_QUALITY);
rec.setInferredInsertSize(0);
rec.setSecondaryAlignment(false);
rec.setSupplementaryAlignmentFlag(false);
rec.setProperPairFlag(false);
rec.setReadUnmappedFlag(true);
}
/**
* Strip mapping information from a SAMRecord, but preserve it in the 'O' tags if it isn't already set.
*/
public static void makeReadUnmappedWithOriginalTags(final SAMRecord rec) {
if (!hasOriginalMappingInformation(rec)) {
rec.setAttribute(SAMTag.OP, rec.getAlignmentStart());
rec.setAttribute(SAMTag.OC, rec.getCigarString());
rec.setAttribute(SAMTag.OF, rec.getFlags());
rec.setAttribute(SAMTag.OR, rec.getReferenceName());
}
makeReadUnmapped(rec);
}
/**
* See if any tags pertaining to original mapping information have been set.
*/
public static boolean hasOriginalMappingInformation(final SAMRecord rec) {
return rec.getAttribute(SAMTag.OP) != null
|| rec.getAttribute(SAMTag.OC) != null
|| rec.getAttribute(SAMTag.OF) != null
|| rec.getAttribute(SAMTag.OR) != null;
}
/**
* Determines if a cigar has any element that both consumes read bases and consumes reference bases
* (e.g. is not all soft-clipped)
*/
public static boolean cigarMapsNoBasesToRef(final Cigar cigar) {
for (final CigarElement el : cigar.getCigarElements()) {
if (el.getOperator().consumesReadBases() && el.getOperator().consumesReferenceBases()) {
return false;
}
}
return true;
}
/**
* Tests if the provided record is mapped entirely beyond the end of the reference (i.e., the alignment start is greater than the
* length of the sequence to which the record is mapped).
*
* @param record must not have a null SamFileHeader
*/
public static boolean recordMapsEntirelyBeyondEndOfReference(final SAMRecord record) {
if (record.getHeader() == null) {
throw new SAMException("A non-null SAMHeader is required to resolve the mapping position: " + record.getReadName());
} else {
return record.getHeader().getSequence(record.getReferenceIndex()).getSequenceLength() < record.getAlignmentStart();
}
}
/**
* @return negative if mapq1 < mapq2, etc.
* Note that MAPQ(0) < MAPQ(255) < MAPQ(1)
*/
public static int compareMapqs(final int mapq1, final int mapq2) {
if (mapq1 == mapq2) return 0;
if (mapq1 == 0) return -1;
else if (mapq2 == 0) return 1;
else if (mapq1 == 255) return -1;
else if (mapq2 == 255) return 1;
else return mapq1 - mapq2;
}
/**
* Hokey algorithm for combining two MAPQs into values that are comparable, being cognizant of the fact
* that in MAPQ world, 1 > 255 > 0. In this algorithm, 255 is treated as if it were 0.01, so that
* CombinedMapq(1,0) > CombinedMapq(255, 255) > CombinedMapq(0, 0).
* The return value should not be used for anything other than comparing to the return value of other
* invocations of this method.
*/
public static int combineMapqs(int m1, int m2) {
if (m1 == 255) {
m1 = 1;
} else {
m1 *= 100;
}
if (m2 == 255) {
m2 = 1;
} else {
m2 *= 100;
}
return m1 + m2;
}
/**
* Returns the virtual file offset of the first record in a BAM file - i.e. the virtual file
* offset after skipping over the text header and the sequence records.
*/
public static long findVirtualOffsetOfFirstRecordInBam(final File bamFile) {
try {
return BAMFileReader.findVirtualOffsetOfFirstRecord(bamFile);
} catch (final IOException ioe) {
throw new RuntimeEOFException(ioe);
}
}
/**
* Returns the virtual file offset of the first record in a BAM file - i.e. the virtual file
* offset after skipping over the text header and the sequence records.
*/
public static long findVirtualOffsetOfFirstRecordInBam(final SeekableStream seekableStream) {
try {
return BAMFileReader.findVirtualOffsetOfFirstRecord(seekableStream);
} catch (final IOException ioe) {
throw new RuntimeEOFException(ioe);
}
}
/**
* Given a Cigar, Returns blocks of the sequence that have been aligned directly to the
* reference sequence. Note that clipped portions, and inserted and deleted bases (vs. the reference)
* are not represented in the alignment blocks.
*
* @param cigar The cigar containing the alignment information
* @param alignmentStart The start (1-based) of the alignment
* @param cigarTypeName The type of cigar passed - for error logging.
* @return List of alignment blocks
*/
public static List getAlignmentBlocks(final Cigar cigar, final int alignmentStart, final String cigarTypeName) {
if (cigar == null) return Collections.emptyList();
final List alignmentBlocks = new ArrayList<>();
int readBase = 1;
int refBase = alignmentStart;
for (final CigarElement e : cigar.getCigarElements()) {
switch (e.getOperator()) {
case H:
break; // ignore hard clips
case P:
break; // ignore pads
case S:
readBase += e.getLength();
break; // soft clip read bases
case N:
refBase += e.getLength();
break; // reference skip
case D:
refBase += e.getLength();
break;
case I:
readBase += e.getLength();
break;
case M:
case EQ:
case X:
final int length = e.getLength();
alignmentBlocks.add(new AlignmentBlock(readBase, refBase, length));
readBase += length;
refBase += length;
break;
default:
throw new IllegalStateException("Case statement didn't deal with " + cigarTypeName + " op: " + e.getOperator() + "in CIGAR: " + cigar);
}
}
return Collections.unmodifiableList(alignmentBlocks);
}
/**
* @param alignmentStart The start (1-based) of the alignment
* @param cigar The cigar containing the alignment information
* @return the alignment start (1-based, inclusive) adjusted for clipped bases. For example if the read
* has an alignment start of 100 but the first 4 bases were clipped (hard or soft clipped)
* then this method will return 96.
*
* Invalid to call on an unmapped read.
* Invalid to call with cigar = null
*/
public static int getUnclippedStart(final int alignmentStart, final Cigar cigar) {
int unClippedStart = alignmentStart;
for (final CigarElement cig : cigar.getCigarElements()) {
final CigarOperator op = cig.getOperator();
if (op == CigarOperator.SOFT_CLIP || op == CigarOperator.HARD_CLIP) {
unClippedStart -= cig.getLength();
} else {
break;
}
}
return unClippedStart;
}
/**
* @param alignmentEnd The end (1-based) of the alignment
* @param cigar The cigar containing the alignment information
* @return the alignment end (1-based, inclusive) adjusted for clipped bases. For example if the read
* has an alignment end of 100 but the last 7 bases were clipped (hard or soft clipped)
* then this method will return 107.
*
* Invalid to call on an unmapped read.
* Invalid to call with cigar = null
*/
public static int getUnclippedEnd(final int alignmentEnd, final Cigar cigar) {
int unClippedEnd = alignmentEnd;
final List cigs = cigar.getCigarElements();
for (int i = cigs.size() - 1; i >= 0; --i) {
final CigarElement cig = cigs.get(i);
final CigarOperator op = cig.getOperator();
if (op == CigarOperator.SOFT_CLIP || op == CigarOperator.HARD_CLIP) {
unClippedEnd += cig.getLength();
} else {
break;
}
}
return unClippedEnd;
}
/**
* Returns the Mate Cigar String as stored in the attribute 'MC'.
*
* @param rec the SAM record
* @return Mate Cigar String, or null if there is none.
*/
public static String getMateCigarString(final SAMRecord rec) {
return rec.getStringAttribute(SAMTag.MC);
}
/**
* Returns the Mate Cigar or null if there is none.
*
* @param rec the SAM record
* @param withValidation true if we are to validate the mate cigar before returning, false otherwise.
* @return Cigar object for the read's mate, or null if there is none.
*/
public static Cigar getMateCigar(final SAMRecord rec, final boolean withValidation) {
final String mateCigarString = getMateCigarString(rec);
Cigar mateCigar = null;
if (mateCigarString != null) {
mateCigar = TextCigarCodec.decode(mateCigarString);
if (withValidation && rec.getValidationStringency() != ValidationStringency.SILENT) {
final List alignmentBlocks = getAlignmentBlocks(mateCigar, rec.getMateAlignmentStart(), "mate cigar");
SAMUtils.processValidationErrors(validateCigar(rec, mateCigar, rec.getMateReferenceIndex(), alignmentBlocks, -1, "Mate CIGAR"), -1L, rec.getValidationStringency());
}
}
return mateCigar;
}
/**
* Returns the Mate Cigar or null if there is none. No validation is done on the returned cigar.
*
* @param rec the SAM record
* @return Cigar object for the read's mate, or null if there is none.
*/
public static Cigar getMateCigar(final SAMRecord rec) {
return getMateCigar(rec, false);
}
/**
* @param rec the SAM record
* @return number of cigar elements (number + operator) in the mate cigar string.
*/
public static int getMateCigarLength(final SAMRecord rec) {
final Cigar mateCigar = getMateCigar(rec);
return (mateCigar != null) ? mateCigar.numCigarElements() : 0;
}
/**
* This method uses the MateCigar value as determined from the attribute MC. It must be non-null.
*
* @param rec the SAM record
* @return 1-based inclusive rightmost position of the clipped mate sequence, or 0 read if unmapped.
*/
public static int getMateAlignmentEnd(final SAMRecord rec) {
if (rec.getMateUnmappedFlag()) {
throw new RuntimeException("getMateAlignmentEnd called on an unmapped mate: " + rec);
}
final Cigar mateCigar = SAMUtils.getMateCigar(rec);
if (mateCigar == null) {
throw new SAMException("Mate CIGAR (Tag MC) not found:" + rec);
}
return CoordMath.getEnd(rec.getMateAlignmentStart(), mateCigar.getReferenceLength());
}
/**
* @param rec the SAM record
* @return the mate alignment start (1-based, inclusive) adjusted for clipped bases. For example if the mate
* has an alignment start of 100 but the first 4 bases were clipped (hard or soft clipped)
* then this method will return 96.
*
* Invalid to call on an unmapped read.
*/
public static int getMateUnclippedStart(final SAMRecord rec) {
if (rec.getMateUnmappedFlag())
throw new RuntimeException("getMateUnclippedStart called on an unmapped mate: " + rec);
final Cigar mateCigar = getMateCigar(rec);
if (mateCigar == null) {
throw new SAMException("Mate CIGAR (Tag MC) not found: " + rec);
}
return SAMUtils.getUnclippedStart(rec.getMateAlignmentStart(), mateCigar);
}
/**
* @param rec the SAM record
* @return the mate alignment end (1-based, inclusive) adjusted for clipped bases. For example if the mate
* has an alignment end of 100 but the last 7 bases were clipped (hard or soft clipped)
* then this method will return 107.
*
* Invalid to call on an unmapped read.
*/
public static int getMateUnclippedEnd(final SAMRecord rec) {
if (rec.getMateUnmappedFlag()) {
throw new RuntimeException("getMateUnclippedEnd called on an unmapped mate: " + rec);
}
final Cigar mateCigar = SAMUtils.getMateCigar(rec);
if (mateCigar == null) {
throw new SAMException("Mate CIGAR (Tag MC) not found: " + rec);
}
return SAMUtils.getUnclippedEnd(getMateAlignmentEnd(rec), mateCigar);
}
/**
* @param rec the SAM record
* Returns blocks of the mate sequence that have been aligned directly to the
* reference sequence. Note that clipped portions of the mate and inserted and
* deleted bases (vs. the reference) are not represented in the alignment blocks.
*/
public static List getMateAlignmentBlocks(final SAMRecord rec) {
return getAlignmentBlocks(getMateCigar(rec), rec.getMateAlignmentStart(), "mate cigar");
}
/**
* Run all validations of the mate's CIGAR. These include validation that the CIGAR makes sense independent of
* placement, plus validation that CIGAR + placement yields all bases with M operator within the range of the reference.
*
* @param rec the SAM record
* @param cigar The cigar containing the alignment information
* @param referenceIndex The reference index
* @param alignmentBlocks The alignment blocks (parsed from the cigar)
* @param recordNumber For error reporting. -1 if not known.
* @param cigarTypeName For error reporting. "Read CIGAR" or "Mate Cigar"
* @return List of errors, or null if no errors.
*/
public static List validateCigar(final SAMRecord rec,
final Cigar cigar,
final Integer referenceIndex,
final List alignmentBlocks,
final long recordNumber,
final String cigarTypeName) {
// Don't know line number, and don't want to force read name to be decoded.
List ret = cigar.isValid(rec.getReadName(), recordNumber);
if (referenceIndex != SAMRecord.NO_ALIGNMENT_REFERENCE_INDEX) {
SAMFileHeader samHeader = rec.getHeader();
if (null == samHeader) {
if (ret == null) ret = new ArrayList<>();
ret.add(new SAMValidationError(SAMValidationError.Type.MISSING_HEADER,
cigarTypeName + " A non-null SAMHeader is required to validate cigar elements for: ", rec.getReadName(), recordNumber));
} else {
final SAMSequenceRecord sequence = samHeader.getSequence(referenceIndex);
final int referenceSequenceLength = sequence.getSequenceLength();
for (final AlignmentBlock alignmentBlock : alignmentBlocks) {
if (alignmentBlock.getReferenceStart() + alignmentBlock.getLength() - 1 > referenceSequenceLength) {
if (ret == null) ret = new ArrayList<>();
ret.add(new SAMValidationError(SAMValidationError.Type.CIGAR_MAPS_OFF_REFERENCE,
cigarTypeName + " M operator maps off end of reference", rec.getReadName(), recordNumber));
break;
}
}
}
}
return ret;
}
/**
* Run all validations of the mate's CIGAR. These include validation that the CIGAR makes sense independent of
* placement, plus validation that CIGAR + placement yields all bases with M operator within the range of the reference.
*
* @param rec the SAM record
* @param recordNumber For error reporting. -1 if not known.
* @return List of errors, or null if no errors.
*/
public static List validateMateCigar(final SAMRecord rec, final long recordNumber) {
List ret = null;
if (rec.getValidationStringency() != ValidationStringency.SILENT) {
if (rec.getReadPairedFlag() && !rec.getMateUnmappedFlag()) { // The mateCigar will be defined if the mate is mapped
if (getMateCigarString(rec) != null) {
ret = SAMUtils.validateCigar(rec, getMateCigar(rec), rec.getMateReferenceIndex(), getMateAlignmentBlocks(rec), recordNumber, "Mate CIGAR");
}
} else {
if (getMateCigarString(rec) != null) {
ret = new ArrayList<>();
if (!rec.getReadPairedFlag()) {
// If the read is not paired, and the Mate Cigar String (MC Attribute) exists, that is a validation error
ret.add(new SAMValidationError(SAMValidationError.Type.MATE_CIGAR_STRING_INVALID_PRESENCE,
"Mate CIGAR String (MC Attribute) present for a read that is not paired", rec.getReadName(), recordNumber));
} else { // will hit here if rec.getMateUnmappedFlag() is true
// If the Mate is unmapped, and the Mate Cigar String (MC Attribute) exists, that is a validation error.
ret.add(new SAMValidationError(SAMValidationError.Type.MATE_CIGAR_STRING_INVALID_PRESENCE,
"Mate CIGAR String (MC Attribute) present for a read whose mate is unmapped", rec.getReadName(), recordNumber));
}
}
}
}
return ret;
}
/**
* Checks to see if it is valid for this record to have a mate CIGAR (MC) and then if there is a mate CIGAR available. This is done by
* checking that this record is paired, its mate is mapped, and that it returns a non-null mate CIGAR.
*
* @param rec
* @return
*/
public static boolean hasMateCigar(SAMRecord rec) {
// NB: use getMateCigarString rather than getMateCigar to avoid validation.
return (rec.getReadPairedFlag() && !rec.getMateUnmappedFlag() && null != SAMUtils.getMateCigarString(rec));
}
/**
* Returns a string that is the the read group ID and read name separated by a colon. This is meant to canonically
* identify a given record within a set of records.
*
* @param record SAMRecord for which "canonical" read name is requested
* @return The record's readgroup-id (if non-null) and the read name, separated by a colon, ':'
*/
public static String getCanonicalRecordName(final SAMRecord record) {
String name = record.getStringAttribute(ReservedTagConstants.READ_GROUP_ID);
if (null == name) name = record.getReadName();
else name = name + ":" + record.getReadName();
return name;
}
/**
* Returns the number of bases that need to be clipped due to overlapping pairs. If the record is not paired,
* or the given record's start position is greater than its mate's start position, zero is automatically returned.
* NB: This method assumes that the record's mate is not contained within the given record's alignment.
*
* @param rec SAMRecord that needs clipping due to overlapping pairs.
* @return the number of bases at the end of the read that need to be clipped such that there would be no overlapping bases with its mate.
* Read bases include only those from insertion, match, or mismatch Cigar operators.
*/
public static int getNumOverlappingAlignedBasesToClip(final SAMRecord rec) {
// NB: ignores how to handle supplemental records when present for both ends by just using the mate information in the record.
if (!rec.getReadPairedFlag() || rec.getReadUnmappedFlag() || rec.getMateUnmappedFlag()) return 0;
// Only clip records that are left-most in genomic order and overlapping.
if (rec.getMateAlignmentStart() < rec.getAlignmentStart()) return 0; // right-most, so ignore.
else if (rec.getMateAlignmentStart() == rec.getAlignmentStart() && rec.getFirstOfPairFlag())
return 0; // same start, so pick the first end
// Find the number of read bases after the given mate's alignment start.
int numBasesToClip = 0;
final int refStartPos = rec.getMateAlignmentStart(); // relative reference position after which we should start clipping
final Cigar cigar = rec.getCigar();
int refPos = rec.getAlignmentStart();
for (final CigarElement el : cigar.getCigarElements()) {
final CigarOperator operator = el.getOperator();
final int refBasesLength = operator.consumesReferenceBases() ? el.getLength() : 0;
if (refStartPos <= refPos + refBasesLength - 1) { // add to clipped bases
if (operator == CigarOperator.MATCH_OR_MISMATCH) { // M
if (refStartPos < refPos) numBasesToClip += refBasesLength; // use all of the bases
else
numBasesToClip += (refPos + refBasesLength) - refStartPos; // since the mate's alignment start can be in the middle of a cigar element
} else if (operator == CigarOperator.SOFT_CLIP || operator == CigarOperator.HARD_CLIP || operator == CigarOperator.PADDING || operator == CigarOperator.SKIPPED_REGION) {
// ignore
} else { // ID
numBasesToClip += operator.consumesReadBases() ? el.getLength() : 0; // clip all the bases in the read from this operator
}
}
refPos += refBasesLength;
}
if (numBasesToClip < 0) return 0; // left-most but not overlapping
return numBasesToClip;
}
/**
* Returns a (possibly new) record that has been clipped if input is a mapped paired and has overlapping bases with its mate.
* See {@link #getNumOverlappingAlignedBasesToClip(SAMRecord)} for how the number of overlapping bases is computed.
* NB: this does not properly consider a cigar like: 100M20S10H.
* NB: This method assumes that the record's mate is not contained within the given record's alignment.
*
* @param record the record from which to clip bases.
* @param noSideEffects if true a modified clone of the original record is returned, otherwise we modify the record directly.
* @return a (possibly new) record that has been clipped
*/
public static SAMRecord clipOverlappingAlignedBases(final SAMRecord record, final boolean noSideEffects) {
return clipOverlappingAlignedBases(record, getNumOverlappingAlignedBasesToClip(record), noSideEffects);
}
/**
* Returns a (possibly new) SAMRecord with the given number of bases soft-clipped at the end of the read if is a mapped
* paired and has overlapping bases with its mate.
* NB: this does not properly consider a cigar like: 100M20S10H.
* NB: This method assumes that the record's mate is not contained within the given record's alignment.
*
* @param record the record from which to clip bases.
* @param numOverlappingBasesToClip the number of bases to clip at the end of the read.
* @param noSideEffects if true a modified clone of the original record is returned, otherwise we modify the record directly.
* @return Returns a (possibly new) SAMRecord with the given number of bases soft-clipped
*/
public static SAMRecord clipOverlappingAlignedBases(final SAMRecord record, final int numOverlappingBasesToClip, final boolean noSideEffects) {
// NB: ignores how to handle supplemental records when present for both ends by just using the mate information in the record.
if (numOverlappingBasesToClip <= 0 || record.getReadUnmappedFlag() || record.getMateUnmappedFlag()) {
return record;
}
try {
final SAMRecord rec = noSideEffects ? ((SAMRecord) record.clone()) : record;
// watch out for when the second read overlaps all of the first read
if (rec.getMateAlignmentStart() <= rec.getAlignmentStart()) { // make it unmapped
rec.setReadUnmappedFlag(true);
return rec;
}
// 1-based index of first base in read to clip.
int clipFrom = rec.getReadLength() - numOverlappingBasesToClip + 1;
// we have to check if the last cigar element is soft-clipping, so we can subtract that from clipFrom
final CigarElement cigarElement = rec.getCigar().getCigarElement(rec.getCigarLength() - 1);
if (CigarOperator.SOFT_CLIP == cigarElement.getOperator()) clipFrom -= cigarElement.getLength();
// FIXME: does not properly consider a cigar like: 100M20S10H
// clip it, clip it good
rec.setCigar(new Cigar(CigarUtil.softClipEndOfRead(clipFrom, rec.getCigar().getCigarElements())));
return rec;
} catch (final CloneNotSupportedException e) {
throw new SAMException(e.getMessage(), e);
}
}
/**
* Checks if a long attribute value is within the allowed range of a 32-bit unsigned integer.
*
* @param value a long value to check
* @return true if value is >= 0 and <= {@link BinaryCodec#MAX_UINT}, and false otherwise
*/
public static boolean isValidUnsignedIntegerAttribute(long value) {
return value >= 0 && value <= BinaryCodec.MAX_UINT;
}
/**
* Extract a List of 'other canonical alignments' from a SAM record. Those alignments are stored as a string in the 'SA' tag as defined
* in the SAM specification.
* The name, sequence and qualities, mate data are copied from the original record.
*
* @param record must be non null and must have a non-null associated header.
* @return a list of 'other canonical alignments' SAMRecords. The list is empty if the 'SA' attribute is missing.
*/
public static List getOtherCanonicalAlignments(final SAMRecord record) {
if (record == null) throw new IllegalArgumentException("record is null");
if (record.getHeader() == null) throw new IllegalArgumentException("record.getHeader() is null");
/* extract value of SA tag */
final Object saValue = record.getAttribute(SAMTag.SA);
if (saValue == null) return Collections.emptyList();
if (!(saValue instanceof String)) throw new SAMException(
"Expected a String for attribute 'SA' but got " + saValue.getClass() + ". Record: " + record);
final SAMRecordFactory samReaderFactory = new DefaultSAMRecordFactory();
/* the spec says: "Other canonical alignments in a chimeric alignment, formatted as a
* semicolon-delimited list: (rname,pos,strand,CIGAR,mapQ,NM;)+.
* Each element in the list represents a part of the chimeric alignment.
* Conventionally, at a supplementary line, the 1st element points to the primary line.
*/
/* break string using semicolon */
final String semiColonStrs[] = SEMICOLON_PAT.split((String) saValue);
/* the result list */
final List alignments = new ArrayList<>(semiColonStrs.length);
/* base SAM flag */
int record_flag = record.getFlags();
record_flag &= ~SAMFlag.PROPER_PAIR.flag;
record_flag &= ~SAMFlag.SUPPLEMENTARY_ALIGNMENT.flag;
record_flag &= ~SAMFlag.READ_REVERSE_STRAND.flag;
for (int i = 0; i < semiColonStrs.length; ++i) {
final String semiColonStr = semiColonStrs[i];
/* ignore empty string */
if (semiColonStr.isEmpty()) continue;
/* break string using comma */
final String commaStrs[] = COMMA_PAT.split(semiColonStr);
if (commaStrs.length != 6)
throw new SAMException("Bad 'SA' attribute in " + semiColonStr + ". Record: " + record);
/* create the new record */
final SAMRecord otherRec = samReaderFactory.createSAMRecord(record.getHeader());
/* copy fields from the original record */
otherRec.setReadName(record.getReadName());
otherRec.setReadBases(record.getReadBases());
otherRec.setBaseQualities(record.getBaseQualities());
if (record.getReadPairedFlag() && !record.getMateUnmappedFlag()) {
otherRec.setMateReferenceIndex(record.getMateReferenceIndex());
otherRec.setMateAlignmentStart(record.getMateAlignmentStart());
}
/* get reference sequence */
final int tid = record.getHeader().getSequenceIndex(commaStrs[0]);
if (tid == -1)
throw new SAMException("Unknown contig in " + semiColonStr + ". Record: " + record);
otherRec.setReferenceIndex(tid);
/* fill POS */
final int alignStart;
try {
alignStart = Integer.parseInt(commaStrs[1]);
} catch (final NumberFormatException err) {
throw new SAMException("bad POS in " + semiColonStr + ". Record: " + record, err);
}
otherRec.setAlignmentStart(alignStart);
/* set TLEN */
if (record.getReadPairedFlag() &&
!record.getMateUnmappedFlag() &&
record.getMateReferenceIndex() == tid) {
otherRec.setInferredInsertSize(record.getMateAlignmentStart() - alignStart);
}
/* set FLAG */
int other_flag = record_flag;
other_flag |= (commaStrs[2].equals("+") ? 0 : SAMFlag.READ_REVERSE_STRAND.flag);
/* spec: Conventionally, at a supplementary line, the 1st element points to the primary line */
if (!(record.getSupplementaryAlignmentFlag() && i == 0)) {
other_flag |= SAMFlag.SUPPLEMENTARY_ALIGNMENT.flag;
}
otherRec.setFlags(other_flag);
/* set CIGAR */
otherRec.setCigar(TextCigarCodec.decode(commaStrs[3]));
/* set MAPQ */
try {
otherRec.setMappingQuality(Integer.parseInt(commaStrs[4]));
} catch (final NumberFormatException err) {
throw new SAMException("bad MAPQ in " + semiColonStr + ". Record: " + record, err);
}
/* fill NM */
try {
if (!commaStrs[5].equals("*")) {
otherRec.setAttribute(SAMTag.NM, Integer.parseInt(commaStrs[5]));
}
} catch (final NumberFormatException err) {
throw new SAMException("bad NM in " + semiColonStr + ". Record: " + record, err);
}
/* if strand is not the same: reverse-complement */
if (otherRec.getReadNegativeStrandFlag() != record.getReadNegativeStrandFlag()) {
otherRec.reverseComplement(true);
}
/* add the alignment */
alignments.add(otherRec);
}
return alignments;
}
/**
* @deprecated because the method does the exact opposite of what it says. Use the correctly named
* isReferenceSequenceIncompatibleWithBAI() instead.
*/
@Deprecated public static boolean isReferenceSequenceCompatibleWithBAI(final SAMSequenceRecord sequence) {
return isReferenceSequenceIncompatibleWithBAI(sequence);
}
/**
* Checks if reference sequence is compatible with BAI indexing format.
* @param sequence reference sequence.
*/
public static boolean isReferenceSequenceIncompatibleWithBAI(final SAMSequenceRecord sequence) {
return sequence.getSequenceLength() > GenomicIndexUtil.BIN_GENOMIC_SPAN;
}
/**
* Function to create the OA tag value from a record. The OA tag contains the mapping information
* of a record encoded as a comma-separated string (REF,POS,STRAND,CIGAR,MAPPING_QUALITY,NM_TAG_VALUE)
* @param record to use for generating the OA tag
* @return the OA tag string value
*/
public static String calculateOATagValue(SAMRecord record) {
if (record.getReferenceName().contains(",")) {
throw new SAMException(String.format("Reference name for record %s contains a comma character.", record.getReadName()));
}
final String oaValue;
if (record.getReadUnmappedFlag()) {
oaValue = String.format(Locale.US, "*,0,%s,*,%s,",
record.getReadNegativeStrandFlag() ? Strand.NEGATIVE : Strand.POSITIVE,
record.getMappingQuality());
} else {
oaValue = String.format(Locale.US, "%s,%s,%s,%s,%s,%s",
record.getReferenceName(),
record.getAlignmentStart(),
record.getReadNegativeStrandFlag() ? Strand.NEGATIVE : Strand.POSITIVE,
record.getCigarString(),
record.getMappingQuality(),
Optional.ofNullable(record.getAttribute(SAMTag.NM)).orElse(""));
}
return oaValue;
}
}