/**
* 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 int compareCoordinates(final SAMRecord record1, final SAMRecord record2) {
final int seqIndex1 = record1.getReferenceIndex();
final int seqIndex2 = record2.getReferenceIndex();
if (seqIndex1 == -1) {
return ((seqIndex2 == -1) ? 0 : -1);
} else if (seqIndex2 == -1) {
return 1;
}
int result = seqIndex1 - seqIndex2;
if (result != 0) {
return result;
}
result = record1.getAlignmentStart() - record2.getAlignmentStart();
return result;
}
/**
* HACK TO CREATE GATKSAMRECORD BASED ONLY A SAMRECORD FOR TESTING PURPOSES ONLY
*
* @param read
*/
public GATKSAMRecord(final SAMRecord read) {
super(read.getHeader());
super.setReferenceIndex(read.getReferenceIndex());
super.setAlignmentStart(read.getAlignmentStart());
super.setReadName(read.getReadName());
super.setMappingQuality(read.getMappingQuality());
// indexing bin done below
super.setCigar(read.getCigar());
super.setFlags(read.getFlags());
super.setMateReferenceIndex(read.getMateReferenceIndex());
super.setMateAlignmentStart(read.getMateAlignmentStart());
super.setInferredInsertSize(read.getInferredInsertSize());
SAMReadGroupRecord samRG = read.getReadGroup();
SAMBinaryTagAndValue samAttr = GATKBin.getReadBinaryAttributes(read);
if (samAttr == null) {
clearAttributes();
} else {
setAttributes(samAttr);
}
if (samRG != null) {
GATKSAMReadGroupRecord rg = new GATKSAMReadGroupRecord(samRG);
setReadGroup(rg);
}
super.setFileSource(read.getFileSource());
super.setReadName(read.getReadName());
super.setCigarString(read.getCigarString());
super.setReadBases(read.getReadBases());
super.setBaseQualities(read.getBaseQualities());
// From SAMRecord constructor: Do this after the above because setCigarString will clear it.
GATKBin.setReadIndexingBin(this, GATKBin.getReadIndexingBin(read));
}
/** Return the sort key used for the given sort order. Useful in error messages. */
public String getSortKey(final SAMRecord rec) {
switch (sortOrder) {
case coordinate:
return rec.getReferenceName() + ":" + rec.getAlignmentStart();
case queryname:
return rec.getReadName();
case unsorted:
default:
return null;
}
}
/**
* Record any index information for a given BAM record
*
* @param rec The BAM record. Requires rec.getFileSource() is non-null.
*/
public void processAlignment(final SAMRecord rec) {
// metadata
indexStats.recordMetaData(rec);
if (rec.getAlignmentStart() == SAMRecord.NO_ALIGNMENT_START) {
return; // do nothing for records without coordinates, but count them
}
// various checks
final int reference = rec.getReferenceIndex();
if (reference != currentReference) {
throw new SAMException(
"Unexpected reference "
+ reference
+ " when constructing index for "
+ currentReference
+ " for record "
+ rec);
}
binningIndexBuilder.processFeature(
new BinningIndexBuilder.FeatureToBeIndexed() {
@Override
public int getStart() {
return rec.getAlignmentStart();
}
@Override
public int getEnd() {
return rec.getAlignmentEnd();
}
@Override
public Integer getIndexingBin() {
final Integer binNumber = rec.getIndexingBin();
return (binNumber == null ? rec.computeIndexingBin() : binNumber);
}
@Override
public Chunk getChunk() {
final SAMFileSource source = rec.getFileSource();
if (source == null) {
throw new SAMException(
"No source (virtual file offsets); needed for indexing on BAM Record " + rec);
}
return ((BAMFileSpan) source.getFilePointer()).getSingleChunk();
}
});
}
private boolean passesFilter(
final SAMRecord record,
final String sequence,
final int startPos,
final int endPos,
final boolean contained) {
if (record == null) {
return false;
}
if (!safeEquals(record.getReferenceName(), sequence)) {
return false;
}
final int alignmentStart = record.getAlignmentStart();
int alignmentEnd = record.getAlignmentEnd();
if (alignmentStart <= 0) {
assertTrue(record.getReadUnmappedFlag());
return false;
}
if (alignmentEnd <= 0) {
// For indexing-only records, treat as single base alignment.
assertTrue(record.getReadUnmappedFlag());
alignmentEnd = alignmentStart;
}
if (contained) {
if (startPos != 0 && alignmentStart < startPos) {
return false;
}
if (endPos != 0 && alignmentEnd > endPos) {
return false;
}
} else {
if (startPos != 0 && alignmentEnd < startPos) {
return false;
}
if (endPos != 0 && alignmentStart > endPos) {
return false;
}
}
return true;
}
public void find_coverage(SAMResource sres) {
int start_base = sres.region.range.start;
int end_base = sres.region.range.end;
int coverage_len = (end_base - start_base) + 1;
int i, end, ref_i, read_i, len;
int[] coverage = new int[coverage_len];
Arrays.fill(coverage, 0);
WorkingFile wf = null;
if (outfile != null) {
try {
wf = new WorkingFile(outfile);
ps = wf.getPrintStream();
} catch (Exception e) {
System.err.println("I/O error: " + e); // debug
e.printStackTrace();
System.exit(1);
}
}
try {
//
// gather coverage info:
//
CloseableIterator<SAMRecord> iterator = sres.get_iterator();
int read_count = 0;
int ref_min = -1;
int ref_max = -1;
while (iterator.hasNext()) {
SAMRecord sr = iterator.next();
read_count++;
// System.err.println(sr.getReadName() + ": " + sr.getAlignmentStart() + "-" +
// sr.getAlignmentEnd()); // debug
if (sr.getReadUnmappedFlag()) continue;
if (sr.getDuplicateReadFlag()) {
if (verbose_mode)
System.err.println(
sr.getReadName()
+ "."
+ (sr.getReadNegativeStrandFlag() ? "R" : "F")
+ " ignoring, duplicate");
continue;
}
byte[] read = sr.getReadBases();
byte[] quals = sr.getBaseQualities();
for (AlignmentBlock ab : sr.getAlignmentBlocks()) {
len = ab.getLength();
read_i = ab.getReadStart() - 1;
ref_i = ab.getReferenceStart() - start_base;
if (ref_min == -1 || ref_i < ref_min) ref_min = ref_i;
for (i = read_i, end = read_i + len; i < end; i++, ref_i++) {
if (ref_i >= 0 && ref_i < coverage_len) {
if (quals[i] >= MIN_QUALITY) {
if (verbose_mode)
System.err.println(
sr.getReadName()
+ "."
+ (sr.getReadNegativeStrandFlag() ? "R" : "F")
+ " hit at "
+ (ref_i + start_base)
+ " as="
+ sr.getAlignmentStart()
+ " ae="
+ sr.getAlignmentEnd());
coverage[ref_i]++;
} else if (verbose_mode) {
System.err.println(
sr.getReadName()
+ "."
+ (sr.getReadNegativeStrandFlag() ? "R" : "F")
+ " qual_reject at "
+ (ref_i + start_base)
+ " as="
+ sr.getAlignmentStart()
+ " ae="
+ sr.getAlignmentEnd());
}
}
}
if (ref_max == -1 || ref_i > ref_max) ref_max = ref_i;
}
}
sres.close();
System.err.println(
"records:"
+ read_count
+ " ref_min:"
+ (ref_min + start_base)
+ " ref_max:"
+ (ref_max + start_base)); // debug
//
// report coverage info:
//
for (i = 0; i < coverage.length; i++) {
if (name != null) ps.print(name + ",");
ps.println((i + start_base) + "," + coverage[i]); // debug
}
if (wf != null) wf.finish();
} catch (Exception e) {
System.err.println("ERROR: " + e); // debug
e.printStackTrace();
}
}
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);
}
@Override
public void execute() {
log.info("Initializing kmer code map...");
Map<Character, Integer> kmerCodeIndices = new HashMap<Character, Integer>();
kmerCodeIndices.put('0', 1);
kmerCodeIndices.put('A', 3);
kmerCodeIndices.put('B', 4);
kmerCodeIndices.put('C', 5);
kmerCodeIndices.put('_', 6);
kmerCodeIndices.put('.', 7);
kmerCodeIndices.put('1', 9);
Map<Character, String> kmerCodeNames = new LinkedHashMap<Character, String>();
kmerCodeNames.put('0', "ref0");
kmerCodeNames.put('A', "repetitive");
kmerCodeNames.put('B', "both");
kmerCodeNames.put('C', "lowcoverage");
kmerCodeNames.put('_', "lowconfidence");
kmerCodeNames.put('.', "novel");
kmerCodeNames.put('1', "ref1");
if (KMER_CODE_NAMES != null) {
for (Character c : kmerCodeNames.keySet()) {
String cStr = String.valueOf(c);
if (KMER_CODE_NAMES.containsKey(cStr)) {
kmerCodeNames.put(c, KMER_CODE_NAMES.get(cStr));
}
}
}
for (Character c : kmerCodeNames.keySet()) {
log.info(" {} {}: {}", c, kmerCodeIndices.get(c), kmerCodeNames.get(c));
}
log.info("Loading annotated contigs...");
Map<String, Map<String, String>> annotatedContigs = new HashMap<String, Map<String, String>>();
int kmerSize = 0;
if (ANN.length() > 0) {
TableReader tr = new TableReader(ANN);
for (Map<String, String> te : tr) {
String contigName = te.get("contigName");
if (kmerSize == 0) {
kmerSize = te.get("seq").length() - te.get("kmerOrigin").length() + 1;
}
annotatedContigs.put(contigName, te);
String[] ref0ToCanonicalExact =
(te.get("ref0ToCanonicalExact").equals("NA")
|| te.get("ref0ToCanonicalExact").equals("*:0-0")
? "NA:0-0"
: te.get("ref0ToCanonicalExact"))
.split("[:-]");
String[] ref1ToCanonicalExact =
(te.get("ref1ToCanonicalExact").equals("NA")
|| te.get("ref1ToCanonicalExact").equals("*:0-0")
? "NA:0-0"
: te.get("ref1ToCanonicalExact"))
.split("[:-]");
cout.println(
te.get("sampleName")
+ "_"
+ te.get("accession")
+ "_"
+ contigName
+ " "
+ ref0ToCanonicalExact[0]
+ " "
+ ref0ToCanonicalExact[1]
+ " "
+ ref0ToCanonicalExact[2]
+ " radius1=0.8r");
cout.println(
te.get("sampleName")
+ "_"
+ te.get("accession")
+ "_"
+ contigName
+ " "
+ ref1ToCanonicalExact[0]
+ " "
+ ref1ToCanonicalExact[1]
+ " "
+ ref1ToCanonicalExact[2]
+ " radius2=0.6r");
}
}
log.info(" contigs: {}", annotatedContigs.size());
log.info(" kmer size: {}", kmerSize);
log.info("Computing kmer inheritance information...");
SAMFileHeader sfh = CONTIGS.getFileHeader();
for (Character c : kmerCodeNames.keySet()) {
SAMReadGroupRecord rgr = new SAMReadGroupRecord(kmerCodeNames.get(c));
rgr.setSample(kmerCodeNames.get(c));
sfh.addReadGroup(rgr);
}
SAMFileWriterFactory sfwf = new SAMFileWriterFactory();
sfwf.setCreateIndex(true);
SAMFileWriter sfw = sfwf.makeBAMWriter(sfh, false, bout);
TableWriter tw = new TableWriter(sout);
Set<IGVEntry> igvEntries = new TreeSet<IGVEntry>();
int numContigs = 0;
for (SAMRecord contig : CONTIGS) {
if (CONTIG_NAMES == null
|| CONTIG_NAMES.isEmpty()
|| CONTIG_NAMES.contains(contig.getReadName())) {
Map<String, String> te = annotatedContigs.get(contig.getReadName());
if (annotatedContigs.containsKey(contig.getReadName())) {
String seq = contig.getReadString();
// log.debug(" te: {}", te);
String annSeq = te.get("seq");
String kmerOrigin = te.get("kmerOrigin");
Map<CortexKmer, Character> kmerCodes = new HashMap<CortexKmer, Character>();
for (int i = 0; i < kmerOrigin.length(); i++) {
CortexKmer kmer = new CortexKmer(annSeq.substring(i, i + kmerSize));
Character code = kmerOrigin.charAt(i);
kmerCodes.put(kmer, code);
}
Map<Character, Integer> kmerStats = new HashMap<Character, Integer>();
for (Character c : kmerCodeNames.keySet()) {
kmerStats.put(c, 0);
}
boolean changed = false;
// We want to be able to examine soft-clipped regions as well.
List<CigarElement> ces = new ArrayList<CigarElement>();
for (CigarElement ce : contig.getCigar().getCigarElements()) {
if (ce.getOperator().equals(CigarOperator.S)) {
ces.add(new CigarElement(ce.getLength(), CigarOperator.M));
changed = true;
} else {
ces.add(ce);
}
}
if (changed) {
CigarElement firstCe = contig.getCigar().getCigarElements().get(0);
if (firstCe.getOperator().equals(CigarOperator.S)) {
contig.setAlignmentStart(contig.getAlignmentStart() - firstCe.getLength());
}
contig.setCigar(new Cigar(ces));
}
for (AlignmentBlock ab : contig.getAlignmentBlocks()) {
for (int i = ab.getReadStart() - 1; i < ab.getReadStart() + ab.getLength(); i++) {
if (i + kmerSize < seq.length()) {
CortexKmer kmer = new CortexKmer(seq.substring(i, i + kmerSize));
SAMRecord skmer = new SAMRecord(CONTIGS.getFileHeader());
skmer.setReadBases(seq.substring(i, i + kmerSize).getBytes());
List<CigarElement> cigarElements = new ArrayList<CigarElement>();
cigarElements.add(new CigarElement(kmerSize, CigarOperator.M));
Cigar cigar = new Cigar(cigarElements);
skmer.setReadName(contig.getReadName() + "." + kmer.getKmerAsString());
skmer.setReferenceName(contig.getReferenceName());
skmer.setCigar(cigar);
skmer.setReadPairedFlag(false);
skmer.setDuplicateReadFlag(false);
skmer.setMateNegativeStrandFlag(false);
skmer.setAlignmentStart(ab.getReferenceStart() - ab.getReadStart() + 1 + i);
skmer.setAttribute("RG", "none");
skmer.setMappingQuality(0);
Character c = kmerCodes.get(kmer);
String codeName = kmerCodeNames.get(c);
String parentReadGroupId = null;
String sampleReadGroupId = null;
for (SAMReadGroupRecord rgr : sfh.getReadGroups()) {
if (rgr.getSample().equals(codeName)) {
parentReadGroupId = rgr.getReadGroupId();
}
if (rgr.getSample().equals(contig.getReadGroup().getSample())) {
sampleReadGroupId = rgr.getReadGroupId();
}
}
skmer.setAttribute(
"RG", parentReadGroupId != null ? parentReadGroupId : sampleReadGroupId);
skmer.setMappingQuality(99);
sfw.addAlignment(skmer);
kmerStats.put(c, kmerStats.get(c) + 1);
IGVEntry igvEntry = new IGVEntry();
igvEntry.chromosome = contig.getReferenceName();
igvEntry.start = ab.getReferenceStart() - ab.getReadStart() + i;
igvEntry.parentageName = kmerCodeNames.get(c);
igvEntry.parentage = kmerCodeIndices.get(c);
igvEntries.add(igvEntry);
}
}
}
if (!contig.isSecondaryOrSupplementary()) {
beout.println(
contig.getReferenceName()
+ "\t"
+ contig.getAlignmentStart()
+ "\t"
+ contig.getAlignmentEnd()
+ "\t"
+ contig.getReadName()
+ "."
+ contig.getReadGroup().getSample());
if (annotatedContigs.size() > 10 && numContigs % (annotatedContigs.size() / 10) == 0) {
log.info(" processed {}/{} contigs", numContigs, annotatedContigs.size());
}
numContigs++;
}
Map<String, String> stats = new LinkedHashMap<String, String>();
stats.put("contigName", contig.getReadName());
stats.put("sampleName", contig.getReadGroup().getSample());
for (Character c : kmerCodeNames.keySet()) {
stats.put(kmerCodeNames.get(c), String.valueOf(kmerStats.get(c)));
}
tw.addEntry(stats);
}
}
}
log.info("Writing kmer inheritance information...");
out.printf("%s\t%s\t%s\t%s\t%s\n", "Chromosome", "Start", "End", "Feature", "Parentage");
for (IGVEntry igvEntry : igvEntries) {
out.printf(
"%s\t%d\t%d\t%s\t%d\n",
igvEntry.chromosome,
igvEntry.start,
igvEntry.start + 1,
igvEntry.parentageName,
igvEntry.parentage);
}
sfw.close();
}
