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/*
* Copyright (C) 2015 Google Inc.
*
* Licensed under the Apache License, Version 2.0 (the "License"); you may not
* use this file except in compliance with the License. You may obtain a copy of
* the License at
*
* http://www.apache.org/licenses/LICENSE-2.0
*
* Unless required by applicable law or agreed to in writing, software
* distributed under the License is distributed on an "AS IS" BASIS, WITHOUT
* WARRANTIES OR CONDITIONS OF ANY KIND, either express or implied. See the
* License for the specific language governing permissions and limitations under
* the License.
*/
package com.google.cloud.genomics.dataflow.readers.bam;
import com.google.api.services.storage.Storage;
import com.google.api.services.storage.Storage.Objects;
import com.google.cloud.dataflow.sdk.transforms.DoFn;
import com.google.cloud.genomics.utils.Contig;
import com.google.common.base.Function;
import com.google.common.collect.Iterables;
import com.google.common.collect.Lists;
import com.google.common.collect.Maps;
import htsjdk.samtools.BAMFileIndexImpl;
import htsjdk.samtools.Bin;
import htsjdk.samtools.Chunk;
import htsjdk.samtools.GenomicIndexUtil;
import htsjdk.samtools.SAMFileHeader;
import htsjdk.samtools.SAMSequenceRecord;
import htsjdk.samtools.SamReader;
import htsjdk.samtools.ValidationStringency;
import htsjdk.samtools.seekablestream.SeekableStream;
import htsjdk.samtools.util.IOUtil;
import java.io.IOException;
import java.util.BitSet;
import java.util.HashMap;
import java.util.List;
import java.util.logging.Level;
import java.util.logging.Logger;
import javax.annotation.Nullable;
/**
* Generates shards for a given BAM file and a desired set of contigs.
*/
public class Sharder {
private static final Logger LOG = Logger.getLogger(Sharder.class.getName());
public interface SharderOutput {
public void output(BAMShard shard);
}
Storage.Objects storageClient;
String filePath;
Iterable requestedContigs;
SharderOutput output;
final ShardingPolicy shardingPolicy;
SamReader reader;
SeekableStream indexStream;
SAMFileHeader header;
BAMFileIndexImpl index;
boolean allReferences;
boolean hasIndex;
HashMap contigsByReference;
public static List shardBAMFile(Objects storageClient,
String filePath, Iterable requestedContigs,
ShardingPolicy shardingPolicy) throws IOException {
final List shards = Lists.newArrayList();
(new Sharder(storageClient, filePath, requestedContigs, shardingPolicy,
new SharderOutput() {
@Override
public void output(BAMShard shard) {
shards.add(shard);
}
}))
.process();
return shards;
}
public Sharder(Objects storageClient, String filePath, Iterable requestedContigs,
ShardingPolicy shardingPolicy,
SharderOutput output) {
super();
this.storageClient = storageClient;
this.filePath = filePath;
this.shardingPolicy = shardingPolicy;
// HTSJDK expects 1-based coordinates - need to shift by 1
this.requestedContigs = Iterables.transform(requestedContigs,
new Function() {
@Override
@Nullable
public Contig apply(@Nullable Contig arg0) {
return new Contig(arg0.referenceName, arg0.start + 1, arg0.end);
}
});
this.output = output;
}
public void process() throws IOException {
LOG.info("Processing BAM file " + filePath);
openFile();
processHeader();
for (SAMSequenceRecord sequenceRecord : header.getSequenceDictionary().getSequences()) {
final Contig contig = desiredContigForReference(sequenceRecord);
if (contig == null) {
continue;
} else if (!hasIndex) {
// No index, so create a shard over all BAM file and pass a contig restrict.
// We will iterate over all reads for this reference and skip the ones outside contig.
LOG.info("No index: outputting shard for " + contig);
output.output(new BAMShard(filePath, null, contig));
continue;
}
createShardsForReference(sequenceRecord, contig);
}
if (index != null) {
index.close();
}
}
void openFile() throws IOException {
final BAMIO.ReaderAndIndex r = BAMIO.openBAMAndExposeIndex(storageClient, filePath, ValidationStringency.DEFAULT_STRINGENCY);
reader = r.reader;
indexStream = r.index;
header = reader.getFileHeader();
hasIndex = reader.hasIndex() && reader.indexing().hasBrowseableIndex();
LOG.info("Has index = " + hasIndex);
if (hasIndex) {
index = new BAMFileIndexImpl(
IOUtil.maybeBufferedSeekableStream(indexStream),header.getSequenceDictionary());
} else {
index = null;
}
}
void processHeader() {
contigsByReference = Maps.newHashMap();
for (Contig contig : requestedContigs) {
contigsByReference.put(contig.referenceName != null ? contig.referenceName : "", contig);
}
if (contigsByReference.size() == 0 || contigsByReference.containsKey("*")) {
LOG.info("Outputting unmapped reads shard ");
output.output(new BAMShard(filePath, null, new Contig("*", 0, -1)));
}
allReferences =
contigsByReference.size() == 0 || contigsByReference.containsKey("");
LOG.info("All references = " + allReferences);
LOG.info("BAM has index = " + reader.hasIndex());
LOG.info("BAM has browseable index = " + reader.indexing().hasBrowseableIndex());
LOG.info("Class for index = " + reader.indexing().getIndex().getClass().getName());
}
Contig desiredContigForReference(SAMSequenceRecord reference) {
Contig contig = contigsByReference.get(reference.getSequenceName());
if (contig == null) {
if (allReferences) {
contig = new Contig(reference.getSequenceName(), 0, -1);
} else {
LOG.fine("No contig requested for reference " + reference.getSequenceName());
return null;
}
}
assert contig != null;
if (contig.end <= 0) {
contig = new Contig(contig.referenceName, contig.start, reference.getSequenceLength());
LOG.info("Updated length for contig for " + contig.referenceName + " to " + contig.end);
}
return contig;
}
void createShardsForReference(SAMSequenceRecord reference, Contig contig) {
LOG.info("Creating shard for: " + contig);
final BitSet overlappingBins = GenomicIndexUtil.regionToBins(
(int) contig.start, (int) contig.end);
if (overlappingBins == null) {
LOG.warning("No overlapping bins for " + contig.start + ":" + contig.end);
return;
}
BAMShard currentShard = null;
for (int binIndex = overlappingBins.nextSetBit(0); binIndex >= 0; binIndex = overlappingBins.nextSetBit(binIndex + 1)) {
final Bin bin = index.getBinData(reference.getSequenceIndex(), binIndex);
if (bin == null) {
continue;
}
if (LOG.isLoggable(Level.FINE)) {
LOG.fine("Processing bin " + index.getFirstLocusInBin(bin) + "-"
+ index.getLastLocusInBin(bin));
}
if (index.getLevelForBin(bin) != (GenomicIndexUtil.LEVEL_STARTS.length - 1)) {
if (LOG.isLoggable(Level.FINEST)) {
LOG.finest("Skipping - not lowest");
}
continue;
// Skip non-lowest level bins
// Its ok to skip higher level bins
// because in BAMShard#finalize we
// will get all chunks overlapping a genomic region that
// we end up having for the shard, so we will not
// miss any reads on the boundary.
}
final List chunksInBin = bin.getChunkList();
if (chunksInBin.isEmpty()) {
if (LOG.isLoggable(Level.FINEST)) {
LOG.finest("Skipping - empty");
}
continue; // Skip empty bins
}
if (currentShard == null) {
int startLocus =
Math.max(index.getFirstLocusInBin(bin), (int)contig.start);
if (LOG.isLoggable(Level.FINE)) {
LOG.fine("Creating shard starting from " + startLocus);
}
currentShard = new BAMShard(filePath, reference.getSequenceName(),
startLocus);
}
currentShard.addBin(chunksInBin, index.getLastLocusInBin(bin));
if (LOG.isLoggable(Level.FINE)) {
LOG.fine("Extending the shard to " + index.getLastLocusInBin(bin));
}
if (shardingPolicy.shardBigEnough(currentShard)) {
LOG.info("Shard size is big enough to finalize: " +
currentShard.sizeInLoci() + ", " + currentShard.approximateSizeInBytes() + " bytes");
final BAMShard bamShard = currentShard.finalize(index, Math.min(index.getLastLocusInBin(bin), (int)contig.end));
LOG.info("Outputting shard: " + bamShard.contig);
output.output(bamShard);
currentShard = null;
}
}
if (currentShard != null) {
LOG.info("Outputting last shard of size " +
currentShard.sizeInLoci() + ", " + currentShard.approximateSizeInBytes() + " bytes "
+ currentShard.contig);
output.output(currentShard.finalize(index, (int)contig.end));
}
}
}
