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/*++
Module Name:
SAM.cpp
Abstract:
Sequence Alignment Map (SAM) file writer and reader.
Environment:
User mode service.
SamWriter and SamReader (and their subclasses) aren't thread safe.
--*/
#include "stdafx.h"
#include "BigAlloc.h"
#include "Compat.h"
#include "Read.h"
#include "SAM.h"
#include "Bam.h"
#include "Tables.h"
#include "RangeSplitter.h"
#include "ParallelTask.h"
#include "Util.h"
#include "ReadSupplierQueue.h"
#include "FileFormat.h"
#include "AlignerOptions.h"
#include "directions.h"
#include "exit.h"
using std::max;
using std::min;
using util::strnchr;
bool readIdsMatch(const char* id0, const char* id1)
{
for (unsigned i = 0; ; i++) {
char c0 = id0[i];
char c1 = id1[i];
if (c0 != c1) return false;
// don't parse the read ID after the first space or slash, which can represent metadata (or which half of the mate pair the read is).
if (c0 == 0 || c0 == ' ' || c0 == '/') return true;
}
return true;
}
bool readIdsMatch(Read *read0, Read *read1)
{
if (read0->getIdLength() != read1->getIdLength()) {
return false;
}
for (unsigned i = 0; i < read0->getIdLength(); i++) {
char c0 = read0->getId()[i];
char c1 = read1->getId()[i];
if (c0 != c1) return false;
// don't parse the read ID after the first space or slash, which can represent metadata (or which half of the mate pair the read is).
if (c0 == ' ' || c0 == '/') return true;
}
return true;
}
char *
strnchrs(char *str, char charToFind, char charToFind2, size_t maxLen) // Hokey version that looks for either of two chars
{
for (size_t i = 0; i < maxLen; i++) {
if (str[i] == charToFind || str[i] == charToFind2) {
return str + i;
}
if (str[i] == 0) {
return NULL;
}
}
return NULL;
}
char *
SAMReader::skipToBeyondNextFieldSeparator(char *str, const char *endOfBuffer, size_t *o_charsUntilFirstSeparator)
{
if (NULL == str) return NULL;
char *nextChar = str;
while (nextChar < endOfBuffer && *nextChar != '\n' && *nextChar != '\t' && *nextChar != '\r' /* for Windows CRLF text */) {
nextChar++;
}
if (NULL != o_charsUntilFirstSeparator) {
*o_charsUntilFirstSeparator = nextChar - str;
}
if (nextChar >= endOfBuffer || *nextChar == '\n') {
return NULL;
}
while (nextChar < endOfBuffer && ('\t' == *nextChar || '\r' == *nextChar)) {
nextChar++;
}
if (nextChar >= endOfBuffer) {
return NULL;
}
return nextChar;
}
SAMReader *
SAMReader::create(
DataSupplier* supplier,
const char *fileName,
int bufferCount,
const ReaderContext& context,
_int64 startingOffset,
_int64 amountOfFileToProcess)
{
DataReader* data = supplier->getDataReader(bufferCount, maxLineLen, 0.0, 0);
SAMReader *reader = new SAMReader(data, context);
reader->init(fileName, startingOffset, amountOfFileToProcess);
return reader;
}
void
SAMReader::readHeader(const char *fileName)
{
// todo: allow for larger headers
_int64 headerSize = 512 * 1024; // 1M header initially (it's doubled before we use it)
_int64 oldHeaderSize = 0;
char* buffer;
bool sawWholeHeader;
do {
headerSize *= 2;
buffer = data->readHeader(&headerSize);
if (oldHeaderSize >= headerSize) {
//
// No new data, we hit EOF
//
return;
}
oldHeaderSize = headerSize;
if (!parseHeader(fileName, buffer, buffer + headerSize, context.genome, &headerSize, &context.headerMatchesIndex, &sawWholeHeader)) {
WriteErrorMessage("SAMReader: failed to parse header on '%s'\n", fileName);
soft_exit(1);
}
} while (!sawWholeHeader);
_ASSERT(context.header == NULL);
char* p = new char[headerSize + 1];
memcpy(p, buffer, headerSize);
p[headerSize] = 0;
context.header = p;
context.headerBytes = context.headerLength = headerSize;
}
SAMReader::SAMReader(
DataReader* i_data,
const ReaderContext& i_context)
: ReadReader(i_context), data(i_data), headerSize(-1), clipping(i_context.clipping)
{
}
//
// Implement the ReadReader form of getNextRead, which doesn't include the
// alignment results by simply throwing them away.
//
bool
SAMReader::getNextRead(Read *readToUpdate)
{
return getNextRead(readToUpdate, NULL, NULL, NULL, NULL, NULL, NULL);
}
bool
SAMReader::parseHeader(
const char *fileName,
char *firstLine,
char *endOfBuffer,
const Genome *genome,
_int64 *o_headerSize,
bool *o_headerMatchesIndex,
bool *o_sawWholeHeader)
{
char *nextLineToProcess = firstLine;
*o_headerMatchesIndex = true;
int numSQLines = 0;
while (NULL != nextLineToProcess && nextLineToProcess < endOfBuffer && '@' == *nextLineToProcess) {
//
// Make sure we have the complete line.
//
bool foundCompleteLine = false;
for (char *c = nextLineToProcess; c < endOfBuffer; c++) {
if (*c == '\n') {
foundCompleteLine = true;
break;
}
}
if (!foundCompleteLine) {
*o_sawWholeHeader = false;
return true; // Parsed OK, but incomplete
}
if (!strncmp("@SQ",nextLineToProcess,3)) {
//
// These lines represent sequences in the reference genome, what are
// called "contigs" in the Genome class. (Roughly, chromosomes or major
// variants like some versions of the MHC genes on chr6; or more
// particularly the things that come in different FASTA files from the
// reference assembly).
//
// Verify that they actually match what's in our reference genome.
//
numSQLines++;
if (nextLineToProcess + 3 >= endOfBuffer || ' ' != nextLineToProcess[3] && '\t' != nextLineToProcess[3]) {
WriteErrorMessage("Malformed SAM file '%s' has @SQ without a following space or tab.\n",fileName);
return false;
}
char *snStart = nextLineToProcess + 4;
while (snStart < endOfBuffer && strncmp(snStart,"SN:",__min(3,endOfBuffer-snStart)) && *snStart != '\n' && *snStart != 0) {
snStart++;
}
if (snStart >= endOfBuffer || *snStart == '\n' || *snStart == 0) {
WriteErrorMessage("Malformed @SQ line doesn't have 'SN:' in file '%s'\n",fileName);
return false;
}
const size_t contigNameBufferSize = 512;
char contigName[contigNameBufferSize];
for (unsigned i = 0; i < contigNameBufferSize && snStart+3+i < endOfBuffer; i++) {
if (snStart[3+i] == ' ' || snStart[3+i] == '\t' || snStart[3+i] == '\n' || snStart[3+i] == 0) {
contigName[i] = '\0';
} else {
contigName[i] = snStart[3+i];
}
}
contigName[contigNameBufferSize - 1] = '\0';
if (genome == NULL || !genome->getLocationOfContig(contigName, NULL)) {
*o_headerMatchesIndex = false;
}
} else if (!strncmp("@HD",nextLineToProcess,3) || !strncmp("@RG",nextLineToProcess,3) || !strncmp("@PG",nextLineToProcess,3) ||
!strncmp("@CO",nextLineToProcess,3)) {
//
// Ignore these lines.
//
} else {
WriteErrorMessage("Unrecognized header line in SAM file.\n");
return false;
}
char * p = strnchr(nextLineToProcess,'\n',endOfBuffer-nextLineToProcess);
if (p == NULL) {
// no newline, look for null to truncate buffer
p = (char*) memchr(nextLineToProcess, 0, endOfBuffer - nextLineToProcess);
nextLineToProcess = p != NULL ? p + 1 : endOfBuffer;
break;
}
nextLineToProcess = p + 1;
}
*o_headerMatchesIndex &= genome != NULL && numSQLines == genome->getNumContigs();
*o_headerSize = nextLineToProcess - firstLine;
if (NULL != o_sawWholeHeader) {
*o_sawWholeHeader = nextLineToProcess < endOfBuffer;
}
return true;
}
bool
SAMReader::parseLine(char *line, char *endOfBuffer, char *result[], size_t *linelength, size_t fieldLengths[])
{
*linelength = 0;
char *next = line;
char *endOfLine = strnchr(line,'\n',endOfBuffer-line);
if (NULL == endOfLine) {
return false;
}
//
// Skip over any leading spaces and tabs
//
while (next < endOfLine && (*next == ' ' || *next == '\t')) {
next++;
}
for (unsigned i = 0; i < nSAMFields; i++) {
if (NULL == next || next >= endOfLine) {
if (i == OPT) {
// no optional fields
result[OPT] = NULL;
break;
} else {
//
// Too few fields.
//
return false;
}
}
result[i] = next;
if (i == OPT) {
// OPT field is actually all fields until end of line
fieldLengths[OPT] = endOfLine - next;
break;
}
next = skipToBeyondNextFieldSeparator(next,endOfLine,&fieldLengths[i]);
}
*linelength = endOfLine - line + 1; // +1 skips over the \n
return true;
}
void
SAMReader::getReadFromLine(
const Genome *genome,
char *line,
char *endOfBuffer,
Read *read,
AlignmentResult *alignmentResult,
GenomeLocation *out_genomeLocation,
Direction *direction,
unsigned *mapQ,
size_t *lineLength,
unsigned * flag,
const char ** cigar,
ReadClippingType clipping
)
{
char *field[nSAMFields];
size_t fieldLength[nSAMFields];
if (!parseLine(line, endOfBuffer, field, lineLength, fieldLength)) {
WriteErrorMessage( "Failed to parse SAM line:\n%.*s\n", lineLength, line);
soft_exit(1);
}
//
// We have to copy the contig name (RNAME) into its own buffer because the code in Genome expects
// it to be a null-terminated string, while all we've got is one that's space delimited.
//
const size_t contigNameBufferSize = 512;
char contigName[contigNameBufferSize];
GenomeLocation locationOfContig;
parseContigName(genome, contigName, contigNameBufferSize, &locationOfContig, NULL, field, fieldLength);
GenomeLocation genomeLocation = parseLocation(locationOfContig, field, fieldLength);
if (NULL != out_genomeLocation) {
*out_genomeLocation = genomeLocation;
}
if (fieldLength[SEQ] != fieldLength[QUAL]) {
WriteErrorMessage("SAMReader: QUAL string unequal in length to SEQ string.\n");
soft_exit(1);
}
unsigned _flag;
const size_t flagBufferSize = 20; // More than enough
char flagBuffer[flagBufferSize];
if (fieldLength[FLAG] >= flagBufferSize) {
WriteErrorMessage("SAMReader: flag field is too long.\n");
soft_exit(1);
}
memcpy(flagBuffer,field[FLAG],fieldLength[FLAG]);
flagBuffer[fieldLength[FLAG]] = '\0';
if (1 != sscanf(flagBuffer,"%d",&_flag)) {
WriteErrorMessage("SAMReader: couldn't parse FLAG field.\n");
soft_exit(1);
}
if (NULL != read) {
//
// Clip reads where the quality strings end in '#'
//
unsigned originalFrontClipping, originalBackClipping, originalFrontHardClipping, originalBackHardClipping;
Read::computeClippingFromCigar(field[CIGAR], &originalFrontClipping, &originalBackClipping, &originalFrontHardClipping, &originalBackHardClipping);
unsigned pnext = atoi(field[PNEXT]); // Relies on atoi() returning 0 for non-numeric fields (i.e., *)
read->init(field[QNAME],(unsigned)fieldLength[QNAME],field[SEQ],field[QUAL],(unsigned)fieldLength[SEQ], genomeLocation, atoi(field[MAPQ]), _flag,
originalFrontClipping, originalBackClipping, originalFrontHardClipping, originalBackHardClipping, field[RNEXT], (unsigned)fieldLength[RNEXT], pnext);
//
// If this read is RC in the SAM file, we need to reverse it here, since Reads are always the sense that they were as they came
// out of the base caller.
//
if (_flag & SAM_REVERSE_COMPLEMENT) {
read->becomeRC();
}
read->clip(clipping);
if (field[OPT] != NULL) {
unsigned n = (unsigned) fieldLength[OPT];
while (n > 0 && (field[OPT][n-1] == '\n' || field[OPT][n-1] == '\r')) {
n--;
}
read->setAuxiliaryData(field[OPT], n);
for (char* p = field[OPT]; p != NULL && p < field[OPT] + fieldLength[OPT]; p = SAMReader::skipToBeyondNextFieldSeparator(p, field[OPT] + fieldLength[OPT])) {
if (strncmp(p, "RG:Z:", 5) == 0) {
read->setReadGroup(READ_GROUP_FROM_AUX);
break;
}
}
}
}
if (NULL != alignmentResult) {
if (_flag & SAM_UNMAPPED) {
*alignmentResult = NotFound;
} else {
if ('*' == contigName[0]) {
WriteErrorMessage("SAMReader: mapped read didn't have RNAME filled in.\n");
soft_exit(1);
}
*alignmentResult = SingleHit; // NB: This isn't quite right, we should look at MAPQ.
}
}
if (NULL != direction) {
*direction = (_flag & SAM_REVERSE_COMPLEMENT) ? RC : FORWARD;
}
if (NULL != mapQ) {
*mapQ = atoi(field[MAPQ]);
if (*mapQ > 255) {
WriteErrorMessage("SAMReader: MAPQ field has bogus value\n");
soft_exit(1);
}
}
if (NULL != flag) {
*flag = _flag;
}
if (NULL != cigar) {
*cigar = field[CIGAR];
}
}
void
SAMReader::parseContigName(
const Genome* genome,
char* contigName,
size_t contigNameBufferSize,
GenomeLocation* o_locationOfContig,
int* o_indexOfContig,
char* field[],
size_t fieldLength[],
unsigned rfield)
{
if (fieldLength[rfield] >= contigNameBufferSize) { // >= because we need a byte for the \0
WriteErrorMessage("SAMReader: too long an RNAME. Can't parse.\n");
soft_exit(1);
}
memcpy(contigName,field[rfield],fieldLength[rfield]);
contigName[fieldLength[rfield]] = '\0';
*o_locationOfContig = 0;
if ('*' != contigName[0] && genome != NULL && !genome->getLocationOfContig(contigName, o_locationOfContig, o_indexOfContig)) {
//WriteErrorMessage("Unable to find contig '%s' in genome. SAM file malformed.\n",contigName);
//soft_exit(1);
}
}
GenomeLocation
SAMReader::parseLocation(
GenomeLocation locationOfContig,
char* field[],
size_t fieldLength[],
unsigned rfield,
unsigned posfield)
{
unsigned oneBasedOffsetWithinContig = 0;
if ('*' != field[rfield][0] && '*' != field[posfield][0]) {
//
// We can't call sscanf directly into the mapped file, becuase it reads to the end of the
// string even when it's satisfied all of its fields. Since this can be gigabytes, it's not
// really good for perf. Instead, copy the POS field into a local buffer and null terminate it.
//
const unsigned posBufferSize = 20;
char posBuffer[posBufferSize];
if (fieldLength[posfield] >= posBufferSize) {
WriteErrorMessage("SAMReader: POS field too long.\n");
soft_exit(1);
}
memcpy(posBuffer,field[posfield],fieldLength[posfield]);
posBuffer[fieldLength[posfield]] = '\0';
if (0 == sscanf(posBuffer,"%d",&oneBasedOffsetWithinContig)) {
WriteErrorMessage("SAMReader: Unable to parse position when it was expected.\n");
soft_exit(1);
}
if (0 == oneBasedOffsetWithinContig) {
WriteErrorMessage("SAMReader: Position parsed as 0 when it was expected.\n");
soft_exit(1);
}
return locationOfContig + oneBasedOffsetWithinContig - 1; // -1 is because our offset is 0 based, while SAM is 1 based.
} else {
return InvalidGenomeLocation;
}
}
void
SAMReader::init(
const char *fileName,
_int64 startingOffset,
_int64 amountOfFileToProcess)
{
if (! data->init(fileName)) {
WriteErrorMessage( "Unable to read file %s\n", fileName);
soft_exit(1);
}
if (0 == startingOffset) {
readHeader(fileName);
}
headerSize = context.headerBytes;
reinit(max(startingOffset, (_int64) context.headerBytes),
amountOfFileToProcess == 0 || startingOffset >= (_int64) context.headerBytes ? amountOfFileToProcess
: amountOfFileToProcess - (context.headerBytes - startingOffset));
}
void
SAMReader::reinit(_int64 startingOffset, _int64 amountOfFileToProcess)
{
_ASSERT(-1 != headerSize && startingOffset >= headerSize); // Must call init() before reinit()
//
// There's no way to tell if we start at the very beginning of a read, we need to see the previous newline.
// So, read one byte before our assigned read in case that was the terminating newline of the previous read.
//
if (startingOffset > headerSize) {
startingOffset--;
amountOfFileToProcess++;
}
data->reinit(startingOffset, amountOfFileToProcess);
char* buffer;
_int64 validBytes;
if (!data->getData(&buffer, &validBytes)) {
return;
}
if (startingOffset != headerSize) {
char *firstNewline = strnchr(buffer,'\n',validBytes);
if (NULL == firstNewline) {
return;
}
data->advance((unsigned)(firstNewline - buffer + 1)); // +1 skips over the newline.
}
}
bool
SAMReader::getNextRead(
Read *read,
AlignmentResult *alignmentResult,
GenomeLocation *genomeLocation,
Direction *direction,
unsigned *mapQ,
unsigned *flag,
bool ignoreEndOfRange,
const char **cigar)
{
unsigned local_flag;
if (NULL == flag) {
flag = &local_flag;
}
do {
char* buffer;
_int64 bytes;
if (! data->getData(&buffer, &bytes)) {
data->nextBatch();
if (! data->getData(&buffer, &bytes)) {
return false;
}
}
char *newLine = strnchr(buffer, '\n', bytes);
if (NULL == newLine) {
//
// There is no newline, so the line crosses the end of the buffer.
// This should never happen since underlying reader manages overflow between chunks.
//
WriteErrorMessage("SAM file has too long a line, or doesn't end with a newline! Failing. fileOffset = %lld\n", data->getFileOffset());
soft_exit(1);
}
size_t lineLength;
read->setReadGroup(context.defaultReadGroup);
getReadFromLine(context.genome, buffer,buffer + bytes, read, alignmentResult, genomeLocation, direction, mapQ, &lineLength, flag, cigar, clipping);
read->setBatch(data->getBatch());
data->advance((newLine + 1) - buffer);
} while ((context.ignoreSecondaryAlignments && ((*flag) & SAM_SECONDARY)) ||
(context.ignoreSupplementaryAlignments && ((*flag) & SAM_SUPPLEMENTARY)));
return true;
}
ReadSupplierGenerator *
SAMReader::createReadSupplierGenerator(
const char *fileName,
int numThreads,
const ReaderContext& context)
{
//
// single-ended SAM files always can be read with the range splitter, unless reading from stdin, which needs a queue
//
if (!strcmp(fileName, "-")) {
//
// Stdin must run from a queue, not range splitter.
//
ReadReader* reader;
//
// Because we can only have one stdin reader, we need to use a queue if we're reading from stdin
//
reader = SAMReader::create(DataSupplier::Stdio, "-", ReadSupplierQueue::BufferCount(numThreads), context, 0, 0);
if (reader == NULL) {
return NULL;
}
ReadSupplierQueue *queue = new ReadSupplierQueue(reader);
queue->startReaders();
return queue;
} else {
RangeSplitter *splitter = new RangeSplitter(QueryFileSize(fileName), numThreads, 100);
return new RangeSplittingReadSupplierGenerator(fileName, true, numThreads, context);
}
}
PairedReadReader*
SAMReader::createPairedReader(
const DataSupplier* supplier,
const char *fileName,
int bufferCount,
_int64 startingOffset,
_int64 amountOfFileToProcess,
bool quicklyDropUnpairedReads,
const ReaderContext& context)
{
DataSupplier *data;
if (!strcmp("-", fileName)) {
data = DataSupplier::Stdio;
} else {
data = DataSupplier::Default;
}
SAMReader* reader = SAMReader::create(data, fileName, bufferCount + PairedReadReader::MatchBuffers, context, 0, 0);
if (reader == NULL) {
return NULL;
}
return PairedReadReader::PairMatcher(reader, quicklyDropUnpairedReads);
}
PairedReadSupplierGenerator *
SAMReader::createPairedReadSupplierGenerator(
const char *fileName,
int numThreads,
bool quicklyDropUnpairedReads,
const ReaderContext& context)
{
//
// need to use a queue so that pairs can be matched
//
PairedReadReader* paired = SAMReader::createPairedReader(DataSupplier::Default, fileName,
ReadSupplierQueue::BufferCount(numThreads), 0, 0, quicklyDropUnpairedReads, context);
if (paired == NULL) {
WriteErrorMessage( "Cannot create reader on %s\n", fileName);
soft_exit(1);
}
ReadSupplierQueue* queue = new ReadSupplierQueue(paired);
queue->startReaders();
return queue;
}
const FileFormat* FileFormat::SAM[] = { new SAMFormat(false), new SAMFormat(true) };
void
SAMFormat::getSortInfo(
const Genome* genome,
char* buffer,
_int64 bytes,
GenomeLocation* o_location,
GenomeDistance* o_readBytes,
int* o_refID,
int* o_pos) const
{
char* fields[SAMReader::nSAMFields];
size_t lengths[SAMReader::nSAMFields];
size_t lineLength;
SAMReader::parseLine(buffer, buffer + bytes, fields, &lineLength, lengths);
_ASSERT(lineLength < UINT32_MAX);
if (o_readBytes != NULL) {
*o_readBytes = (unsigned) lineLength;
}
if (lengths[SAMReader::POS] == 0 || fields[SAMReader::POS][0] == '*') {
if (lengths[SAMReader::PNEXT] == 0 || fields[SAMReader::PNEXT][0] == '*') {
if (o_location != NULL) {
*o_location = UINT32_MAX;
}
if (o_refID != NULL) {
*o_refID = -1;
}
if (o_pos != NULL) {
*o_pos = 0;
}
} else {
const size_t contigNameBufferSize = 512;
char contigName[contigNameBufferSize];
GenomeLocation locationOfContig;
SAMReader::parseContigName(genome, contigName, contigNameBufferSize, &locationOfContig, o_refID, fields, lengths, SAMReader::RNEXT);
if (o_location != NULL) {
*o_location = SAMReader::parseLocation(locationOfContig, fields, lengths, SAMReader::RNEXT, SAMReader::PNEXT);
}
}
} else {
const size_t contigNameBufferSize = 512;
char contigName[contigNameBufferSize];
GenomeLocation locationOfContig;
SAMReader::parseContigName(genome, contigName, contigNameBufferSize, &locationOfContig, o_refID, fields, lengths);
if (o_location != NULL) {
*o_location = SAMReader::parseLocation(locationOfContig, fields, lengths);
}
}
}
// which @RG line fields to put in aux data of every read
const char* FileFormat::RGLineToAux = "IDLBPLPUSM";
void
FileFormat::setupReaderContext(
AlignerOptions* options,
ReaderContext* readerContext,
bool bam)
{
if (options->rgLineContents == NULL || *options->rgLineContents == '\0') {
readerContext->defaultReadGroupAux = "";
readerContext->defaultReadGroupAuxLen = 0;
return;
}
char* buffer = new char[strlen(options->rgLineContents) * 3]; // can't expend > 2x
const char* from = options->rgLineContents;
char* to = buffer;
// skip @RG
_ASSERT(strncmp(from, "@RG", 3) == 0);
while (*from && *from != '\t') {
from++;
}
while (*from) {
if (!(from[0] == '\t' && from[1] && from[1] != '\t' && from[2] && from[2] != '\t' && from[3] == ':')) {
WriteErrorMessage("Invalid @RG line: %s\n", options->rgLineContents);
soft_exit(1);
}
bool keep = false;
bool isID = false;
for (const char* a = RGLineToAux; *a; a += 2) {
if (from[1] == a[0] && from[2] == a[1]) {
keep = true;
isID = from[1] == 'I' && from[2] == 'D';
break;
}
}
if (keep) {
if (bam) {
BAMAlignAux* aux = (BAMAlignAux*)to;
aux->tag[0] = isID ? 'R' : from[1];
aux->tag[1] = isID ? 'G' : from[2];
aux->val_type = 'Z';
from += 4; // skip \tXX:
to = (char*)aux->value();
while (*from && *from != '\t') {
*to++ = *from++;
}
*to++ = 0;
} else {
// turn \tXX: into \tXX:Z:, change ID to RG
*to++ = *from++;
if (isID) {
*to++ = 'R';
*to++ = 'G';
from += 2;
} else {
*to++ = *from++;
*to++ = *from++;
}
*to++ = *from++;
*to++ = 'Z';
*to++ = ':';
// copy string attribute
while (*from && *from != '\t') {
*to++ = *from++;
}
}
} else {
from += 4;
while (*from && *from != '\t') {
from++;
}
}
}
readerContext->defaultReadGroupAux = buffer;
readerContext->defaultReadGroupAuxLen = (int) (to - buffer);
}
ReadWriterSupplier*
SAMFormat::getWriterSupplier(
AlignerOptions* options,
const Genome* genome) const
{
DataWriterSupplier* dataSupplier;
if (options->sortOutput) {
size_t len = strlen(options->outputFile.fileName);
// todo: this is going to leak, but there's no easy way to free it, and it's small...
char* tempFileName = (char*) malloc(5 + len);
strcpy(tempFileName, options->outputFile.fileName);
strcpy(tempFileName + len, ".tmp");
dataSupplier = DataWriterSupplier::sorted(this, genome, tempFileName, options->sortMemory * (1ULL << 30),
options->numThreads, options->outputFile.fileName, NULL, options->writeBufferSize);
} else {
dataSupplier = DataWriterSupplier::create(options->outputFile.fileName, options->writeBufferSize);
}
return ReadWriterSupplier::create(this, dataSupplier, genome);
}
bool
SAMFormat::writeHeader(
const ReaderContext& context,
char *header,
size_t headerBufferSize,
size_t *headerActualSize,
bool sorted,
int argc,
const char **argv,
const char *version,
const char *rgLine,
bool omitSQLines) // Hacky option for Charles
const
{
char *commandLine;
size_t commandLineSize = 0;
for (int i = 0; i < argc; i++) {
commandLineSize += strlen(argv[i]) + 1; // +1 is either a space or the terminating null
}
commandLine = new char[commandLineSize];
commandLine[0] = '\0';
for (int i = 0; i < argc; i++) {
strcat(commandLine,argv[i]);
if (i != argc-1) {
strcat(commandLine," ");
}
}
size_t bytesConsumed = snprintf(header, headerBufferSize, "@HD\tVN:1.4\tSO:%s\n%s%s@PG\tID:SNAP\tPN:SNAP\tCL:%s\tVN:%s\n",
sorted ? "coordinate" : "unsorted",
context.header == NULL ? (rgLine == NULL ? "@RG\tID:FASTQ\tSM:sample" : rgLine) : "",
context.header == NULL ? "\n" : "",
commandLine,version);
delete [] commandLine;
commandLine = NULL;
if (bytesConsumed >= headerBufferSize) {
//WriteErrorMessage("SAMWriter: header buffer too small\n");
return false;
}
if (context.header != NULL) {
bool hasRG = false;
for (const char* p = context.header; p < context.header + context.headerLength; ) {
const char* newline = strnchr(p, '\n', (context.header + context.headerLength) - p);
if (newline == NULL) {
newline = context.header + context.headerLength;
}
_ASSERT(newline - p >= 3);
// skip @HD lines, and also @SQ lines if header does not match index
hasRG |= strncmp(p, "@RG", 3) == 0;
if (strncmp(p, "@HD", 3) != 0 &&
(context.headerMatchesIndex || strncmp(p, "@SQ", 3) != 0) &&
strncmp(p, "@PG\tID:SNAP\t", 12) != 0) {
if (bytesConsumed + (newline - p) + 1 >= headerBufferSize) {
//WriteErrorMessage("SAMWriter: header buffer too small\n");
return false;
}
memcpy(header + bytesConsumed, p, (newline - p));
* (header + bytesConsumed + (newline - p)) = '\n';
bytesConsumed += (newline - p) + 1;
}
p = newline + 1;
}
if (! hasRG) {
int n = snprintf(header + bytesConsumed, headerBufferSize - bytesConsumed, "%s\n",
rgLine == NULL ? "@RG\tID:FASTQ\tSM:sample" : rgLine);
if (n > headerBufferSize - bytesConsumed) {
//WriteErrorMessage( "SAMWriter: header buffer too small\n");
return false;
}
bytesConsumed += n;
}
}
#ifndef SKIP_SQ_LINES
if ((context.header == NULL || ! context.headerMatchesIndex) && context.genome != NULL && !omitSQLines) {
// Write an @SQ line for each chromosome / contig in the genome
const Genome::Contig *contigs = context.genome->getContigs();
int numContigs = context.genome->getNumContigs();
GenomeDistance genomeLen = context.genome->getCountOfBases();
size_t originalBytesConsumed = bytesConsumed;
for (int i = 0; i < numContigs; i++) {
GenomeLocation start = contigs[i].beginningLocation;
GenomeLocation end = ((i + 1 < numContigs) ? contigs[i+1].beginningLocation : genomeLen) - context.genome->getChromosomePadding();
bytesConsumed += snprintf(header + bytesConsumed, headerBufferSize - bytesConsumed, "@SQ\tSN:%s\tLN:%u\n", contigs[i].name, end - start);
if (bytesConsumed >= headerBufferSize) {
// todo: increase buffer size (or change to write in batch
bytesConsumed = originalBytesConsumed;
//WriteErrorMessage("SAMWriter: header buffer too small, skipping @SQ lines\n");
return false;
}
}
}
#endif // SKIP_SQ_LINES
*headerActualSize = bytesConsumed;
return true;
}
bool
SAMFormat::createSAMLine(
const Genome * genome,
LandauVishkinWithCigar * lv,
// output data
char* data,
char* quality,
GenomeDistance dataSize,
const char*& contigName,
int& contigIndex,
int& flags,
GenomeDistance& positionInContig,
int& mapQuality,
const char*& matecontigName,
int& mateContigIndex,
GenomeDistance& matePositionInContig,
_int64& templateLength,
unsigned& fullLength,
const char*& clippedData,
unsigned& clippedLength,
unsigned& basesClippedBefore,
unsigned& basesClippedAfter,
// input data
size_t& qnameLen,
Read * read,
AlignmentResult result,
GenomeLocation genomeLocation,
Direction direction,
bool secondaryAlignment,
bool useM,
bool hasMate,
bool firstInPair,
bool alignedAsPair,
Read * mate,
AlignmentResult mateResult,
GenomeLocation mateLocation,
Direction mateDirection,
GenomeDistance *extraBasesClippedBefore)
{
contigName = "*";
positionInContig = 0;
const char *cigar = "*";
templateLength = 0;
if (secondaryAlignment) {
flags |= SAM_SECONDARY;
}
if (0 == qnameLen) {
qnameLen = read->getIdLength();
}
//
// If the aligner said it didn't find anything, treat it as such. Sometimes it will emit the
// best match that it found, even if it's not within the maximum edit distance limit (but will
// then say NotFound). Here, we force that to be SAM_UNMAPPED.
//
if (NotFound == result) {
genomeLocation = InvalidGenomeLocation;
}
if (InvalidGenomeLocation == genomeLocation) {
//
// If it's unmapped, then always emit it in the forward direction. This is necessary because we don't even include
// the SAM_REVERSE_COMPLEMENT flag for unmapped reads, so there's no way to tell that we reversed it.
//
direction = FORWARD;
}
// Write the data and quality strings. If the read is reverse complemented, these need to
// be backwards from the original read. Also, both need to be unclipped.
clippedLength = read->getDataLength();
fullLength = read->getUnclippedLength();
if (fullLength > dataSize) {
return false;
}
if (direction == RC) {
for (unsigned i = 0; i < fullLength; i++) {
data[fullLength - 1 - i] = COMPLEMENT[read->getUnclippedData()[i]];
quality[fullLength - 1 - i] = read->getUnclippedQuality()[i];
}
clippedData = &data[fullLength - clippedLength - read->getFrontClippedLength()];
basesClippedBefore = fullLength - clippedLength - read->getFrontClippedLength();
basesClippedAfter = read->getFrontClippedLength();
} else {
memcpy(data, read->getUnclippedData(), read->getUnclippedLength());
memcpy(quality, read->getUnclippedQuality(), read->getUnclippedLength());
clippedData = read->getData();
basesClippedBefore = read->getFrontClippedLength();
basesClippedAfter = fullLength - clippedLength - basesClippedBefore;
}
int editDistance = -1;
if (genomeLocation != InvalidGenomeLocation) {
if (direction == RC) {
flags |= SAM_REVERSE_COMPLEMENT;
}
const Genome::Contig *contig = genome->getContigForRead(genomeLocation, read->getDataLength(), extraBasesClippedBefore);
_ASSERT(NULL != contig && contig->length > genome->getChromosomePadding());
genomeLocation += *extraBasesClippedBefore;
contigName = contig->name;
contigIndex = (int)(contig - genome->getContigs());
positionInContig = genomeLocation - contig->beginningLocation + 1; // SAM is 1-based
mapQuality = max(0, min(70, mapQuality)); // FIXME: manifest constant.
} else {
flags |= SAM_UNMAPPED;
mapQuality = 0;
*extraBasesClippedBefore = 0;
}
if (hasMate) {
flags |= SAM_MULTI_SEGMENT;
flags |= (firstInPair ? SAM_FIRST_SEGMENT : SAM_LAST_SEGMENT);
if (mateLocation != InvalidGenomeLocation) {
GenomeDistance mateExtraBasesClippedBefore;
const Genome::Contig *mateContig = genome->getContigForRead(mateLocation, mate->getDataLength(), &mateExtraBasesClippedBefore);
mateLocation += mateExtraBasesClippedBefore;
matecontigName = mateContig->name;
mateContigIndex = (int)(mateContig - genome->getContigs());
matePositionInContig = mateLocation - mateContig->beginningLocation + 1;
if (mateDirection == RC) {
flags |= SAM_NEXT_REVERSED;
}
if (genomeLocation == InvalidGenomeLocation) {
//
// The SAM spec says that for paired reads where exactly one end is unmapped that the unmapped
// half should just have RNAME and POS copied from the mate.
//
contigName = matecontigName;
contigIndex = mateContigIndex;
matecontigName = "=";
positionInContig = matePositionInContig;
}
} else {
flags |= SAM_NEXT_UNMAPPED;
//
// The mate's unmapped, so point it at us.
//
matecontigName = "=";
mateContigIndex = contigIndex;
matePositionInContig = positionInContig;
}
if (genomeLocation != InvalidGenomeLocation && mateLocation != InvalidGenomeLocation) {
if (alignedAsPair) {
flags |= SAM_ALL_ALIGNED;
}
// Also compute the length of the whole paired-end string whose ends we saw. This is slightly
// tricky because (a) we may have clipped some bases before/after each end and (b) we need to
// give a signed result based on whether our read is first or second in the pair.
GenomeLocation myStart = genomeLocation - basesClippedBefore;
GenomeLocation myEnd = genomeLocation + clippedLength + basesClippedAfter;
_int64 mateBasesClippedBefore = mate->getFrontClippedLength();
_int64 mateBasesClippedAfter = mate->getUnclippedLength() - mate->getDataLength() - mateBasesClippedBefore;
GenomeLocation mateStart = mateLocation - (mateDirection == RC ? mateBasesClippedAfter : mateBasesClippedBefore);
GenomeLocation mateEnd = mateLocation + mate->getDataLength() + (mateDirection == FORWARD ? mateBasesClippedAfter : mateBasesClippedBefore);
if (contigName == matecontigName) { // pointer (not value) comparison, but that's OK.
if (myStart < mateStart) {
templateLength = mateEnd - myStart;
} else {
templateLength = -(myEnd - mateStart);
}
} // otherwise leave TLEN as zero.
}
if (contigName == matecontigName) {
matecontigName = "="; // SAM Spec says to do this when they're equal (and not *, which won't happen because this is a pointer, not string, compare)
}
}
return true;
}
bool
SAMFormat::writeRead(
const ReaderContext& context,
LandauVishkinWithCigar * lv,
char * buffer,
size_t bufferSpace,
size_t * spaceUsed,
size_t qnameLen,
Read * read,
AlignmentResult result,
int mapQuality,
GenomeLocation genomeLocation,
Direction direction,
bool secondaryAlignment,
int * o_addFrontClipping,
bool hasMate,
bool firstInPair,
Read * mate,
AlignmentResult mateResult,
GenomeLocation mateLocation,
Direction mateDirection,
bool alignedAsPair
) const
{
const int MAX_READ = MAX_READ_LENGTH;
const int cigarBufSize = MAX_READ * 2;
char cigarBuf[cigarBufSize];
const int cigarBufWithClippingSize = MAX_READ * 2 + 32;
char cigarBufWithClipping[cigarBufWithClippingSize];
int flags = 0;
const char *contigName = "*";
int contigIndex = -1;
GenomeDistance positionInContig = 0;
const char *cigar = "*";
const char *matecontigName = "*";
int mateContigIndex = -1;
GenomeDistance matePositionInContig = 0;
_int64 templateLength = 0;
char data[MAX_READ];
char quality[MAX_READ];
const char* clippedData;
unsigned fullLength;
unsigned clippedLength;
unsigned basesClippedBefore;
GenomeDistance extraBasesClippedBefore; // Clipping added if we align before the beginning of a chromosome
unsigned basesClippedAfter;
int editDistance = -1;
*o_addFrontClipping = 0;
if (!createSAMLine(context.genome, lv, data, quality, MAX_READ, contigName, contigIndex,
flags, positionInContig, mapQuality, matecontigName, mateContigIndex, matePositionInContig, templateLength,
fullLength, clippedData, clippedLength, basesClippedBefore, basesClippedAfter,
qnameLen, read, result, genomeLocation, direction, secondaryAlignment, useM,
hasMate, firstInPair, alignedAsPair, mate, mateResult, mateLocation, mateDirection,
&extraBasesClippedBefore))
{
return false;
}
if (genomeLocation != InvalidGenomeLocation) {
cigar = computeCigarString(context.genome, lv, cigarBuf, cigarBufSize, cigarBufWithClipping, cigarBufWithClippingSize,
clippedData, clippedLength, basesClippedBefore, extraBasesClippedBefore, basesClippedAfter,
read->getOriginalFrontHardClipping(), read->getOriginalBackHardClipping(), genomeLocation, direction, useM,
&editDistance, o_addFrontClipping);
if (*o_addFrontClipping != 0) {
return false;
}
}
// Write the SAM entry, which requires the following fields:
//
// 1. QNAME: Query name of the read or the read pair
// 2. FLAG: Bitwise flag (pairing, strand, mate strand, etc.)
// 3. RNAME: Reference sequence name
// 4. POS: 1-Based leftmost position of clipped alignment
// 5. MAPQ: Mapping quality (Phred-scaled)
// 6. CIGAR: Extended CIGAR string (operations: MIDNSHP)
// 7. MRNM: Mate reference name (‘=’ if same as RNAME)
// 8. MPOS: 1-based leftmost mate position
// 9. ISIZE: Inferred insert size
// 10. SEQQuery: Sequence on the same strand as the reference
// 11. QUAL: Query quality (ASCII-33=Phred base quality)
//
// Some FASTQ files have spaces in their ID strings, which is illegal in SAM. Just truncate them at the space.
//
const char *firstSpace = strnchr(read->getId(),' ',qnameLen);
if (NULL != firstSpace) {
qnameLen = (unsigned)(firstSpace - read->getId());
}
const int nmStringSize = 30;// Big enough that it won't buffer overflow regardless of the value of editDistance
char nmString[nmStringSize];
snprintf(nmString, nmStringSize, "\tNM:i:%d",editDistance);
unsigned auxLen;
bool auxSAM;
char* aux = read->getAuxiliaryData(&auxLen, &auxSAM);
static bool warningPrinted = false;
const char* readGroupSeparator = "";
const char* readGroupString = "";
if (aux != NULL && (! auxSAM)) {
if (! warningPrinted) {
WriteErrorMessage( "warning: translating optional fields from BAM->SAM not yet implemented, optional fields will not be included in output\n");
warningPrinted = true;
}
if (read->getReadGroup() == READ_GROUP_FROM_AUX) {
for (BAMAlignAux* bamAux = (BAMAlignAux*) aux; (char*) bamAux < aux + auxLen; bamAux = bamAux->next()) {
if (bamAux->tag[0] == 'R' && bamAux->tag[1] == 'G' && bamAux->val_type == 'Z') {
readGroupSeparator = "\tRG:Z:";
readGroupString = (char*) bamAux->value();
break;
}
}
}
aux = NULL;
auxLen = 0;
}
const char* rglineAux = "";
int rglineAuxLen = 0;
if (read->getReadGroup() != NULL && read->getReadGroup() != READ_GROUP_FROM_AUX) {
if (*readGroupString == 0 || strcmp(readGroupString, context.defaultReadGroup) == 0) {
readGroupSeparator = "";
readGroupString = "";
rglineAux = context.defaultReadGroupAux;
rglineAuxLen = context.defaultReadGroupAuxLen;
} else {
readGroupSeparator = "\tRG:Z:";
readGroupString = read->getReadGroup();
}
}
int charsInString = snprintf(buffer, bufferSpace, "%.*s\t%d\t%s\t%u\t%d\t%s\t%s\t%u\t%lld\t%.*s\t%.*s%s%.*s%s%s\tPG:Z:SNAP%s%.*s\n",
qnameLen, read->getId(),
flags,
contigName,
positionInContig,
mapQuality,
cigar,
matecontigName,
matePositionInContig,
templateLength,
fullLength, data,
fullLength, quality,
aux != NULL ? "\t" : "", auxLen, aux != NULL ? aux : "",
readGroupSeparator, readGroupString,
nmString, rglineAuxLen, rglineAux);
if (charsInString > bufferSpace) {
//
// Out of buffer space.
//
return false;
} else if (charsInString == bufferSpace) {
buffer[bufferSpace-1] = '\n'; // overwrite trailing null with newline
}
if (NULL != spaceUsed) {
*spaceUsed = charsInString;
}
return true;
}
//
// Common cigar string computation between SAM and BAM formats.
//
void
SAMFormat::computeCigar(
CigarFormat cigarFormat,
const Genome * genome,
LandauVishkinWithCigar * lv,
char * cigarBuf,
int cigarBufLen,
const char * data,
GenomeDistance dataLength,
unsigned basesClippedBefore,
GenomeDistance extraBasesClippedBefore,
unsigned basesClippedAfter,
GenomeDistance *o_extraBasesClippedAfter,
GenomeLocation genomeLocation,
bool useM,
int * o_editDistance,
int *o_cigarBufUsed,
int * o_addFrontClipping)
{
if (dataLength > INT32_MAX - MAX_K) {
dataLength = INT32_MAX - MAX_K;
}
int netIndel;
*o_extraBasesClippedAfter = 0;
//
// Apply the extra clipping.
//
genomeLocation += extraBasesClippedBefore;
data += extraBasesClippedBefore;
dataLength -= extraBasesClippedBefore;
const Genome::Contig *contig = genome->getContigAtLocation(genomeLocation);
if (genomeLocation + dataLength > contig->beginningLocation + contig->length - genome->getChromosomePadding()) {
//
// The read hangs off the end of the contig. Soft clip it at the end. This is a tentative amount that assumes no net indels in the
// mapping, we'll refine it later if needed.
//
*o_extraBasesClippedAfter = genomeLocation + dataLength - (contig->beginningLocation + contig->length - genome->getChromosomePadding());
} else {
*o_extraBasesClippedAfter = 0;
}
const char *reference = genome->getSubstring(genomeLocation, dataLength);
if (NULL == reference) {
//
// Fell off the end of the contig.
//
*o_editDistance = 0;
*o_addFrontClipping = 0;
*o_cigarBufUsed = 0;
*cigarBuf = '*';
return;
}
*o_editDistance = lv->computeEditDistanceNormalized(
reference,
(int)(dataLength - *o_extraBasesClippedAfter + MAX_K), // Add space incase of indels. We know there's enough, because the reference is padded.
data,
(int)(dataLength - *o_extraBasesClippedAfter),
MAX_K - 1,
cigarBuf,
cigarBufLen,
useM,
cigarFormat,
o_cigarBufUsed,
o_addFrontClipping,
&netIndel);
if (*o_addFrontClipping != 0) {
//
// On this path, there really isn't a returned cigar string, it's sort of like an exception. We're going up a level and
// trying a different alignment.
//
return;
}
//
// Normally, we'd be done. However, if the amount that we would clip at the end of the read because of hanging off of the end
// of the contig changed, then we need to recompute. In some cases this is an iterative processess as we add or remove bits
// of read.
//
GenomeDistance newExtraBasesClippedAfter = __max(0, genomeLocation + dataLength + netIndel - (contig->beginningLocation + contig->length - genome->getChromosomePadding()));
for (GenomeDistance pass = 0; pass < dataLength; pass++) {
if (newExtraBasesClippedAfter == *o_extraBasesClippedAfter) {
*o_extraBasesClippedAfter = newExtraBasesClippedAfter;
return;
}
*o_extraBasesClippedAfter = newExtraBasesClippedAfter;
*o_editDistance = lv->computeEditDistanceNormalized(
reference,
(int)(dataLength - *o_extraBasesClippedAfter + MAX_K), // Add space incase of indels. We know there's enough, because the reference is padded.
data,
(int)(dataLength - *o_extraBasesClippedAfter),
MAX_K - 1,
cigarBuf,
cigarBufLen,
useM,
cigarFormat,
o_cigarBufUsed,
o_addFrontClipping,
&netIndel);
newExtraBasesClippedAfter = __max(0, genomeLocation + dataLength + netIndel - (contig->beginningLocation + contig->length - genome->getChromosomePadding()));
}
_ASSERT(!"cigar computation didn't converge");
*o_extraBasesClippedAfter = newExtraBasesClippedAfter;
}
// Compute the CIGAR edit sequence string for a read against a given genome location.
// Returns this string if possible or "*" if we fail to compute it (which would likely
// be a bug due to lack of buffer space). The pointer returned may be to cigarBuf so it
// will only be valid until computeCigarString is called again.
const char *
SAMFormat::computeCigarString(
const Genome * genome,
LandauVishkinWithCigar * lv,
char * cigarBuf,
int cigarBufLen,
char * cigarBufWithClipping,
int cigarBufWithClippingLen,
const char * data,
GenomeDistance dataLength,
unsigned basesClippedBefore,
GenomeDistance extraBasesClippedBefore,
unsigned basesClippedAfter,
unsigned frontHardClipping,
unsigned backHardClipping,
GenomeLocation genomeLocation,
Direction direction,
bool useM,
int * o_editDistance,
int * o_addFrontClipping
)
{
GenomeDistance extraBasesClippedAfter;
int cigarBufUsed;
computeCigar(COMPACT_CIGAR_STRING, genome, lv, cigarBuf, cigarBufLen, data, dataLength, basesClippedBefore,
extraBasesClippedBefore, basesClippedAfter, &extraBasesClippedAfter, genomeLocation, useM,
o_editDistance, &cigarBufUsed, o_addFrontClipping);
if (*o_addFrontClipping != 0) {
return NULL;
}
if (*o_editDistance == -2) {
WriteErrorMessage( "WARNING: computeEditDistance returned -2; cigarBuf may be too small\n");
return "*";
} else if (*o_editDistance == -1) {
static bool warningPrinted = false;
if (!warningPrinted) {
WriteErrorMessage( "WARNING: computeEditDistance returned -1; this shouldn't happen\n");
warningPrinted = true;
}
return "*";
} else {
// Add some CIGAR instructions for soft-clipping if we've ignored some bases in the read.
char clipBefore[16] = {'\0'};
char clipAfter[16] = {'\0'};
char hardClipBefore[16] = {'\0'};
char hardClipAfter[16] = {'\0'};
if (frontHardClipping > 0) {
snprintf(hardClipBefore, sizeof(hardClipBefore), "%uH", frontHardClipping);
}
if (basesClippedBefore + extraBasesClippedBefore > 0) {
snprintf(clipBefore, sizeof(clipBefore), "%lluS", basesClippedBefore + extraBasesClippedBefore);
}
if (basesClippedAfter + extraBasesClippedAfter > 0) {
snprintf(clipAfter, sizeof(clipAfter), "%lluS", basesClippedAfter + extraBasesClippedAfter);
}
if (backHardClipping > 0) {
snprintf(hardClipAfter, sizeof(hardClipAfter), "%uH", backHardClipping);
}
snprintf(cigarBufWithClipping, cigarBufWithClippingLen, "%s%s%s%s%s", hardClipBefore, clipBefore, cigarBuf, clipAfter, hardClipAfter);
validateCigarString(genome, cigarBufWithClipping, cigarBufWithClippingLen,
data - basesClippedBefore, dataLength + (basesClippedBefore + basesClippedAfter), genomeLocation + extraBasesClippedBefore, direction, useM);
return cigarBufWithClipping;
}
}
#ifdef _DEBUG
void
SAMFormat::validateCigarString(
const Genome *genome, const char * cigarBuf, int cigarBufLen, const char *data, GenomeDistance dataLength, GenomeLocation genomeLocation, Direction direction, bool useM)
{
const char *nextChunkOfCigar = cigarBuf;
GenomeDistance offsetInData = 0;
const char *reference = genome->getSubstring(genomeLocation, dataLength);
if (NULL == reference) {
WriteErrorMessage("validateCigarString: couldn't look up genome data for location %lld\n", genomeLocation);
soft_exit(1);
}
GenomeDistance offsetInReference = 0;
bool sawNonH = false; // This is to make sure that the clipping types (H & S) occur only at the beginning or end of the cigar string.
bool sawTailS = false; // Did we see a S
bool sawLeadingS = false; // Have we seen the soft clip at the front of the cigar string?
bool sawTrailingH = false;
char previousOp = '\0'; // Make sure that we don't have two consecutive ops of the same type that should be merged
bool sawXorM = false;
bool lastItemWasIndel = false;
//
// First check to see that it's null terminated
//
bool nullTerminated = false;
for (size_t offset = 0; offset < cigarBufLen; offset++) {
if ('\0' == cigarBuf[offset]) {
nullTerminated = true;
break;
}
}
if (!nullTerminated) {
WriteErrorMessage("validateCigarString: non-null-terminated or overflow cigar string: '%.*s'\n", cigarBufLen, cigarBuf);
soft_exit(1);
}
const Genome::Contig *contig = genome->getContigAtLocation(genomeLocation);
if (NULL == contig) {
WriteErrorMessage("validateCigarString: read alignment location isn't in a chromosome, genomeLocation %lld\n", GenomeLocationAsInt64(genomeLocation));
soft_exit(1);
}
if (genomeLocation >= contig->beginningLocation + contig->length - genome->getChromosomePadding()) {
WriteErrorMessage("validateCigarString: alignment location is in genome padding: %lld, contig name %s, base %lld, len %lld, padding size %d\n",
GenomeLocationAsInt64(genomeLocation), contig->name, GenomeLocationAsInt64(contig->beginningLocation), contig->length, genome->getChromosomePadding());
soft_exit(1);
}
while ('\0' != *nextChunkOfCigar) {
unsigned len;
char op;
int fieldsScanned = sscanf(nextChunkOfCigar, "%d%c", &len, &op);
if (2 != fieldsScanned) {
WriteErrorMessage("validateCigarString: didn't scan two fields here '%s' in overall cigar string '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (0 == len) {
WriteErrorMessage("validateCigarString: got zero length field here '%s' in overall cigar string '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (op != 'H' && sawTailS) {
WriteErrorMessage("validateCigarString: saw incorrect op type after what should have been the terminal soft or hard clipping here '%s', in overall cigar string '%s'\n",
nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (sawTrailingH) {
WriteErrorMessage("validateCigarString: saw op after what should have been the terminal hard clip here '%s' in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (op == previousOp) {
WriteErrorMessage("validateCigarString: saw consecutive ops of the same type '%c' here '%s' in overall cigar '%s'\n", op, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
switch (op) {
case 'M':
{
if (!useM) {
WriteErrorMessage("validateCigarString: generated an M when we were supposed to use X and = here '%s' in overall cigar string '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
offsetInData += len;
sawNonH = true;
sawXorM = true;
lastItemWasIndel = false;
break;
}
case 'X':
case '=':
{
if (useM) {
WriteErrorMessage("validateCigarString: generated an %c when were supposed to use M here '%s' in overall cigar string '%s'\n", op, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (len + offsetInData > dataLength) {
WriteErrorMessage("validateCigarString: cigar string overflowed read length, here '%s', overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
for (unsigned offset = 0; offset < len; offset++) {
if ((data[offset + offsetInData] == reference[offset + offsetInReference]) == ('X' == op)) {
WriteErrorMessage("validateCigarString: saw a (non-)matching base in an %c range, offset %d, offsetInData %lld, offsetInReference %lld, data '%.*s', reference '%.*s', here '%s', overall cigar '%s'\n",
op, offset, offsetInData, offsetInReference, dataLength, data, dataLength, reference, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
}
offsetInData += len;
offsetInReference += len;
sawNonH = true;
sawXorM = true;
lastItemWasIndel = false;
break;
}
case 'I':
{
//
// Insertion uses up bases in the read but not in the reference.
//
if (len + offsetInData > dataLength) {
WriteErrorMessage("validateCigarString: insertion pushes cigar string overlength, here '%s' in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (!sawXorM) {
WriteErrorMessage("validateCigarString: cigar string started with I (after clipping) here '%s' in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (previousOp == 'D') {
WriteErrorMessage("validateCigarString: cigar string had D immediately followed by I here '%'s in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
offsetInData += len;
sawNonH = true;
lastItemWasIndel = true;
break;
}
case 'D':
{
if (!sawXorM) {
WriteErrorMessage("validateCigarString: cigar string started with D (after clipping) here '%s' in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
if (previousOp == 'I') {
WriteErrorMessage("validateCigarString: cigar string had I immediately followed by D here '%'s in overall cigar '%s'\n", nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
//
// D uses up bases in the reference but not the read.
//
offsetInReference += len;
sawNonH = true;
lastItemWasIndel = true;
break;
}
case 'N':
case 'P':
{
WriteErrorMessage("validateCigarString: saw valid op type '%c' that SNAP shouldn't generate, here '%s' in overall cigar string '%s'\n", op, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
case 'H':
{
//
// Hard clip bases do not occur in the read string at all. All we can validate is that this is the first or last thing in the cigar string.
//
if (nextChunkOfCigar == cigarBuf) {
//
// First thing, this is OK.
//
break;
}
sawTrailingH = true;
break;
}
case 'S':
{
if (sawNonH) {
sawTailS = true;
}
sawNonH = true;
offsetInData += len;
break;
}
default: {
WriteErrorMessage("validateCigarString: got unrecognized cigar op '%c', here '%s' in overall string '%s'\n", op, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
}
previousOp = op;
//
// Now scan over the current op.
//
while ('0' <= *nextChunkOfCigar && '9' >= *nextChunkOfCigar) {
nextChunkOfCigar++;
}
if (*nextChunkOfCigar != op) {
WriteErrorMessage("validateCigarString: bug in validation code; expected op '%c', got '%c' at '%s' in '%s'\n", op, *nextChunkOfCigar, nextChunkOfCigar, cigarBuf);
soft_exit(1);
}
nextChunkOfCigar++;
}
if (offsetInData != dataLength) {
WriteErrorMessage("validateCigarString: Didn't consume entire read data, got %lld of %lld, cigar '%s'\n", offsetInData, dataLength, cigarBuf);
soft_exit(1);
}
if (lastItemWasIndel) {
WriteErrorMessage("validateCigarString: cigar string ended with indel '%s'\n", cigarBuf);
soft_exit(1);
}
//
// Make sure none of the non-soft-clipped part of the read is mapped onto padding.
//
if (genomeLocation + offsetInReference > contig->beginningLocation + contig->length - genome->getChromosomePadding()) {
WriteErrorMessage("validateCigarString: alignment runs into contig padding: %lld, contig name %s, base %lld, len %lld, padding size %d, offsetInReference %lld\n",
GenomeLocationAsInt64(genomeLocation), contig->name, GenomeLocationAsInt64(contig->beginningLocation), contig->length, genome->getChromosomePadding(), offsetInReference);
soft_exit(1);
}
}
#endif // _DEBUG
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