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/*++
Module Name:
FASTQ.cpp
Abstract:
Fast FASTQ genome "query" reader.
Authors:
Bill Bolosky, August, 2011
Environment:
User mode service.
This class is NOT thread safe. It's the caller's responsibility to ensure that
at most one thread uses an instance at any time.
Revision History:
Adapted from Matei Zaharia's Scala implementation.
--*/
#include "stdafx.h"
#include "FASTQ.h"
#include "Compat.h"
#include "BigAlloc.h"
#include "Read.h"
#include "Util.h"
#include "exit.h"
#include "Error.h"
using std::min;
using util::strnchr;
FASTQReader::FASTQReader(
DataReader* i_data,
const ReaderContext& i_context)
:
ReadReader(i_context),
data(i_data)
{
}
FASTQReader::~FASTQReader()
{
delete data;
data = NULL;
}
FASTQReader*
FASTQReader::create(
DataSupplier* supplier,
const char *fileName,
int bufferCount,
_int64 startingOffset,
_int64 amountOfFileToProcess,
const ReaderContext& context)
{
DataReader* data = supplier->getDataReader(bufferCount, maxReadSizeInBytes, 0.0, 0);
FASTQReader* fastq = new FASTQReader(data, context);
if (! fastq->init(fileName)) {
WriteErrorMessage("Unable to initialize FASTQReader for file %s\n", fileName);
soft_exit(1);
}
fastq->reinit(startingOffset, amountOfFileToProcess);
return fastq;
}
void
FASTQReader::readHeader(
const char* fileName,
ReaderContext& context)
{
// no header in FQ files
context.header = NULL;
context.headerLength = context.headerBytes = 0;
}
bool
FASTQReader::init(
const char* i_fileName)
{
fileName = i_fileName;
return data->init(fileName);
}
void
FASTQReader::reinit(
_int64 startingOffset,
_int64 amountOfFileToProcess)
{
data->reinit(startingOffset, amountOfFileToProcess);
char* buffer;
_int64 bytes;
if (! data->getData(&buffer, &bytes)) {
return;
}
// If we're not at the start of the file, we might have the tail end of a read that someone
// who got the previous range will process; advance past that. This is fairly tricky because
// there can be '@' signs in the quality string (and maybe even in read names?).
if (startingOffset != 0) {
if (!skipPartialRecord(data)) {
//
// There wasn't a whole record in our range. Skip over what we had.
//
data->advance(bytes);
return;
}
}
}
bool
FASTQReader::skipPartialRecord(DataReader *data)
{
//
// Just assume that a single FASTQ read is smaller than our buffer, so we won't exceed the buffer here.
// Look for the pattern '{0|\n}@*\n{A|T|C|G|N}*\n+' That is, either the beginning of the buffer or a
// newline, followed by an '@' and some text, another newline followed by a list of bases and a newline,
// and then a plus.
//
char* buffer;
_int64 validBytes;
data->getData(&buffer, &validBytes);
char *firstLineCandidate = buffer;
if (*firstLineCandidate != '@') {
firstLineCandidate = strnchr(buffer, '\n', validBytes) + 1;
}
for (;;) {
if (firstLineCandidate - buffer >= validBytes) {
// This happens for very small files.
return false;
}
char *secondLineCandidate = strnchr(firstLineCandidate, '\n', validBytes - (firstLineCandidate - buffer)) + 1;
if (NULL == (secondLineCandidate-1)) {
WriteErrorMessage("Unable to find a read in FASTQ buffer (2) at %d\n", data->getFileOffset());
return false;
}
if (*firstLineCandidate != '@') {
firstLineCandidate = secondLineCandidate;
continue;
}
//
// Scan through the second line making sure it's all bases (or 'N'). We don't have to
// check for end-of-buffer because we know there's a null there.
//
char *thirdLineCandidate = secondLineCandidate;
while (*thirdLineCandidate == 'A' || *thirdLineCandidate == 'C' || *thirdLineCandidate == 'T' || *thirdLineCandidate == 'G' ||
*thirdLineCandidate == 'N' || *thirdLineCandidate == 'a' || *thirdLineCandidate == 'c' || *thirdLineCandidate == 't' ||
*thirdLineCandidate == 'g' || *thirdLineCandidate == 'n') {
thirdLineCandidate++;
}
if (*thirdLineCandidate == '\r') {
//
// CRLF text; skip the CR.
//
thirdLineCandidate++;
}
if (*thirdLineCandidate != '\n') {
//
// We found something that's not a base and not a newline. It wasn't a read data (second) line. Move up a line
// and try again.
//
firstLineCandidate = secondLineCandidate;
continue;
}
thirdLineCandidate++;
if (*thirdLineCandidate != '+') {
firstLineCandidate = secondLineCandidate;
continue;
}
break;
}
data->advance(firstLineCandidate - buffer);
return true;
}
//
// Try to parse a read starting at a given position pos, updating readToUpdate with it.
// Returns 0 if the parse failed or the first position past the read if it succeeds. In
// addition, if exitOnFailure is set, print a warning and exit if there is a parse error.
// (The one time we don't set this is when trying to find the first read in a chunk.)
//
bool
FASTQReader::getNextRead(Read *readToUpdate)
{
//
// Find the next newline.
//
char* buffer;
_int64 validBytes;
if (! data->getData(&buffer, &validBytes)) {
data->nextBatch();
if (! data->getData(&buffer, &validBytes)) {
return false;
}
}
_int64 bytesConsumed = getReadFromBuffer(buffer, validBytes, readToUpdate, fileName, data, context);
if (bytesConsumed == 0) {
return false;
}
data->advance(bytesConsumed);
return true;
}
//static char LAST[100000]; static int LASTLEN = 0;
_int64
FASTQReader::getReadFromBuffer(char *buffer, _int64 validBytes, Read *readToUpdate, const char *fileName, DataReader *data, const ReaderContext &context)
{
//
// Get the next four lines.
//
char* lines[nLinesPerFastqQuery];
unsigned lineLengths[nLinesPerFastqQuery];
char* scan = buffer;
for (unsigned i = 0; i < nLinesPerFastqQuery; i++) {
char *newLine = strnchr(scan, '\n', validBytes - (scan - buffer));
if (NULL == newLine) {
if (validBytes - (scan - buffer) == 1 && *scan == 0x1a && data->isEOF()) {
// sometimes DOS files will have extra ^Z at end
return false;
}
//
// There was no next newline
//
if (data->isEOF()) {
WriteErrorMessage("FASTQ file doesn't end with a newline! Failing. fileOffset = %lld, validBytes = %d\n",
data->getFileOffset(),validBytes);
soft_exit(1);
}
WriteErrorMessage("FASTQ record larger than buffer size at %s:%lld\n", fileName, data->getFileOffset());
soft_exit(1);
}
const size_t lineLen = newLine - scan;
if (0 == lineLen) {
WriteErrorMessage("Syntax error in FASTQ file: blank line.\n");
soft_exit(1);
}
if (! isValidStartingCharacterForNextLine[(i + 3) % 4][*scan]) {
WriteErrorMessage("FASTQ file %s has invalid starting character at offset %lld, line type %d, char %c\n", data->getFilename(), data->getFileOffset(), i, *scan);
WriteErrorMessage("Line in question: '%.*s'\n", lineLen, scan);
//WriteErrorMessage("Preceding record: '%.*s'\n", LASTLEN, LAST);
soft_exit(1);
}
lines[i] = scan;
lineLengths[i] = (unsigned) lineLen - (scan[lineLen-1] == '\r' ? 1 : 0);
scan = newLine + (newLine[1] == '\r' ? 2 : 1);
}
const char *id = lines[0] + 1; // The '@' on the first line is not part of the ID
const char* space = strnchr(id, ' ', lineLengths[0] - 1);
readToUpdate->init(id, space != NULL ? (unsigned) (space - id) : (unsigned) lineLengths[0] - 1, lines[1], lines[3], lineLengths[1]);
readToUpdate->clip(context.clipping);
readToUpdate->setBatch(data->getBatch());
readToUpdate->setReadGroup(context.defaultReadGroup);
//memcpy(LAST, buffer, scan - buffer); LASTLEN = scan - buffer;
return scan - buffer;
}
//
// static data & initialization
//
bool FASTQReader::isValidStartingCharacterForNextLine[FASTQReader::nLinesPerFastqQuery][256];
FASTQReader::_init FASTQReader::_initializer;
FASTQReader::_init::_init()
{
//
// Initialize the isValidStartingCharacterForNextLine array.
//
memset(isValidStartingCharacterForNextLine, 0, sizeof(isValidStartingCharacterForNextLine));
//
// The first line is the descriptor line and must start with an '@'
//
isValidStartingCharacterForNextLine[3]['@'] = true;
//
// The second line is the read itself and must start with a base or an
// 'N' in either case. A . is just a different way to encode an N.
//
for (const char*p = "ACTGNURYKMSWBDHVNX."; *p; p++) {
isValidStartingCharacterForNextLine[0][*p] = true;
isValidStartingCharacterForNextLine[0][tolower(*p)] = true;
}
//
//
// The third line is additional sequence idenfitier info and must
// start with a '+'.
//
isValidStartingCharacterForNextLine[1]['+'] = true;
//
// Line 4 is the quality line. It starts with a printable ascii character.
// It would be nice to rule out the bases, N, + and @ because it might confsue the parser,
// but some quality lines do start with those...
//
for (char i = '!'; i <= '~'; i++) {
isValidStartingCharacterForNextLine[2][i] = true;
}
}
//
// PairedInterleavedFASTQReader
//
PairedInterleavedFASTQReader::PairedInterleavedFASTQReader(
DataReader* i_data,
const ReaderContext& i_context) :
data(i_data), context(i_context)
{
}
PairedInterleavedFASTQReader*
PairedInterleavedFASTQReader::create(DataSupplier* supplier, const char *fileName, int bufferCount, _int64 startingOffset, _int64 amountOfFileToProcess,
const ReaderContext& context)
{
DataReader* data = supplier->getDataReader(bufferCount, 2 * maxReadSizeInBytes, 0.0, 0); // 2* because we read in pairs
PairedInterleavedFASTQReader* fastq = new PairedInterleavedFASTQReader(data, context);
if (! fastq->init(fileName)) {
WriteErrorMessage("Unable to initialize PairedInterleavedFASTQReader for file %s\n", fileName);
soft_exit(1);
}
fastq->reinit(startingOffset, amountOfFileToProcess);
return fastq;
}
bool
PairedInterleavedFASTQReader::init(const char* i_fileName)
{
fileName = i_fileName;
return data->init(fileName);
}
bool
PairedInterleavedFASTQReader::getNextReadPair(Read *read0, Read *read1)
{
//
// Find the next newline.
//
char* buffer;
_int64 validBytes, startBytes;
if (! data->getData(&buffer, &validBytes, &startBytes)) {
//fprintf(stderr, "!getData offset %lld\n", data->getFileOffset());
data->nextBatch();
//fprintf(stderr, "FQ batch\n");
if (! data->getData(&buffer, &validBytes, &startBytes)) {
return false;
}
//fprintf(stderr, "nextBatch getData offset %lld\n", data->getFileOffset());
//fprintf(stderr, "FQ data %.10s, start ...%.10s, valid ...%.10s, %lld bytes\n", buffer, buffer+validBytes-10, buffer+startBytes-10, validBytes);
}
_int64 bytesConsumed = FASTQReader::getReadFromBuffer(buffer, validBytes, read0, fileName, data, context);
if (bytesConsumed == validBytes) {
WriteErrorMessage("Input file seems to have an odd number of reads. Ignoring the last one.");
return false;
}
bytesConsumed += FASTQReader::getReadFromBuffer(buffer + bytesConsumed, validBytes - bytesConsumed, read1, fileName, data, context);
//
// Validate the Read IDs.
//
if (read0->getIdLength() < 2 || memcmp(read0->getId() + read0->getIdLength() - 2, "/1", 2)) {
WriteErrorMessage("PairedInterleavedFASTQReader: first read of batch doesn't have ID ending with /1: '%.*s'\n", read0->getIdLength(), read0->getId());
soft_exit(1);
}
if (read1->getIdLength() < 2 || memcmp(read1->getId() + read1->getIdLength() - 2, "/2", 2)) {
WriteErrorMessage("PairedInterleavedFASTQReader: second read of batch doesn't have ID ending with /2: '%.*s'\n", read1->getIdLength(), read1->getId());
soft_exit(1);
}
data->advance(bytesConsumed);
return true;
}
void
PairedInterleavedFASTQReader::reinit(_int64 startingOffset, _int64 amountOfFileToProcess)
{
data->reinit(startingOffset, amountOfFileToProcess);
char* buffer;
_int64 bytes;
if (! data->getData(&buffer, &bytes)) {
return;
}
// If we're not at the start of the file, we might have the tail end of a read that someone
// who got the previous range will process; advance past that. This is fairly tricky because
// there can be '@' signs in the quality string (and maybe even in read names?).
if (startingOffset != 0) {
if (!FASTQReader::skipPartialRecord(data)) {
return;
}
}
//
// Grab the first read from the buffer, and see if it's /1 or /2.
//
if (!data->getData(&buffer, &bytes)) {
return;
}
Read read;
_int64 bytesForFirstRead = FASTQReader::getReadFromBuffer(buffer, bytes, &read, fileName, data, context);
if (read.getIdLength() < 2 || read.getId()[read.getIdLength() - 2] != '/' || (read.getId()[read.getIdLength() - 1] != '1' && read.getId()[read.getIdLength() -1] != '2') ) {
WriteErrorMessage("PairedInterleavedFASTQReader: read ID doesn't appear to end with /1 or /2, you can't use this as a paired FASTQ file: '%.*s'\n", read.getIdLength(), read.getId());
soft_exit(1);
}
if (read.getId()[read.getIdLength()-1] == '2') {
//
// This is the second half of a pair. Skip it.
//
data->advance(bytesForFirstRead);
//
// Now make sure that the next read is /1.
//
if (!data->getData(&buffer, &bytes)) {
//
// This was the last read in the file.
//
return;
}
FASTQReader::getReadFromBuffer(buffer, bytes, &read, fileName, data, context);
if (read.getIdLength() < 2 || read.getId()[read.getIdLength()-2] != '/' || read.getId()[read.getIdLength()-1] != '1') {
WriteErrorMessage("PairedInterleavedFASTQReader: first read of pair doesn't appear to have an ID that ends in /1: '%.*s'\n", read.getIdLength(), read.getId());
soft_exit(1);
}
}
}
//
// FASTQWriter
//
FASTQWriter *
FASTQWriter::Factory(const char *filename)
{
FILE *file = fopen(filename,"wb");
if (NULL == file) {
return NULL;
}
return new FASTQWriter(file);
}
bool
FASTQWriter::writeRead(Read *read)
{
size_t len = read->getIdLength() + 2 * (read->getDataLength() + 2) /* @ and + */ + 10 /* crlf + padding + null */;
if (bufferSize - bufferOffset <= len) {
flushBuffer();
}
size_t bytesUsed = snprintf(buffer + bufferOffset, bufferSize - bufferOffset, "@%.*s\n%.*s\n+\n%.*s\n", read->getIdLength(), read->getId(), read->getDataLength(), read->getData(), read->getDataLength(), read->getQuality());
bufferOffset += bytesUsed;
return true;
}
PairedFASTQReader::~PairedFASTQReader()
{
for (int i = 0; i < 2; i++) {
delete readers[i];
readers[i] = NULL;
}
}
PairedFASTQReader *
PairedFASTQReader::create(
DataSupplier* supplier,
const char *fileName0,
const char *fileName1,
int bufferCount,
_int64 startingOffset,
_int64 amountOfFileToProcess,
const ReaderContext& context)
{
PairedFASTQReader *reader = new PairedFASTQReader;
reader->readers[0] = FASTQReader::create(supplier, fileName0, bufferCount, startingOffset, amountOfFileToProcess, context);
reader->readers[1] = FASTQReader::create(supplier, fileName1, bufferCount, startingOffset, amountOfFileToProcess, context);
for (int i = 0; i < 2; i++) {
if (NULL == reader->readers[i]) {
delete reader;
reader = NULL;
return NULL;
}
}
return reader;
}
bool
PairedFASTQReader::getNextReadPair(Read *read0, Read *read1)
{
bool worked = readers[0]->getNextRead(read0);
if (readers[1]->getNextRead(read1) != worked) {
WriteErrorMessage("PairedFASTQReader: reads of both ends responded differently. The FASTQ files may not match properly.\n");
soft_exit(1);
}
return worked;
}
PairedReadSupplierGenerator *
PairedFASTQReader::createPairedReadSupplierGenerator(
const char *fileName0,
const char *fileName1,
int numThreads,
const ReaderContext& context,
bool gzip)
{
const char *fileNames[2] = {fileName0, fileName1};
//
// Decide whether to use the range splitter or a queue based on whether the files are the same size.
//
if (!strcmp("-", fileNames[0]) || !strcmp("-", fileNames[1]) || QueryFileSize(fileNames[0]) != QueryFileSize(fileNames[1]) || gzip) {
//WriteStatusMessage("FASTQ using supplier queue\n");
DataSupplier* dataSupplier[2];
size_t fileSize[2];
for (int i = 0; i < 2; i++) {
if (!strcmp(fileNames[i], "-")) {
fileSize[i] = 0;
if (gzip) {
dataSupplier[i] = DataSupplier::GzipStdio;
} else {
dataSupplier[i] = DataSupplier::Stdio;
}
} else {
fileSize[i] = QueryFileSize(fileNames[i]);
if (gzip) {
dataSupplier[i] = DataSupplier::GzipDefault;
} else {
dataSupplier[i] = DataSupplier::Default;
}
}
}
int bufferCount = ReadSupplierQueue::BufferCount(numThreads);
ReadReader *reader1 = FASTQReader::create(dataSupplier[0], fileName0, bufferCount, 0, fileSize[0],context);
ReadReader *reader2 = FASTQReader::create(dataSupplier[1], fileName1, bufferCount, 0, fileSize[1],context);
if (NULL == reader1 || NULL == reader2) {
delete reader1;
delete reader2;
return NULL;
}
ReadSupplierQueue *queue = new ReadSupplierQueue(reader1,reader2);
queue->startReaders();
return queue;
} else {
//WriteStatusMessage("FASTQ using range splitter\n");
return new RangeSplittingPairedReadSupplierGenerator(fileName0, fileName1, FASTQFile, numThreads, false, context);
}
}
ReadSupplierGenerator *
FASTQReader::createReadSupplierGenerator(
const char *fileName,
int numThreads,
const ReaderContext& context,
bool gzip)
{
bool isStdin = !strcmp(fileName,"-");
if (! gzip && !isStdin) {
//
// Single ended uncompressed FASTQ files can be handled by a range splitter.
//
return new RangeSplittingReadSupplierGenerator(fileName, false, numThreads, context);
} else {
ReadReader* fastq;
//
// Because we can only have one stdin reader, we need to use a queue if we're reading from stdin
//
if (isStdin) {
if (gzip) {
fastq = FASTQReader::create(DataSupplier::GzipStdio, fileName, ReadSupplierQueue::BufferCount(numThreads), 0, 0, context);
} else {
fastq = FASTQReader::create(DataSupplier::Stdio, fileName, ReadSupplierQueue::BufferCount(numThreads), 0, 0, context);
}
} else {
fastq = FASTQReader::create(DataSupplier::GzipDefault, fileName, ReadSupplierQueue::BufferCount(numThreads), 0, QueryFileSize(fileName), context);
}
if (fastq == NULL) {
delete fastq;
return NULL;
}
ReadSupplierQueue *queue = new ReadSupplierQueue(fastq);
queue->startReaders();
return queue;
}
}
PairedReadSupplierGenerator *
PairedInterleavedFASTQReader::createPairedReadSupplierGenerator(
const char *fileName,
int numThreads,
const ReaderContext& context,
bool gzip)
{
bool isStdin = !strcmp(fileName,"-");
if (gzip || isStdin) {
//WriteStatusMessage("PairedInterleavedFASTQ using supplier queue\n");
DataSupplier *dataSupplier;
if (isStdin) {
if (gzip) {
dataSupplier = DataSupplier::GzipStdio;
} else {
dataSupplier = DataSupplier::Stdio;
}
} else {
dataSupplier = DataSupplier::GzipDefault;
}
PairedReadReader *reader = PairedInterleavedFASTQReader::create(dataSupplier, fileName,
ReadSupplierQueue::BufferCount(numThreads), 0,(stdin ? 0 : QueryFileSize(fileName)),context);
if (NULL == reader ) {
delete reader;
return NULL;
}
ReadSupplierQueue *queue = new ReadSupplierQueue(reader);
queue->startReaders();
return queue;
} else {
//WriteStatusMessage("PairedInterleavedFASTQ using range splitter\n");
return new RangeSplittingPairedReadSupplierGenerator(fileName, NULL, InterleavedFASTQFile, numThreads, false, context);
}
}
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