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#define __FAST_MATH__
#include <algorithm>
#include <cassert>
#include <cstdio>
#include <map>
#include <numeric>
#include <set>
#include <string>
#include <alignment/datastructures/alignment/ByteAlignment.h>
#include <alignment/datastructures/alignment/AlignmentMap.hpp>
#include <alignment/datastructures/alignment/CmpFile.hpp>
#include <alignment/files/BaseSequenceIO.hpp>
#include <alignment/utils/FileOfFileNames.hpp>
#include <hdf/HDFBasReader.hpp>
#include <hdf/HDFCCSReader.hpp>
#include <hdf/HDFCmpFile.hpp>
#include <hdf/HDFPlsReader.hpp>
#include <pbdata/CommandLineParser.hpp>
#include <pbdata/alignment/CmpAlignment.hpp>
#include <pbdata/loadpulses/MetricField.hpp>
#include <pbdata/loadpulses/MovieAlnIndexLookupTable.hpp>
#include <pbdata/reads/BaseFile.hpp>
#include <pbdata/reads/PulseFile.hpp>
#include <pbdata/reads/ReadType.hpp>
#include <pbdata/utils/TimeUtils.hpp>
typedef std::map<std::string, int> MovieNameToArrayIndex;
typedef std::map<std::string, bool> MetricOptionsMap;
typedef std::map<std::string, std::vector<std::string> > RequirementMap;
char VERSION[] = "v1.1.0";
char PERFORCE_VERSION_STRING[] = "$Change: 126407 $";
// define default values for metrics
const float NaN = 0.0 / 0.0;
const UChar missingQualityValue = 255;
const unsigned char maxQualityValue = 100;
const HalfWord missingFrameRateValue = USHRT_MAX;
const unsigned int missingPulseIndex = UINT_MAX;
void CapQualityValue(QualityValueVector<QualityValue> &vect, DNALength length,
unsigned char maxQualityValue = 100)
{
unsigned int i;
if (vect.data == NULL) {
return;
}
for (i = 0; i < length; i++) {
vect.data[i] = std::min(vect.data[i], maxQualityValue);
}
}
void CapQualityValues(SMRTSequence &seq, unsigned char maxQualityValue = 100)
{
CapQualityValue(seq.qual, seq.length, maxQualityValue);
CapQualityValue(seq.deletionQV, seq.length, maxQualityValue);
CapQualityValue(seq.preBaseDeletionQV, seq.length, maxQualityValue);
CapQualityValue(seq.insertionQV, seq.length, maxQualityValue);
CapQualityValue(seq.substitutionQV, seq.length, maxQualityValue);
CapQualityValue(seq.mergeQV, seq.length, maxQualityValue);
}
int CheckCmpFileFormat(CmpFile &cmpFile)
{
if (cmpFile.readType != ReadType::Standard) {
std::cout << "ERROR! Reading pulse information into a cmp.h5 file generated from circular "
<< std::endl
<< "consensus called sequences is not supported." << std::endl;
std::exit(EXIT_FAILURE);
}
return 1;
}
void BuildRequirementMap(RequirementMap &fieldRequirements)
{
fieldRequirements["StartTimeOffset"].push_back("StartFrame");
fieldRequirements["StartTimeOffset"].push_back("NumEvent");
fieldRequirements["StartFrame"].push_back("PreBaseFrames");
fieldRequirements["StartFrame"].push_back("WidthInFrames");
fieldRequirements["PulseWidth"].push_back("WidthInFrames");
fieldRequirements["pkmid"].push_back("MidSignal");
fieldRequirements["pkmid"].push_back("NumEvent");
fieldRequirements["IPD"].push_back("StartFrame");
fieldRequirements["IPD"].push_back("NumEvent");
fieldRequirements["IPD"].push_back("PreBaseFrames");
fieldRequirements["IPD"].push_back("WidthInFrames");
fieldRequirements["Light"].push_back("MeanSignal");
fieldRequirements["Light"].push_back("NumEvent");
fieldRequirements["Light"].push_back("WidthInFrames");
// Build requirementMap for sneaky metrics
fieldRequirements["StartFrameBase"].push_back("PreBaseFrames");
fieldRequirements["StartFrameBase"].push_back("WidthInFrames");
fieldRequirements["StartFramePulse"].push_back("PreBaseFrames");
fieldRequirements["StartFramePulse"].push_back("WidthInFrames");
}
void ExclusivelyAdd(const char *value, std::vector<std::string> &vect)
{
if (find(vect.begin(), vect.end(), value) == vect.end()) {
vect.push_back(value);
}
}
bool AnyFieldRequiresFrameRate(std::vector<std::string> &fields)
{
for (size_t i = 0; i < fields.size(); i++) {
if (fields[i] == "PulseWidth" or fields[i] == "IPD" or fields[i] == "Light" or
fields[i] == "StartTimeOffset" or fields[i] == "StartFrame" or
fields[i] == "PulseWidth" or fields[i] == "PreBaseFrames" or
fields[i] == "WidthInFrames") {
return true;
}
}
return false;
}
template <typename T>
void Free(T *&buf)
{
if (buf != NULL) {
delete[] buf;
}
buf = NULL;
}
// Return all eighteen metrics that can be loaded.
// StartTimeOffset QualityValue InsertionQV MergeQV
// DeletionQV DeletionTag PulseIndex SubstitutionTag
// SubstitutionQV ClassifierQV StartFrame PulseWidth
// PreBaseFrames WidthInFrames pkmid IPD
// Light WhenStarted
std::vector<std::string> GetAllSupportedMetrics(bool isSneakyMetricsIncluded = true)
{
// The order of metrics matters. With -bymetric option, all fields
// which are required for computing a metric are cached before WriteMetric()
// and cleared afterwards. If two neighboring metrics share a subset of
// required fields, then the cached fields can be re-used. Arrange metrics
// in an order that maximizes reuse of cached fields.
std::vector<std::string> supportedMetrics;
supportedMetrics.push_back("WhenStarted");
supportedMetrics.push_back("QualityValue");
supportedMetrics.push_back("InsertionQV");
supportedMetrics.push_back("MergeQV");
supportedMetrics.push_back("DeletionQV");
supportedMetrics.push_back("DeletionTag");
supportedMetrics.push_back("SubstitutionTag");
supportedMetrics.push_back("SubstitutionQV");
supportedMetrics.push_back("PreBaseFrames");
// Sneaky metrics for internal use Only
if (isSneakyMetricsIncluded) {
supportedMetrics.push_back("StartFrameBase");
}
supportedMetrics.push_back("IPD");
supportedMetrics.push_back("StartFrame");
if (isSneakyMetricsIncluded) {
supportedMetrics.push_back("StartFramePulse");
}
// Disable metric StartTimeOffset for now.
// StartTimeOffset is placed at the same level as AlnArray, However, the
// size of StartTimeOffset is far less than AlnArray, while cmp.h5 spec
// requires all datasets at that level to have the same size.
// supportedMetrics.push_back("StartTimeOffset");
supportedMetrics.push_back("PulseWidth");
supportedMetrics.push_back("WidthInFrames");
supportedMetrics.push_back("Light");
supportedMetrics.push_back("pkmid");
supportedMetrics.push_back("ClassifierQV");
supportedMetrics.push_back("PulseIndex");
return supportedMetrics;
}
// Return metrics to load by default.
std::vector<std::string> GetDefaultMetrics()
{
std::vector<std::string> defaultMetrics;
defaultMetrics.push_back("QualityValue");
defaultMetrics.push_back("ClassifierQV");
defaultMetrics.push_back("StartFrame");
defaultMetrics.push_back("PulseWidth");
defaultMetrics.push_back("WidthInFrames");
defaultMetrics.push_back("pkmid");
defaultMetrics.push_back("IPD");
return defaultMetrics;
}
// Return metrics that can be computed from PulseCalls.
std::vector<std::string> GetPulseMetrics()
{
std::vector<std::string> pulseMetrics;
pulseMetrics.push_back("StartFrame");
pulseMetrics.push_back("StartTimeOffset");
pulseMetrics.push_back("ClassifierQV");
pulseMetrics.push_back("PulseWidth");
pulseMetrics.push_back("WidthInFrames");
pulseMetrics.push_back("IPD");
pulseMetrics.push_back("pkmid");
pulseMetrics.push_back("Light");
pulseMetrics.push_back("StartFramePulse");
return pulseMetrics;
}
// Return true if this metric can be computed from PulseCalls.
bool IsPulseMetric(const std::string &metric)
{
std::vector<std::string> pulseMetrics = GetPulseMetrics();
for (size_t i = 0; i < pulseMetrics.size(); i++) {
if (pulseMetrics[i] == metric) return true;
}
return false;
}
// Return all metrics that are
// (1) supported,
// (2) requested to load, and
// (3) computable with all required fields available
// in either bas.h5 or pls.h5.
std::vector<std::string> GetMetricsToLoad(std::map<std::string, bool> &metricOptions)
{
std::vector<std::string> metricsToLoad;
// Get all supported metrics.
std::vector<std::string> supportedMetrics = GetAllSupportedMetrics();
std::map<std::string, bool>::iterator metricIt;
for (size_t i = 0; i < supportedMetrics.size(); i++) {
std::string metric = supportedMetrics[i];
metricIt = metricOptions.find(metric);
if (metricIt != metricOptions.end() and metricIt->second) {
// Get metrics that are required and computable
metricsToLoad.push_back(metricIt->first);
}
}
return metricsToLoad;
}
void StoreDatasetFieldsFromPulseFields(MetricOptionsMap &fieldSet,
RequirementMap &fieldRequirements,
std::vector<std::string> &datasetFields)
{
size_t d;
MetricOptionsMap::iterator optionsIt;
for (optionsIt = fieldSet.begin(); optionsIt != fieldSet.end(); ++optionsIt) {
if (optionsIt->second == true) {
if (fieldRequirements.find(optionsIt->first) == fieldRequirements.end()) {
ExclusivelyAdd(optionsIt->first.c_str(), datasetFields);
} else {
for (d = 0; d < fieldRequirements[optionsIt->first].size(); d++) {
ExclusivelyAdd(fieldRequirements[optionsIt->first][d].c_str(), datasetFields);
}
}
}
}
}
void ParseMetricsList(std::string metricListString, MetricOptionsMap &metricOptions)
{
std::vector<std::string> metrics;
Splice(metricListString, ",", metrics);
for (size_t m = 0; m < metrics.size(); m++) {
if (metricOptions.find(metrics[m]) != metricOptions.end()) {
metricOptions[metrics[m]] = true;
} else {
std::cout << "ERROR! Metric " << metrics[m] << " is not supported." << std::endl;
std::exit(EXIT_FAILURE);
}
}
}
// Set default metric options to true
void SetDefaultMetricOptions(std::map<std::string, bool> &metricOptions)
{
std::vector<std::string> defaultMetrics = GetDefaultMetrics();
for (size_t i = 0; i < defaultMetrics.size(); i++) {
metricOptions[defaultMetrics[i]] = true;
}
}
// Initialize all supported metric options and set all to false
void CreateMetricOptions(std::map<std::string, bool> &metricOptions)
{
std::vector<std::string> supportedMetrics = GetAllSupportedMetrics();
for (size_t i = 0; i < supportedMetrics.size(); i++) {
metricOptions[supportedMetrics[i]] = false;
}
}
// Check whether all fields are available or not.
bool AreAllFieldsAvailable(std::vector<Field> &requiredFields, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, const bool &useBaseFile,
const bool &usePulseFile)
{
bool allAvailable = true;
for (size_t i = 0; i < requiredFields.size(); i++) {
Field field = requiredFields[i];
if (field.type == BasField) {
if (!useBaseFile or !hdfBasReader.FieldIsIncluded(field.name) or
!hdfBasReader.includedFields[field.name]) {
allAvailable = false;
break;
}
} else if (field.type == PlsField) {
if (!usePulseFile or !hdfPlsReader.FieldIsIncluded(field.name) or
!hdfPlsReader.includedFields[field.name]) {
allAvailable = false;
break;
}
}
}
return allAvailable;
}
//
// Check whether a metric is computable or not.
// fieldsToBeUsed = all fields that will be used for computing a metric.
// If a metric can be computed from both bas and pls files (e.g.
// StartFrame, IPD, PulseWidth, WidthInFrame), only compute it from pls.
//
bool CanThisMetricBeComputed(const std::string &metricName, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, const bool &useBaseFile,
const bool &usePulseFile, std::vector<Field> &fieldsToBeUsed)
{
fieldsToBeUsed.clear();
FieldsRequirement fieldsRequirement = FieldsRequirement(metricName);
bool metricMayBeComputedFromPls = true;
if (fieldsRequirement.fieldsUsePlsFile.size() != 0 && usePulseFile) {
metricMayBeComputedFromPls =
AreAllFieldsAvailable(fieldsRequirement.fieldsUsePlsFile, hdfBasReader, hdfPlsReader,
useBaseFile, usePulseFile);
} else {
metricMayBeComputedFromPls = false;
}
bool metricMayBeComputedFromBas = true;
if (fieldsRequirement.fieldsUseBasFile.size() != 0 && useBaseFile) {
metricMayBeComputedFromBas =
AreAllFieldsAvailable(fieldsRequirement.fieldsUseBasFile, hdfBasReader, hdfPlsReader,
useBaseFile, usePulseFile);
} else {
metricMayBeComputedFromBas = false;
}
bool metricMayBeComputed = true;
if (!metricMayBeComputedFromBas and !metricMayBeComputedFromPls) {
metricMayBeComputed = false;
}
// Compute from pls if possible
if (metricMayBeComputedFromPls) {
fieldsToBeUsed = fieldsRequirement.fieldsUsePlsFile;
} else if (metricMayBeComputedFromBas) {
fieldsToBeUsed = fieldsRequirement.fieldsUseBasFile;
}
if (metricName == "StartTimeOffset") {
metricMayBeComputed = false;
// Disable StartTimeOffset for now.
}
if (metricName == "WhenStarted") {
// WhenStarted requires no fields from neither bas nor pls.
metricMayBeComputed = true;
}
return metricMayBeComputed;
}
//
// Check whether metrics are computable or not. If a metric is not
// computable, disable it with a warning or exit with an error.
//
void CanMetricsBeComputed(MetricOptionsMap &metricOptions, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, const bool &useBaseFile,
const bool &usePulseFile, const bool &failOnMissingData,
const std::string &movieName)
{
std::map<std::string, bool>::iterator metricIt;
for (metricIt = metricOptions.begin(); metricIt != metricOptions.end(); ++metricIt) {
std::string metricName = metricIt->first;
if (metricName == "") {
metricIt->second = false;
}
if (metricIt->second == false) {
continue;
}
std::vector<Field> fieldsToBeUsed;
bool metricMayBeComputed = CanThisMetricBeComputed(
metricName, hdfBasReader, hdfPlsReader, useBaseFile, usePulseFile, fieldsToBeUsed);
if (metricMayBeComputed == false) {
if (failOnMissingData) {
std::cout << "ERROR";
} else {
std::cout << "WARNING";
}
std::cout << ": There is insufficient data to compute metric: " << metricName
<< " in the file " << movieName << " ";
std::cout << " It will be ignored." << std::endl;
if (failOnMissingData) {
std::exit(EXIT_FAILURE);
}
metricOptions[metricName] = false;
}
}
}
// Return size of a single field in KB.
UInt ComputeRequiredMemoryForThisField(Field &thisField, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, const bool &
#ifndef NDEBUG
useBaseFile
#endif
,
const bool &
#ifndef NDEBUG
usePulseFile
#endif
)
{
if (thisField.type == BasField) {
assert(useBaseFile);
return hdfBasReader.GetFieldSize(thisField.name);
}
if (thisField.type == PlsField) {
assert(usePulseFile);
return hdfPlsReader.GetFieldSize(thisField.name);
}
throw std::runtime_error{"Unknown Field type"};
}
//
// Return estimated memory peak (in KB) for buffering all data using -bymetric.
//
UInt ComputeRequiredMemory(std::vector<std::string> &metricsToLoad, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, const bool &useBaseFile,
const bool &usePulseFile, HDFCmpFile<CmpAlignment> &cmpReader,
UInt &totalAlnLength)
{
UInt maxMemory = 0;
for (size_t i = 0; i < metricsToLoad.size(); i++) {
UInt memoryForThisMetric = 0;
std::vector<Field> fieldsToBeUsed;
bool canBeComputed = CanThisMetricBeComputed(metricsToLoad[i], hdfBasReader, hdfPlsReader,
useBaseFile, usePulseFile, fieldsToBeUsed);
(void)(canBeComputed);
for (size_t j = 0; j < fieldsToBeUsed.size(); j++) {
UInt memoryForThisField = ComputeRequiredMemoryForThisField(
fieldsToBeUsed[j], hdfBasReader, hdfPlsReader, useBaseFile, usePulseFile);
memoryForThisMetric += memoryForThisField;
}
maxMemory = std::max(maxMemory, memoryForThisMetric);
}
//
// AlnIndex will be buffered. Some other datastructures also need
// to be buffered for quick look up. Approximately double the size.
//
UInt totalAlnIndexMem = 2 * cmpReader.alnInfoGroup.GetAlnIndexSize();
//
// AlnArray and metrics to load needs to be buffered in KB.
//
UInt totalAlnArrayMem = totalAlnLength / 1024 * (sizeof(unsigned int) + sizeof(unsigned char));
//
// It's diffcult to estimate how much memory will be used by hdf5.
// Assume memory consumed by hdf5 scales with AlnIndex and AlnArray datasets.
//
UInt hdf5Mem = totalAlnIndexMem / 2 + totalAlnLength / 1024 * sizeof(unsigned int);
maxMemory += totalAlnIndexMem + totalAlnArrayMem + hdf5Mem;
//std::cout << "The estimated peak memory for buffering fields is "
// << maxMemory << " KB." << std::endl;
//std::cout << "The estimated memory for buffering AlnIndex related data is "
// << totalAlnIndexMem << " KB."<< std::endl;
//std::cout << "The estimated memory for buffering AlnArray related data is "
// << totalAlnArrayMem << " KB." << std::endl;
//std::cout << "The estimated memory for hdf5 is "
// << hdf5Mem << " KB." << std::endl;
//std::cout << "The estimated total memory is "
// << maxMemory << " KB." << std::endl;
return maxMemory;
}
//
// Get aligned sequence for this alignment from cmpFile
//
std::string GetAlignedSequenceFromCmpFile(const HDFCmpFile<CmpAlignment> &cmpReader,
MovieAlnIndexLookupTable &lookupTable)
{
std::string alignedSequence;
std::vector<unsigned char> byteAlignment;
int alignedSequenceLength = lookupTable.offsetEnd - lookupTable.offsetBegin;
if (alignedSequenceLength >= 0) {
alignedSequence.resize(alignedSequenceLength);
byteAlignment.resize(alignedSequenceLength);
}
//
// Read the alignment string. All alignments
//
cmpReader.refAlignGroups[lookupTable.refGroupIndex]
->readGroups[lookupTable.readGroupIndex]
->alignmentArray.Read(lookupTable.offsetBegin, lookupTable.offsetEnd, &byteAlignment[0]);
//
// Convert to something we can compare easily.
//
ByteAlignmentToQueryString(&byteAlignment[0], byteAlignment.size(), &alignedSequence[0]);
return alignedSequence;
}
//
// Store info necessary for loading pulses to lookupTable.
//
void BuildLookupTable(const int &movieAlignmentIndex, CmpFile &cmpFile, BaseFile &baseFile,
const bool &usePulseFile, PulseFile &pulseFile,
HDFCmpFile<CmpAlignment> &cmpReader, const std::vector<int> &movieAlnIndex,
const std::vector<std::pair<int, int> > &toFrom,
const std::set<uint32_t> &moviePartHoleNumbers,
MovieAlnIndexLookupTable &lookupTable)
{
//
// Query the cmp file for a way to look up a read based on
// coordinate information. For Astro reads, the coords are
// based on x and y. For Springfield, it is read index. The
// base files should be able to look up reads by x,y or by
// index.
//
if (cmpFile.platformId == Astro) {
std::cout << "ASTRO pulse loading is deprecated." << std::endl;
std::exit(EXIT_FAILURE);
}
int alignmentIndex = movieAlnIndex[toFrom[movieAlignmentIndex].second];
//
// Alignments are grouped by ref group id then movie id.
//
int refGroupId = cmpFile.alnInfo.alignments[alignmentIndex].GetRefGroupId();
int movieId = cmpFile.alnInfo.alignments[alignmentIndex].GetMovieId();
(void)(movieId);
UInt holeNumber = cmpFile.alnInfo.alignments[alignmentIndex].GetHoleNumber();
int alnGroupId = cmpFile.alnInfo.alignments[alignmentIndex].GetAlnGroupId();
if (cmpReader.refGroupIdToArrayIndex.find(refGroupId) ==
cmpReader.refGroupIdToArrayIndex.end()) {
std::cout << "ERROR! An alignment " << alignmentIndex
<< " is specified with reference group " << std::endl
<< refGroupId << " that is not found as an alignment group." << std::endl;
std::exit(EXIT_FAILURE);
}
int refGroupIndex = cmpReader.refGroupIdToArrayIndex[refGroupId];
//
// Now find the group containing the alignment.
//
if (cmpReader.alnGroupIdToReadGroupName.find(alnGroupId) ==
cmpReader.alnGroupIdToReadGroupName.end()) {
std::cout << "ERROR! An alignment " << alignmentIndex
<< " is specified with alignment group " << std::endl
<< alnGroupId << " that is not found." << std::endl;
std::exit(EXIT_FAILURE);
}
std::string readGroupName = cmpReader.alnGroupIdToReadGroupName[alnGroupId];
if (cmpReader.refAlignGroups[refGroupIndex]->experimentNameToIndex.find(readGroupName) ==
cmpReader.refAlignGroups[refGroupIndex]->experimentNameToIndex.end()) {
std::cout << "ERROR! An alignment " << alignmentIndex
<< " is specified with read group name " << std::endl
<< readGroupName << " that is not found." << std::endl;
std::exit(EXIT_FAILURE);
}
int readGroupIndex =
cmpReader.refAlignGroups[refGroupIndex]->experimentNameToIndex[readGroupName];
UInt offsetBegin = cmpFile.alnInfo.alignments[alignmentIndex].GetOffsetBegin();
UInt offsetEnd = cmpFile.alnInfo.alignments[alignmentIndex].GetOffsetEnd();
//
// First pull out the bases corresponding to this read.
//
int queryStart = cmpFile.alnInfo.alignments[alignmentIndex].GetQueryStart();
int queryEnd = cmpFile.alnInfo.alignments[alignmentIndex].GetQueryEnd();
bool skip = false;
int readIndex, readStart, readLength, plsReadIndex;
readIndex = readStart = readLength = plsReadIndex = -1;
//
// Since the movie may be split into multiple parts, look to see
// if this hole number is one of the ones covered by this
// set. If it is not, just continue. It will be loaded on
// another pass through a different movie part.
//
if (moviePartHoleNumbers.find(holeNumber) == moviePartHoleNumbers.end()) {
skip = true;
} else {
if (!baseFile.LookupReadIndexByHoleNumber(holeNumber, readIndex)) {
std::cout << "ERROR! Alignment has hole number " << holeNumber
<< " that is not in the movie. " << std::endl;
std::exit(EXIT_FAILURE);
}
readStart = baseFile.readStartPositions[readIndex];
readLength =
baseFile.readStartPositions[readIndex + 1] - baseFile.readStartPositions[readIndex];
if (usePulseFile) {
if (!pulseFile.LookupReadIndexByHoleNumber(holeNumber, plsReadIndex)) {
std::cout << "ERROR! Alignment has hole number " << holeNumber
<< " that is not in the movie. " << std::endl;
std::exit(EXIT_FAILURE);
}
assert(pulseFile.holeNumbers[plsReadIndex] == baseFile.holeNumbers[readIndex]);
}
}
// Save info to lookupTable
lookupTable.SetValue(skip, // Skip processing this or not
movieAlignmentIndex, alignmentIndex, refGroupIndex, readGroupIndex,
holeNumber, // cmp.h5 /AlnInfo/AlnIndex column 7
offsetBegin, // cmp.h5 /AlnInfo/AlnIndex column 18
offsetEnd, // cmp.h5 /AlnInfo/AlnIndex column 19
queryStart, // cmp.h5 /AlnInfo/AlnIndex column 11
queryEnd, // cmp.h5 /AlnInfo/AlnIndex column 12
readIndex, // hole Index in BaseCalls/ZMW/HoleNumber
readStart, // readStart in BaseCalls/* (e.g. *=Basecall)
readLength, // readLength in BaseCalls/*
plsReadIndex); // readIndex in PulseCalls/ZMW/HoleNumber
}
//
// Map bases of a read to pulse indices.
//
void MapBaseToPulseIndex(BaseFile &baseFile, PulseFile &pulseFile, MovieAlnIndexLookupTable &table,
std::vector<int> &baseToPulseIndexMap)
{
baseToPulseIndexMap.resize(table.readLength);
int pulseStart = pulseFile.pulseStartPositions[table.plsReadIndex];
//
// Copy the subset of pulses that correspond to the ones called as bases.
//
int i;
for (i = 0; i < table.readLength; i++) {
baseToPulseIndexMap[i] = pulseStart + baseFile.pulseIndex[table.readStart + i];
}
}
//
// Get source read from the bas/pls file.
//
void GetSourceRead(CmpFile &cmpFile, BaseFile &baseFile, PulseFile &pulseFile,
HDFBasReader &hdfBasReader, HDFPlsReader &hdfPlsReader,
HDFCCSReader<SMRTSequence> &hdfCcsReader, const bool &useBaseFile,
const bool &usePulseFile, const bool &useCcsOnly,
//const bool & byRead,
MovieAlnIndexLookupTable &table, const std::string &alignedSequence,
SMRTSequence &sourceRead, unsigned int &numPasses)
{
(void)(baseFile);
(void)(pulseFile);
(void)(alignedSequence);
assert(!table.skip);
//
// These are not allocated in the regular allocate function
// since they are only used in loadPulses. (maybe I should
// subclass SMRTSequence here).
//
//if (byRead) {
// Read in the data from the bas file if it exsts.
if (useBaseFile) {
hdfBasReader.GetReadAt(table.readIndex, sourceRead);
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
numPasses = hdfCcsReader.GetNumPasses(table.readIndex);
}
}
// Read in the data from the pls file if it exists.
if (usePulseFile) {
hdfPlsReader.GetReadAt(table.plsReadIndex, sourceRead.pulseIndex, sourceRead);
}
// }
// else {
// This is deprecated
// //
// // The entire base/pulse file was read in, so copy data from that into a read
// // For the data used in the read, it is possible to simply
// // reference the data, but for the pls file it is necessary
// // to copy since there is a packing of data.
// //
// if (useBaseFile) {
// baseFile.CopyReadAt(table.readIndex, sourceRead);
// if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
// numPasses = hdfCcsReader.GetNumPasses(table.readIndex);
// }
// }
// if (usePulseFile) {
// std::vector<int> baseToPulseIndexMap;
// MapBaseToPulseIndex(baseFile, pulseFile, table, baseToPulseIndexMap);
// pulseFile.CopyReadAt(table.readIndex, &baseToPulseIndexMap[0], sourceRead);
// }
//}
CapQualityValues(sourceRead);
}
//
// Build lookup tables for all alignments whose indices in
// AlnArray are saved in movieAlnIndex.
// Also check whether the bas file and the cmp file match.
//
void BuildLookupTablesAndMakeSane(CmpFile &cmpFile, BaseFile &baseFile, PulseFile &pulseFile,
HDFCmpFile<CmpAlignment> &cmpReader, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader,
HDFCCSReader<SMRTSequence> &hdfCcsReader, const bool &useBaseFile,
const bool &usePulseFile, const bool &useCcsOnly,
const std::vector<int> &movieAlnIndex,
const std::vector<std::pair<int, int> > &toFrom,
const std::set<uint32_t> &moviePartHoleNumbers,
std::vector<MovieAlnIndexLookupTable> &lookupTables)
{
(void)(hdfPlsReader);
(void)(hdfCcsReader);
(void)(useCcsOnly);
(void)(useBaseFile);
lookupTables.resize(movieAlnIndex.size());
size_t movieAlignmentIndex = 0;
for (movieAlignmentIndex = 0; movieAlignmentIndex < movieAlnIndex.size();
movieAlignmentIndex++) {
BuildLookupTable(movieAlignmentIndex, cmpFile, baseFile, usePulseFile, pulseFile, cmpReader,
movieAlnIndex, toFrom, moviePartHoleNumbers,
lookupTables[movieAlignmentIndex]);
}
//
// Load entire Basecall from pls/bas to memory, and
// check whether aligned sequences in cmp.h5 matches
// sequences in pls/bas or not
//
hdfBasReader.ReadField(baseFile, "Basecall");
//
// For each alignment, do sanity check and
// cache aligned sequence in MovieAlnIndexLookupTable
//
for (movieAlignmentIndex = 0; movieAlignmentIndex < movieAlnIndex.size();
movieAlignmentIndex++) {
MovieAlnIndexLookupTable &table = lookupTables[movieAlignmentIndex];
if (table.skip) continue;
//
// Get aligned sequence for this alignment from cmpFile
//
std::string alignedSequence = GetAlignedSequenceFromCmpFile(cmpReader, table);
// Save the aligned sequence in the table
table.alignedSequence = alignedSequence;
RemoveGaps(alignedSequence, alignedSequence);
//
// Get sequence for this alignment from baseFile
//
Nucleotide *seq = new Nucleotide[table.readLength];
baseFile.CopyArray(baseFile.baseCalls, table.readStart, table.readLength, seq);
std::string readSequence;
readSequence.resize(table.queryEnd - table.queryStart);
copy((char *)(seq + table.queryStart), (char *)(seq + table.queryEnd),
readSequence.begin());
delete[] seq;
//
// Do a sanity check to make sure the pulses and the alignment
// make sense. The main check is to see if the query sequence
// in the alignment is the same as the query sequence in the
// read.
//
if (alignedSequence.size() != readSequence.size() or alignedSequence != readSequence) {
std::cout << "ERROR, the query sequence does not match the aligned query sequence."
<< std::endl
<< "HoleNumber: "
<< cmpFile.alnInfo.alignments[table.alignmentIndex].GetHoleNumber()
<< ", MovieName: " << baseFile.GetMovieName()
<< ", ReadIndex: " << table.readIndex << ", qStart: " << table.queryStart
<< ", qEnd: " << table.queryEnd << std::endl
<< "Aligned sequence: " << std::endl
<< alignedSequence << std::endl
<< "Original sequence: " << std::endl
<< readSequence << std::endl;
std::exit(EXIT_FAILURE);
}
}
hdfBasReader.ClearField(baseFile, "Basecall");
}
// Given a vector of lookupTables in which items with the same
// refGroupIndex and readGroupIndex are grouped, find index boundaries
// of each group and save these boundaries to groupedLookupTablesIndexPairs
// The index boundary of each group consists of:
// 1, index (0 based, inclusive) of the very first item of a group
// 2, index (0 based, exclusive) of the very last item of a group
//
// Assume that lookupTables satisfy the following criteria.
// 1, items are already grouped by refGroupIndex and readGroupIndex
// 2, items which have the same alnGroupIndex, should have
// the same refGroupIndex and readGroupIndex
// Note that:
// 1, alnGroupIndex represents index of AlnGroupID, (i.e. dataset
// /AlnInfo/AlnIndex column 1);
// refGroupIndex represents index of RefGroupID, (i.e. dataset
// /AlnInfo/AlnIndex column 3);
// readGroupIndex represents index of an experiment group within
// a refGroup (e.g. if a refGroup /ref0001 contains two experiment
// groups /ref0001/movie1 and /ref0001/movie2, then readGroupIndex
// for these two groups are 0 and 1.).
// 2, within each grouped item, offsetBegin may not begin from 0,
// and offsets may not be continugous.
//
void GroupLookupTables(std::vector<MovieAlnIndexLookupTable> &lookupTables,
std::vector<std::pair<UInt, UInt> > &groupedLookupTablesIndexPairs)
{
std::vector<std::pair<UInt, UInt> > refGroupIndexReadGroupIndexPairs;
UInt movieAlignmentIndex = 0;
size_t preRefGroupIndex = 0;
size_t preReadGroupIndex = 0;
UInt pairFirst = 0;
bool isVeryFirstGroup = true;
for (movieAlignmentIndex = 0; movieAlignmentIndex < lookupTables.size();
movieAlignmentIndex++) {
MovieAlnIndexLookupTable &lookupTable = lookupTables[movieAlignmentIndex];
if (isVeryFirstGroup or (lookupTable.refGroupIndex != preRefGroupIndex or
lookupTable.readGroupIndex != preReadGroupIndex)) {
// Find a new group
if (isVeryFirstGroup) {
// This is the very first group
isVeryFirstGroup = false;
} else if (lookupTable.refGroupIndex == preRefGroupIndex &&
lookupTable.readGroupIndex != preReadGroupIndex) {
// Assumption (1) has been violated
std::cout << "ERROR! lookupTables should have been sorted by reference"
<< "group index and read group index." << std::endl;
std::exit(EXIT_FAILURE);
} else {
// Find the first lookupTable of a new group, save indices of [first and last)
// lookupTables of the last group.
groupedLookupTablesIndexPairs.push_back(
std::pair<UInt, UInt>(pairFirst, movieAlignmentIndex));
// Save refGroupIndex and readGroupIndex of the last group
std::pair<UInt, UInt> refGroupIndexReadGroupIndexPair(preRefGroupIndex,
preReadGroupIndex);
refGroupIndexReadGroupIndexPairs.push_back(refGroupIndexReadGroupIndexPair);
}
// Store index of the first lookupTable of the new group in lookupTables
pairFirst = movieAlignmentIndex;
// Store refGroupIndex and readGroupIndex of the new group
preRefGroupIndex = lookupTable.refGroupIndex;
preReadGroupIndex = lookupTable.readGroupIndex;
}
}
if (not isVeryFirstGroup) {
// Save indices of [first and last) lookupTables of the very last group
groupedLookupTablesIndexPairs.push_back(
std::pair<UInt, UInt>(pairFirst, movieAlignmentIndex));
// Save refGroupIndex and readGroupIndex of the very last group
std::pair<UInt, UInt> refGroupIndexReadGroupIndexPair(preRefGroupIndex, preReadGroupIndex);
refGroupIndexReadGroupIndexPairs.push_back(refGroupIndexReadGroupIndexPair);
} // Do nothing, if no lookupTable exists
// Double check all assumptions are met
for (size_t i = 0; i < refGroupIndexReadGroupIndexPairs.size(); i++) {
for (size_t j = i + 1; j < refGroupIndexReadGroupIndexPairs.size(); j++) {
// Assure that assumption (1) is met. If this assertion fails,
// then alignments in the input cmp.h5 are not grouped by
// reference. Check /AlnInfo/AlnIndex dataset column 3.
assert(refGroupIndexReadGroupIndexPairs[i] != refGroupIndexReadGroupIndexPairs[j]);
}
}
assert(groupedLookupTablesIndexPairs.size() == refGroupIndexReadGroupIndexPairs.size());
for (size_t i = 0; i < groupedLookupTablesIndexPairs.size(); i++) {
UInt firstIndex = groupedLookupTablesIndexPairs[i].first;
UInt lastIndex = groupedLookupTablesIndexPairs[i].second;
UInt refGroupIndex = refGroupIndexReadGroupIndexPairs[i].first;
UInt readGroupIndex = refGroupIndexReadGroupIndexPairs[i].second;
for (UInt index = firstIndex; index < lastIndex; index++) {
assert(lookupTables[index].refGroupIndex == refGroupIndex);
assert(lookupTables[index].readGroupIndex == readGroupIndex);
}
}
}
//
// Read all required fields for computing the specified metric into memory,
// unless the fields have been cached.
//
void CacheRequiredFieldsForMetric(BaseFile &baseFile, PulseFile &pulseFile,
HDFBasReader &hdfBasReader, HDFPlsReader &hdfPlsReader,
HDFCCSReader<SMRTSequence> &hdfCcsReader, const bool &useBaseFile,
const bool &usePulseFile, const bool &useCcsOnly,
std::vector<Field> &cachedFields, const std::string &curMetric)
{
(void)(hdfCcsReader);
(void)(useCcsOnly);
std::vector<Field> fieldsToBeUsed;
bool canBeComputed = CanThisMetricBeComputed(curMetric, hdfBasReader, hdfPlsReader, useBaseFile,
usePulseFile, fieldsToBeUsed);
assert(canBeComputed);
// Cache all required fields
for (size_t i = 0; i < fieldsToBeUsed.size(); i++) {
bool isFieldCached = false;
for (size_t j = 0; j < cachedFields.size(); j++) {
if (fieldsToBeUsed[i] == cachedFields[j]) {
isFieldCached = true;
break;
}
}
if (isFieldCached) {
continue;
}
std::string &curField = fieldsToBeUsed[i].name;
FieldType &fieldType = fieldsToBeUsed[i].type;
if (fieldType == BasField and useBaseFile and hdfBasReader.FieldIsIncluded(curField) and
hdfBasReader.includedFields[curField]) {
hdfBasReader.ReadField(baseFile, curField);
cachedFields.push_back(fieldsToBeUsed[i]);
} else if (fieldType == PlsField and usePulseFile and
hdfPlsReader.FieldIsIncluded(curField) and
hdfPlsReader.includedFields[curField]) {
hdfPlsReader.ReadField(pulseFile, curField);
cachedFields.push_back(fieldsToBeUsed[i]);
}
}
}
//
// Clear cached fields unless they are also required for computing
// the next metric.
//
void ClearCachedFields(BaseFile &baseFile, PulseFile &pulseFile, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, HDFCCSReader<SMRTSequence> &hdfCcsReader,
const bool &useBaseFile, const bool &usePulseFile, const bool &useCcsOnly,
std::vector<Field> &cachedFields, const std::string &curMetric,
const std::string &nextMetric)
{
(void)(hdfCcsReader);
(void)(useCcsOnly);
(void)(curMetric);
std::vector<Field> nextRequiredFields;
if (nextMetric != "") {
bool canBeComputed = CanThisMetricBeComputed(nextMetric, hdfBasReader, hdfPlsReader,
useBaseFile, usePulseFile, nextRequiredFields);
assert(canBeComputed);
}
for (size_t i = 0; i < cachedFields.size(); i++) {
bool isRequiredForNextMetric = false;
for (size_t j = 0; j < nextRequiredFields.size(); j++) {
if (cachedFields[i] == nextRequiredFields[j]) {
isRequiredForNextMetric = true;
break;
}
}
if (isRequiredForNextMetric) {
continue;
}
std::string &curField = cachedFields[i].name;
FieldType &fieldType = cachedFields[i].type;
if (fieldType == BasField and useBaseFile and hdfBasReader.FieldIsIncluded(curField) and
hdfBasReader.includedFields[curField]) {
hdfBasReader.ClearField(baseFile, curField);
// Remove it from cachedFields
cachedFields.erase(cachedFields.begin() + i);
i--;
} else if (fieldType == PlsField and usePulseFile and
hdfPlsReader.FieldIsIncluded(curField) and
hdfPlsReader.includedFields[curField]) {
if (curField == "NumEvent") {
// Always keep NumEvent
continue;
}
hdfPlsReader.ClearField(pulseFile, curField);
// Remove it from cachedFields
cachedFields.erase(cachedFields.begin() + i);
i--;
}
}
}
// Compute StartFrame from BaseCalls only.
// Return true if succeed, false otherwise.
bool ComputeStartFrameFromBase(BaseFile &baseFile, HDFBasReader &hdfBasReader,
const bool &useBaseFile, MovieAlnIndexLookupTable &lookupTable,
std::vector<UInt> &newStartFrame)
{
newStartFrame.resize(lookupTable.readLength);
if (useBaseFile and hdfBasReader.FieldIsIncluded("PreBaseFrames") and
hdfBasReader.includedFields["PreBaseFrames"] and baseFile.preBaseFrames.size() > 0) {
// baseFile.preBaseFrame data type = uint16
// startFrame data type = uint32
for (int i = 0; i < lookupTable.readLength; i++) {
newStartFrame[i] = baseFile.preBaseFrames[lookupTable.readStart + i];
}
for (int i = 0; i < lookupTable.readLength - 1; i++) {
newStartFrame[i + 1] += baseFile.basWidthInFrames[lookupTable.readStart + i];
}
std::partial_sum(&newStartFrame[0], &newStartFrame[lookupTable.readLength],
&newStartFrame[0]);
return true;
}
return false;
}
// Compute StartFrame from PulseCalls only.
// Return true if succeed, false otherwise.
bool ComputeStartFrameFromPulse(PulseFile &pulseFile, HDFPlsReader &hdfPlsReader,
const bool &usePulseFile, MovieAlnIndexLookupTable &lookupTable,
std::vector<int> &baseToPulseIndexMap,
std::vector<UInt> &newStartFrame)
{
newStartFrame.resize(lookupTable.readLength);
if (usePulseFile) {
assert(pulseFile.startFrame.size() > 0);
hdfPlsReader.CopyFieldAt(pulseFile, "StartFrame", lookupTable.plsReadIndex,
&baseToPulseIndexMap[0], &newStartFrame[0],
lookupTable.readLength);
return true;
}
return false;
}
// Compute StartFrame from either (1) BaseCalls or (2) PulseCalls.
// (1) Uses baseFile.preBaseFrames and baseFile.basWidthInFrames
// (2) Uses pulseFile.startFrame
// In theory, the generated results using both methods should
// be exactly the same. However, they can be different in practice
// because PreBaseFrames is of data type uint_16, while its
// value can exceed maximum uint_16 (65535).
// When possible, always use PulseCalls.
void ComputeStartFrame(BaseFile &baseFile, PulseFile &pulseFile, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, bool useBaseFile, bool usePulseFile,
MovieAlnIndexLookupTable &lookupTable, std::vector<int> &baseToPulseIndexMap,
std::vector<UInt> &newStartFrame)
{
if (!ComputeStartFrameFromPulse(pulseFile, hdfPlsReader, usePulseFile, lookupTable,
baseToPulseIndexMap, newStartFrame)) {
if (!ComputeStartFrameFromBase(baseFile, hdfBasReader, useBaseFile, lookupTable,
newStartFrame)) {
std::cout << "ERROR! There is insufficient data to compute metric: StartFrame."
<< std::endl;
std::exit(EXIT_FAILURE);
}
}
}
//
// Compute and write an entire metric to cmp.h5.
// Assume that all required fields have been loaded.
//
void WriteMetric(CmpFile &cmpFile, BaseFile &baseFile, PulseFile &pulseFile,
HDFCmpFile<CmpAlignment> &cmpReader, HDFBasReader &hdfBasReader,
HDFPlsReader &hdfPlsReader, HDFCCSReader<SMRTSequence> &hdfCcsReader,
const bool &useBaseFile, const bool &usePulseFile, const bool &useCcsOnly,
std::vector<MovieAlnIndexLookupTable> &lookupTables,
std::vector<std::pair<UInt, UInt> > &groupedLookupTablesIndexPairs,
const std::string &curMetric)
{
(void)(cmpFile);
(void)(hdfCcsReader);
(void)(useCcsOnly);
for (size_t index = 0; index < groupedLookupTablesIndexPairs.size(); index++) {
// Group[index] contains all items in lookupTables[firstIndex...lastIndex)
UInt firstIndex = groupedLookupTablesIndexPairs[index].first;
UInt lastIndex = groupedLookupTablesIndexPairs[index].second;
assert(lookupTables.size() > firstIndex);
UInt refGroupIndex = lookupTables[firstIndex].refGroupIndex;
UInt readGroupIndex = lookupTables[firstIndex].readGroupIndex;
// Obtain alignment array length from *.cmp.h5/refGroup/readGroup/AlnArray.
HDFCmpExperimentGroup *expGroup =
cmpReader.refAlignGroups[refGroupIndex]->readGroups[readGroupIndex];
UInt alnArrayLength = expGroup->alignmentArray.size();
//
// Compute any necessary data fields. These usually involve
// using differences of pulse indices, pulse widths, etc..
// Missing fields are stored as 0's.
//
std::vector<UInt> startTimeOffsetMetric;
// pulseIndex's data type is uint16 in ICD,
// but I have seen it defined as uint32 in a bas file.
std::vector<UInt> pulseMetric;
std::vector<UChar> qvMetric;
std::vector<HalfWord> frameRateMetric;
std::vector<UInt> timeMetric;
std::vector<char> tagMetric;
std::vector<float> floatMetric;
/*
if (curMetric == "StartTimeOffset") {
startTimeOffsetMetric.resize(alnNum);
HDFArray<UInt> * data = (HDFArray<UInt>*) expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnNum);
data->UpdateH5Dataspace();
data->Read(0, alnNum-1, &StartTimeOffsetMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
std::fill(startTimeOffsetMetric.begin(), startTimeOffsetMetric.end(), );
}
} else */
if (curMetric == "QualityValue" || curMetric == "InsertionQV" ||
curMetric == "DeletionQV" || curMetric == "MergeQV" || curMetric == "SubstitutionQV") {
qvMetric.resize(alnArrayLength);
HDFArray<UChar> *data = (HDFArray<UChar> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &qvMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
}
} else if (curMetric == "ClassifierQV" || curMetric == "pkmid") {
// Note that data type of pkmid=midSignal, which is uint_8 in bas/pls files,
// has been changed to float in cmp.h5. Why?
floatMetric.resize(alnArrayLength);
HDFArray<float> *data = (HDFArray<float> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &floatMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(floatMetric.begin(), floatMetric.end(), NaN);
}
} else if (curMetric == "PulseIndex") {
pulseMetric.resize(alnArrayLength);
HDFArray<UInt> *data = (HDFArray<UInt> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &pulseMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(pulseMetric.begin(), pulseMetric.end(), 0);
}
} else if (curMetric == "DeletionTag" || curMetric == "SubstitutionTag") {
tagMetric.resize(alnArrayLength);
HDFArray<char> *data = (HDFArray<char> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &tagMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(tagMetric.begin(), tagMetric.end(), '-');
}
} else if (curMetric == "StartFrame" || curMetric == "StartFrameBase" ||
curMetric == "StartFramePulse") {
timeMetric.resize(alnArrayLength);
HDFArray<UInt> *data = (HDFArray<UInt> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &timeMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(timeMetric.begin(), timeMetric.end(), missingPulseIndex);
}
} else if (curMetric == "PulseWidth" || curMetric == "PreBaseFrames" ||
curMetric == "WidthInFrames" || curMetric == "IPD" || curMetric == "Light") {
frameRateMetric.resize(alnArrayLength);
HDFArray<HalfWord> *data = (HDFArray<HalfWord> *)expGroup->fields[curMetric];
if (data->IsInitialized()) {
assert(data->size() == alnArrayLength);
data->UpdateH5Dataspace();
data->Read(0, alnArrayLength - 1, &frameRateMetric[0]);
} else {
data->Initialize(expGroup->experimentGroup, curMetric);
//std::fill(frameRateMetric.begin(), frameRateMetric.end(), missingFrameRateValue);
}
} else {
std::cout << "ERROR, metric " << curMetric << " is not supported." << std::endl;
std::exit(EXIT_FAILURE);
}
for (size_t movieAlignmentIndex = firstIndex; movieAlignmentIndex < lastIndex;
movieAlignmentIndex++) {
MovieAlnIndexLookupTable &lookupTable = lookupTables[movieAlignmentIndex];
if (lookupTable.skip) continue;
const UInt alignedSequenceLength = lookupTable.offsetEnd - lookupTable.offsetBegin;
const UInt ungappedAlignedSequenceLength =
lookupTable.queryEnd - lookupTable.queryStart;
const UInt &plsReadIndex = lookupTable.plsReadIndex;
const UInt &readStart = lookupTable.readStart;
const UInt &readLength = lookupTable.readLength;
const UInt &queryStart = lookupTable.queryStart;
const UInt &offsetBegin = lookupTable.offsetBegin;
const UInt &offsetEnd = lookupTable.offsetEnd;
assert(offsetEnd <= alnArrayLength);
assert(offsetBegin + alignedSequenceLength <= alnArrayLength);
// Condense gaps and get ungapped aligned sequence.
std::string ungappedAlignedSequence = lookupTable.alignedSequence;
RemoveGaps(ungappedAlignedSequence, ungappedAlignedSequence);
std::vector<int> baseToAlignmentMap;
// Map bases in the aligned sequence to their positions in the alignment.
CreateSequenceToAlignmentMap(lookupTable.alignedSequence, baseToAlignmentMap);
std::vector<int> baseToPulseIndexMap;
if (usePulseFile && IsPulseMetric(curMetric)) {
// Map bases in the read to pulse indices.
MapBaseToPulseIndex(baseFile, pulseFile, lookupTable, baseToPulseIndexMap);
}
UInt i;
if (curMetric == "QualityValue") {
assert(baseFile.qualityValues.size() > 0 &&
baseFile.qualityValues.size() >= readStart + readLength);
std::fill(&qvMetric[offsetBegin], &qvMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
// cap quality value
qvMetric[offsetBegin + baseToAlignmentMap[i]] = std::min(
maxQualityValue, baseFile.qualityValues[readStart + queryStart + i]);
}
qvMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "InsertionQV") {
assert(baseFile.insertionQV.size() > 0 &&
baseFile.insertionQV.size() >= readStart + readLength);
std::fill(&qvMetric[offsetBegin], &qvMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
// cap quality value
qvMetric[offsetBegin + baseToAlignmentMap[i]] =
std::min(maxQualityValue, baseFile.insertionQV[readStart + queryStart + i]);
}
qvMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "MergeQV") {
assert(baseFile.mergeQV.size() > 0 &&
baseFile.mergeQV.size() >= readStart + readLength);
std::fill(&qvMetric[offsetBegin], &qvMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
// cap quality value
qvMetric[offsetBegin + baseToAlignmentMap[i]] =
std::min(maxQualityValue, baseFile.mergeQV[readStart + queryStart + i]);
}
qvMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "DeletionQV") {
assert(baseFile.deletionQV.size() > 0 &&
baseFile.deletionQV.size() >= readStart + readLength);
std::fill(&qvMetric[offsetBegin], &qvMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
// cap quality value
qvMetric[offsetBegin + baseToAlignmentMap[i]] =
std::min(maxQualityValue, baseFile.deletionQV[readStart + queryStart + i]);
}
qvMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "DeletionTag") {
assert(baseFile.deletionTag.size() > 0 &&
baseFile.deletionTag.size() >= readStart + readLength);
std::fill(&tagMetric[offsetBegin], &tagMetric[offsetEnd], '-');
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
assert(offsetBegin + baseToAlignmentMap[i] < tagMetric.size());
tagMetric[offsetBegin + baseToAlignmentMap[i]] =
baseFile.deletionTag[readStart + queryStart + i];
}
tagMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "PulseIndex") {
assert(baseFile.pulseIndex.size() > 0 &&
baseFile.pulseIndex.size() >= readStart + readLength);
std::fill(&pulseMetric[offsetBegin], &pulseMetric[offsetEnd], 0);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
pulseMetric[offsetBegin + baseToAlignmentMap[i]] =
baseFile.pulseIndex[readStart + queryStart + i];
}
pulseMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "SubstitutionTag") {
assert(baseFile.substitutionTag.size() > 0 &&
baseFile.substitutionTag.size() >= readStart + readLength);
std::fill(&tagMetric[offsetBegin], &tagMetric[offsetEnd], '-');
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
tagMetric[offsetBegin + baseToAlignmentMap[i]] =
baseFile.substitutionTag[readStart + queryStart + i];
}
tagMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "SubstitutionQV") {
assert(baseFile.substitutionQV.size() > 0 &&
baseFile.substitutionQV.size() >= readStart + readLength);
std::fill(&qvMetric[offsetBegin], &qvMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[offsetBegin + baseToAlignmentMap[i]] = std::min(
maxQualityValue, baseFile.substitutionQV[readStart + queryStart + i]);
}
qvMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "ClassifierQV") {
assert(pulseFile.classifierQV.size() > 0 &&
pulseFile.classifierQV.size() >= readStart + readLength);
std::vector<float> newClassifierQV;
newClassifierQV.resize(ungappedAlignedSequenceLength);
// For the data used for this table, it is possible to simply
// reference the data for the bas file, but for the pls file,
// it is necessary to copy since there is a packing of data.
hdfPlsReader.CopyFieldAt(pulseFile, "ClassifierQV", plsReadIndex,
&baseToPulseIndexMap[queryStart], &newClassifierQV[0],
ungappedAlignedSequenceLength);
std::fill(&floatMetric[offsetBegin], &floatMetric[offsetEnd], NaN);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
floatMetric[offsetBegin + baseToAlignmentMap[i]] = newClassifierQV[i];
}
floatMetric[offsetBegin + alignedSequenceLength] = 0;
/* } else if (curMetric == "StartTimeOffset") {
// StartTimeOffset is a subset of StartFrame.
std::vector<UInt> newStartFrame;
ComputeStartFrame(baseFile, pulseFile, hdfBasReader, hdfPlsReader,
useBaseFile, usePulseFile, lookupTable,
baseToPulseIndexMap, newStartFrame);
startTimeOffsetMetric[offsetBegin] = newStartFrame[queryStart];
*/
} else if (curMetric == "StartFrame") {
std::vector<UInt> newStartFrame;
ComputeStartFrame(baseFile, pulseFile, hdfBasReader, hdfPlsReader, useBaseFile,
usePulseFile, lookupTable, baseToPulseIndexMap, newStartFrame);
std::fill(&timeMetric[offsetBegin], &timeMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
timeMetric[offsetBegin + baseToAlignmentMap[i]] = newStartFrame[queryStart + i];
}
timeMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "StartFrameBase") {
// Sneaky metric, compute StartFrame from BaseCalls only.
std::vector<UInt> newStartFrame;
ComputeStartFrameFromBase(baseFile, hdfBasReader, useBaseFile, lookupTable,
newStartFrame);
std::fill(&timeMetric[offsetBegin], &timeMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
timeMetric[offsetBegin + baseToAlignmentMap[i]] = newStartFrame[queryStart + i];
}
timeMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "StartFramePulse") {
// Sneaky metric, compute StartFrame from PulseCalls only.
std::vector<UInt> newStartFrame;
ComputeStartFrameFromPulse(pulseFile, hdfPlsReader, usePulseFile, lookupTable,
baseToPulseIndexMap, newStartFrame);
std::fill(&timeMetric[offsetBegin], &timeMetric[offsetEnd], missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
timeMetric[offsetBegin + baseToAlignmentMap[i]] = newStartFrame[queryStart + i];
}
timeMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "PreBaseFrames") {
// Directly load baseFile.PreBaseFrames.
// DON'T compute it from PulseCalls even if you can.
assert(baseFile.preBaseFrames.size() > 0 &&
baseFile.preBaseFrames.size() >= readStart + readLength);
std::fill(&frameRateMetric[offsetBegin], &frameRateMetric[offsetEnd],
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] =
baseFile.preBaseFrames[readStart + queryStart + i];
}
frameRateMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "WidthInFrames" || curMetric == "PulseWidth") {
// For legacy reasons, it's possible the width in frames is
// stored in the bas file. If this is the case, use the width
// in frames there. Otherwise, use the width in frames stored
// in the pls file.
std::vector<uint16_t> newWidthInFrames;
newWidthInFrames.resize(ungappedAlignedSequenceLength);
if (usePulseFile) {
hdfPlsReader.CopyFieldAt(pulseFile, "WidthInFrames", plsReadIndex,
&baseToPulseIndexMap[queryStart], &newWidthInFrames[0],
ungappedAlignedSequenceLength);
} else if (useBaseFile) {
// basWidthInFrames data type uint16
std::copy(&baseFile.basWidthInFrames[readStart + queryStart],
&baseFile.basWidthInFrames[readStart + queryStart +
ungappedAlignedSequenceLength],
&newWidthInFrames[0]);
}
std::fill(&frameRateMetric[offsetBegin], &frameRateMetric[offsetEnd],
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] = newWidthInFrames[i];
}
frameRateMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "pkmid") {
// pkmid in cmp.h5 is MidSignal in pls.h5, but
// data type of MidSignal is uint16 in pls files,
// data type of pkmid is float in cmp files.
assert(usePulseFile);
std::vector<HalfWord> newMidSignal;
newMidSignal.resize(ungappedAlignedSequenceLength);
hdfPlsReader.CopyFieldAt(pulseFile, "MidSignal", plsReadIndex,
&baseToPulseIndexMap[queryStart], &newMidSignal[0],
ungappedAlignedSequenceLength, ungappedAlignedSequence);
std::fill(&floatMetric[offsetBegin], &floatMetric[offsetEnd], NaN);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
floatMetric[offsetBegin + baseToAlignmentMap[i]] = newMidSignal[i];
}
floatMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "IPD") {
std::fill(&frameRateMetric[offsetBegin], &frameRateMetric[offsetEnd],
missingFrameRateValue);
// IPD can be either (1) copied from baseFile.preBaseFrames
// or (2) computed from pulseFile.StartFrame and pulseFile.WidthInFrames
// Always use method (2) when possible as it is more accurate.
if (usePulseFile) {
// Need to read StartFrame & WidthInFrames for the entire read,
// not only for a subset of bases in the alignment
assert(pulseFile.startFrame.size() > 0);
assert(pulseFile.plsWidthInFrames.size() > 0);
std::vector<UInt> newStartFrame;
newStartFrame.resize(readLength);
hdfPlsReader.CopyFieldAt(pulseFile, "StartFrame", plsReadIndex,
&baseToPulseIndexMap[0], &newStartFrame[0],
readLength);
std::vector<uint16_t> newWidthInFrames;
newWidthInFrames.resize(readLength);
hdfPlsReader.CopyFieldAt(pulseFile, "WidthInFrames", plsReadIndex,
&baseToPulseIndexMap[0], &newWidthInFrames[0],
readLength);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
// The IPD is undefined for the first base in a read.
if (queryStart == 0 and i == 0) {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] = 0;
} else {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] =
newStartFrame[queryStart + i] - newStartFrame[i + queryStart - 1] -
newWidthInFrames[i + queryStart - 1];
}
}
} else if (useBaseFile) {
assert(baseFile.preBaseFrames.size() > 0);
assert(baseFile.preBaseFrames.size() >= readStart + readLength);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] =
baseFile.preBaseFrames[readStart + queryStart + i];
}
}
frameRateMetric[offsetBegin + alignedSequenceLength] = 0;
} else if (curMetric == "Light") {
// Light can be computed from pulseFile.meanSignal and
// pulseFile.plsWidthInFrames. Might have been deprecated.
assert(usePulseFile);
std::fill(&frameRateMetric[offsetBegin], &frameRateMetric[offsetEnd],
missingFrameRateValue);
std::vector<uint16_t> newMeanSignal;
newMeanSignal.resize(ungappedAlignedSequenceLength);
hdfPlsReader.CopyFieldAt(pulseFile, "MeanSignal", plsReadIndex,
&baseToPulseIndexMap[queryStart], &newMeanSignal[0],
ungappedAlignedSequenceLength, ungappedAlignedSequence);
std::vector<uint16_t> newWidthInFrames;
newWidthInFrames.resize(ungappedAlignedSequenceLength);
hdfPlsReader.CopyFieldAt(pulseFile, "WidthInFrames", plsReadIndex,
&baseToPulseIndexMap[queryStart], &newWidthInFrames[0],
ungappedAlignedSequenceLength);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[offsetBegin + baseToAlignmentMap[i]] =
newMeanSignal[i] * newWidthInFrames[i];
}
frameRateMetric[offsetBegin + alignedSequenceLength] = 0;
} else {
std::cout << "ERROR, unknown metric " << curMetric << std::endl;
std::exit(EXIT_FAILURE);
}
}
// Write the computed metric to cmp.h5.
/*if (curMetric == "StartTimeOffset") {
expGroup->startTimeOffset.WriteToPos(&startTimeOffsetMetric[0], startTimeOffsetMetric.size(), 0);
} else */
if (curMetric == "QualityValue" || curMetric == "InsertionQV" ||
curMetric == "DeletionQV" || curMetric == "MergeQV" || curMetric == "SubstitutionQV") {
HDFArray<UChar> *data = (HDFArray<UChar> *)expGroup->fields[curMetric];
data->WriteToPos(&qvMetric[0], qvMetric.size(), 0);
} else if (curMetric == "ClassifierQV" || curMetric == "pkmid") {
HDFArray<float> *data = (HDFArray<float> *)expGroup->fields[curMetric];
data->WriteToPos(&floatMetric[0], floatMetric.size(), 0);
} else if (curMetric == "PulseIndex") {
HDFArray<UInt> *data = (HDFArray<UInt> *)expGroup->fields[curMetric];
data->WriteToPos(&pulseMetric[0], pulseMetric.size(), 0);
} else if (curMetric == "DeletionTag" || curMetric == "SubstitutionTag") {
HDFArray<char> *data = (HDFArray<char> *)expGroup->fields[curMetric];
data->WriteToPos(&tagMetric[0], tagMetric.size(), 0);
} else if (curMetric == "StartFrame" || curMetric == "StartFrameBase" ||
curMetric == "StartFramePulse") {
HDFArray<UInt> *data = (HDFArray<UInt> *)expGroup->fields[curMetric];
data->WriteToPos(&timeMetric[0], timeMetric.size(), 0);
} else if (curMetric == "PulseWidth" || curMetric == "PreBaseFrames" ||
curMetric == "WidthInFrames" || curMetric == "IPD" || curMetric == "Light") {
HDFArray<HalfWord> *data = (HDFArray<HalfWord> *)expGroup->fields[curMetric];
data->WriteToPos(&frameRateMetric[0], frameRateMetric.size(), 0);
} else {
std::cout << "ERROR, unknown metric " << curMetric << std::endl;
std::exit(EXIT_FAILURE);
}
}
}
//
// Write "WhenStarted" from pls.h5 and write to cmp.h5
//
void WriteMetricWhenStarted(HDFCmpFile<CmpAlignment> &cmpReader, HDFPlsReader &hdfPlsReader,
const std::string &movieName)
{
std::string metric = "WhenStarted";
std::string whenStarted;
if (hdfPlsReader.scanDataReader.useWhenStarted == false) {
std::cout << "ERROR! Attempting to read WhenStarted from " << movieName
<< " but the attriubte does not exist." << std::endl;
std::exit(EXIT_FAILURE);
}
hdfPlsReader.scanDataReader.ReadWhenStarted(whenStarted);
if (!cmpReader.movieInfoGroup.whenStartedArray.IsInitialized()) {
cmpReader.movieInfoGroup.whenStartedArray.Initialize(
cmpReader.movieInfoGroup.movieInfoGroup, metric);
}
cmpReader.movieInfoGroup.whenStartedArray.Write(&whenStarted, 1);
}
//
// Print metrics.
//
std::string MetricsToString(const std::vector<std::string> &metrics)
{
std::string ret = "";
for (size_t i = 0; i < metrics.size(); i++) {
ret += metrics[i];
if (i != metrics.size() - 1) ret += ",";
if (i % 4 == 3) ret += "\n";
}
return ret;
}
//
// Print usage.
//
void PrintUsage()
{
std::cout << " loadPulses - Load pulse information and quality values into a Compare file"
<< std::endl;
std::cout << "usage: loadPulses movieFile cmpFile [-metrics m1,m2,...] [-byread]" << std::endl;
std::cout << " movieFile may be a movie file or a fofn of movie file names." << std::endl;
std::cout << " metrics m1,m2,... is a comma-separated list (without spaces) of metrics "
<< std::endl
<< " to print to the pulse file." << std::endl;
std::cout << " Valid metrics are: " << std::endl;
std::cout << MetricsToString(GetAllSupportedMetrics(false)) << std::endl;
// << " QualityValue, ClassifierQV, MergeQV," << std::endl
// << " StartFrame, PulseWidth, pkmid, IPD, Light" << std::endl
// << " WhenStarted, StartTimeOffset, PreBaseFrames," << std::endl
// << " InsertionQV, DeletionQV, DeletionTag, SubstitutionQV" << std::endl
// << " SubstitutionTag, PulseIndex, WidthInFrames" << std::endl;
std::cout << " By default, " << MetricsToString(GetDefaultMetrics()) << " are added"
<< std::endl;
// Deprecate -useccs, an option for old data.
// std::cout << " -useccs This option is for older cmp.h5 files that do not have the read type " << std::endl
// << " stored. Newer cmp.h5 files have a read type that indicates the cmp.h5 file " << std::endl
// << " has alignments generated from de novo ccs sequences. Using this flag assuems"<<std::endl
// << " ALL alignments in the cmp.h5 file are from ccs sequences, and loads the "<< std::endl
// << " quality values from ccs instead of the raw sequence. "<<std::endl
// << " The only metrics that are allowed for de novo ccs sequences are QualityValue, " << std::endl
// << " InsertionQV, DeletionQV, and SubstitutionQV" << std::endl;
std::cout << " -byread Reads pulse/base fields by read, rather than reading an entire "
<< std::endl
<< " movie first. This uses considerably less memory than the defualt mode"
<< std::endl
<< " but is slow." << std::endl;
std::cout << " -byMetric Loads every pls/base field for each movie entirely before loading "
<< std::endl
<< " another field. This uses more memory than -byread, but can be faster."
<< std::endl
<< " This opiton is experimental. " << std::endl;
std::cout << " Using hdf version " << H5_VERS_MAJOR << "." << H5_VERS_MINOR << "."
<< H5_VERS_RELEASE << std::endl;
}
//
// The main function.
//
int main(int argc, char *argv[])
{
std::string program = "loadPulses";
std::string versionStr(VERSION);
AppendPerforceChangelist(PERFORCE_VERSION_STRING, versionStr);
std::string cmpFileName, movieFileName;
std::map<std::string, bool> metricOptions;
int maxElements = 0;
//Maximum Memory allowed for bymetric is 6 GB
int maxMemory = 4;
//
// Default is all options are false
//
CreateMetricOptions(metricOptions);
std::string metricList = "";
bool useCcsOnly = false;
bool byRead = false;
bool byMetric = false;
bool failOnMissingData = false;
CommandLineParser clp;
clp.SetProgramName(program);
clp.SetVersion(versionStr);
clp.RegisterStringOption("basFileName", &movieFileName, "The input {bas,pls}.h5 or input.fofn.",
true);
clp.RegisterStringOption("cmpFileName", &cmpFileName,
"The cmp.h5 file to load pulse information into.", true);
clp.RegisterPreviousFlagsAsHidden();
std::string metricsDescription =
"A comma separated list of metrics (with no spaces).\nValid options are:\n";
metricsDescription += MetricsToString(GetAllSupportedMetrics(false));
metricsDescription += "\nDefault options are:\n";
metricsDescription += MetricsToString(GetDefaultMetrics());
clp.RegisterStringOption("metrics", &metricList, metricsDescription);
clp.RegisterFlagOption("failOnMissingData", &failOnMissingData,
"Exit if any data fields are missing from the bas.h5 or pls.h5 "
"input that are required to load a metric. Defualt is a warning.");
clp.RegisterFlagOption("byread", &byRead,
"Load pulse information by read rather than buffering metrics.");
clp.RegisterFlagOption("bymetric", &byMetric,
"Load pulse information by metric rather than by read. "
"This uses more memory than -byread, but can be faster.");
clp.RegisterIntOption("maxElements", &maxElements,
"Set a limit on the size of pls/bas file to buffer in with -bymetric "
"(default value: maximum int). Use -byread if the limit is exceeded.",
CommandLineParser::PositiveInteger);
clp.RegisterIntOption("maxMemory", &maxMemory,
"Set a limit (in GB) on the memory to buffer data with -bymetric "
"(default value: 4 GB). Use -byread if the limit is exceeded.",
CommandLineParser::PositiveInteger);
int metaNElements, rawChunkSize, rawNElements;
metaNElements = 0;
rawChunkSize = 0;
metaNElements = 0;
clp.RegisterIntOption(
"metaNElements", &metaNElements,
"Set number of elements in meta data cache for reading bas/bax/pls.h5 file.",
CommandLineParser::PositiveInteger);
clp.RegisterIntOption(
"rawNElements", &rawNElements,
"Set number of elements in raw data cache for reading bas/bax/pls.h5 file.",
CommandLineParser::PositiveInteger);
clp.RegisterIntOption("rawChunkSize", &rawChunkSize,
"Set chunk size of raw data cache for reading bas/bax/pls.h5 file.",
CommandLineParser::PositiveInteger);
std::string progSummary =
("Loads pulse information such as inter pulse "
"distance, or quality information into the cmp.h5 file. This allows "
"one to analyze kinetic and quality information by alignment column.");
clp.SetProgramSummary(progSummary);
clp.ParseCommandLine(argc, argv);
std::cerr << "[INFO] " << GetTimestamp() << " [" << program << "] started." << std::endl;
//use byMetric by default unless byRead is specified.
byMetric = true;
if (byRead) {
byMetric = false;
}
if (metricList == "") {
SetDefaultMetricOptions(metricOptions);
} else {
ParseMetricsList(metricList, metricOptions);
}
//
// Always read in basecalls since they are used to check the sanity
// of the alignment indices.
//
metricOptions["Basecall"] = true;
//
// Translate from the metrics to be loaded to the ones that are
// required to compute them.
// Need to be refactored.
//
std::vector<std::string> datasetFields;
RequirementMap fieldRequirements;
BuildRequirementMap(fieldRequirements);
StoreDatasetFieldsFromPulseFields(metricOptions, fieldRequirements, datasetFields);
//e.g. /PATH_TO_FILE/m120321_032600_42142_c100310572550000001523013208061210_s1_p0.bas.h5
// /PATH_TO_FILE/m120321_032600_42142_c100310572550000001523013208061210_s2_p0.bas.h5
std::vector<std::string> movieFileNames;
//e.g. m120321_032600_42142_c100310572550000001523013208061210_s1_p0
// m120321_032600_42142_c100310572550000001523013208061210_s2_p0
std::vector<std::string> fofnMovieNames;
FileOfFileNames::StoreFileOrFileList(movieFileName, movieFileNames);
HDFBasReader hdfBasReader;
HDFPlsReader hdfPlsReader;
HDFCCSReader<SMRTSequence> hdfCcsReader;
std::vector<std::string> baseFileFields, pulseFileFields;
size_t fieldIndex;
bool useBaseFile = false, usePulseFile = false;
for (fieldIndex = 0; fieldIndex < datasetFields.size(); fieldIndex++) {
if (hdfBasReader.ContainsField(datasetFields[fieldIndex])) {
useBaseFile = true;
baseFileFields.push_back(datasetFields[fieldIndex]);
}
}
if (maxElements != 0) {
hdfBasReader.maxAllocNElements = maxElements;
hdfPlsReader.maxAllocNElements = maxElements;
}
//
// For now, all runs will attempt to use information from a .bas
// file, since it's assumed that if one has alignments, one has a
// .bas file.
//
useBaseFile = true;
//
// Add some default fields.
//
hdfBasReader.IncludeField("Basecall");
hdfBasReader.IncludeField("PulseIndex");
hdfBasReader.InitializeFields(baseFileFields);
for (fieldIndex = 0; fieldIndex < datasetFields.size(); fieldIndex++) {
if (hdfPlsReader.ContainsField(datasetFields[fieldIndex])) {
usePulseFile = true;
pulseFileFields.push_back(datasetFields[fieldIndex]);
}
}
if (usePulseFile) {
// set hdfPlsReader.includedFields[fieldX] to true if fieldX is
// in pulseFileFields
hdfPlsReader.InitializeFields(pulseFileFields);
}
hdfPlsReader.IncludeField("NumEvent");
int nMovies = movieFileNames.size();
int movieIndex;
MovieNameToArrayIndex movieNameMap;
//
// Initialize movies. This accomplishes two tasks. First, all movie
// files are opened and initialized, so that if there are data
// fields missing the program will exit now rather than in the
// middle of loading pulses.
// Next, a list of movie names is created in fofnMovieNames. The
// cmp file does not necessarily index movies in the order of the
// fofn, and so when loading pulses from a movie indexed by a cmp
// file, one needs to look up the file name of the movie. This is
// done by scanning the fofnMovieNames list in order until the movie
// is found.
//
// h5 file access property list can be customized here.
//
H5::FileAccPropList fileAccPropList = H5::FileAccPropList::DEFAULT;
// h5: number of items in meta data cache
int mdc_nelmts = (metaNElements == 0) ? (4096) : (metaNElements);
// h5: number of items in raw data chunk cache
size_t rdcc_nelmts = (rawNElements == 0) ? (4096) : (rawNElements);
// h5: raw data chunk cache size (in bytes) per dataset
size_t rdcc_nbytes = (rawChunkSize == 0) ? (9192) : (rawChunkSize);
double rdcc_w0 = 0.75; // h5: preemption policy
// fileAccPropList.getCache(mdc_nelmts, rdcc_nelmts, rdcc_nbytes, rdcc_w0);
fileAccPropList.setCache(mdc_nelmts, rdcc_nelmts, rdcc_nbytes, rdcc_w0);
// fileAccPropList.setCache(4096, 4096, 8388608, rdcc_w0);
// If one of the h5 in the fofn is a ccs.h5 file, then only load pulse
// information from group /PulseData/ConsensusBaseCalls.
for (movieIndex = 0; movieIndex < nMovies; movieIndex++) {
FileType fileType;
BaseSequenceIO::DetermineFileTypeByExtension(movieFileNames[movieIndex], fileType, true);
if (fileType == FileType::HDFCCSONLY) {
useCcsOnly = true;
}
}
for (movieIndex = 0; movieIndex < nMovies; movieIndex++) {
if (useCcsOnly) {
hdfCcsReader.SetReadBasesFromCCS();
hdfBasReader.SetReadBasesFromCCS();
}
if (!hdfBasReader.Initialize(movieFileNames[movieIndex], fileAccPropList)) {
std::cout << "ERROR, could not initialize HDF file " << movieFileNames[movieIndex]
<< " for reading bases." << std::endl;
std::exit(EXIT_FAILURE);
} else {
fofnMovieNames.push_back(hdfBasReader.GetMovieName());
movieNameMap[hdfBasReader.GetMovieName()] = movieIndex;
hdfBasReader.Close();
}
//
// The pulse file is optional.
//
if (usePulseFile) {
if (hdfPlsReader.Initialize(movieFileNames[movieIndex], fileAccPropList) == 0) {
usePulseFile = false;
}
}
}
CmpFile cmpFile;
//
// These readers pull information from the same pls file.
//
HDFCmpFile<CmpAlignment> cmpReader;
if (cmpReader.Initialize(cmpFileName, H5F_ACC_RDWR) == 0) {
std::cout << "ERROR, could not open the cmp file." << std::endl;
std::exit(EXIT_FAILURE);
}
if (cmpReader.HasNoAlignments()) {
std::cout << "WARNING, there is no alignment in the cmp file." << std::endl;
if (useBaseFile) {
hdfBasReader.Close();
}
if (usePulseFile) {
hdfPlsReader.Close();
}
cmpReader.Close();
std::cerr << "[INFO] " << GetTimestamp() << " [" << program << "] ended." << std::endl;
std::exit(EXIT_SUCCESS);
}
cmpReader.Read(cmpFile, false);
// Sanity check: if there is a ccs.h5 file in the fofn and
// cmp.h5 file's readType is not CCS, something is wrong.
if (cmpFile.readType != ReadType::CCS and useCcsOnly) {
std::cout << "ERROR, there is a ccs.h5 file in the fofn, while read type of"
<< " the cmp.h5 file is not CCS." << std::endl;
std::exit(EXIT_FAILURE);
}
std::string commandLine;
clp.CommandLineToString(argc, argv, commandLine);
cmpReader.fileLogGroup.AddEntry(commandLine, "Loading pulse metrics", program, GetTimestamp(),
versionStr);
//
// Group alignment indices by movie so that they may be processed one movie at a time
// later on. The movie indices set keeps track of all indices
// listed in alignment files. This keeps a reference to all
// alignments in memory at once. At the time of writing this, most
// projects will have at most a few million alignments, and so the
// size of this structure is modest.
// Each movieIndexSets[$movieId] contains indices of all the alignments, which
// are associated with a movie whose id in dataset /MovieInfo/ID equals $movieId
//
UInt alignmentIndex;
std::map<int, std::vector<int> > movieIndexSets;
for (alignmentIndex = 0; alignmentIndex < cmpFile.alnInfo.alignments.size(); alignmentIndex++) {
movieIndexSets[cmpFile.alnInfo.alignments[alignmentIndex].GetMovieId()].push_back(
alignmentIndex);
}
//
// Load pulses from movies in order they appear in the input fofn.
//
int m;
for (size_t fofnMovieIndex = 0; fofnMovieIndex < fofnMovieNames.size(); fofnMovieIndex++) {
bool byMetricForThisMovie = byMetric;
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
hdfBasReader.SetReadBasesFromCCS();
hdfCcsReader.Initialize(movieFileNames[fofnMovieIndex], fileAccPropList);
}
hdfBasReader.Initialize(movieFileNames[fofnMovieIndex], fileAccPropList);
BaseFile baseFile;
PulseFile pulseFile;
//
// Deprecate reading the entire bas.h5 file. Reads are scanned
// one by one or by metric, instead of caching all.
// It is still necessary to read in some of the datasets entirely,
// in particular the start positions and hole numbers.
//
hdfBasReader.ReadBaseFileInit(baseFile);
std::set<uint32_t> moviePartHoleNumbers;
copy(baseFile.holeNumbers.begin(), baseFile.holeNumbers.end(),
inserter(moviePartHoleNumbers, moviePartHoleNumbers.begin()));
if (usePulseFile) {
hdfPlsReader.Initialize(movieFileNames[fofnMovieIndex], fileAccPropList);
hdfPlsReader.IncludeField("NumEvent");
hdfPlsReader.IncludeField("StartFrame");
//
// Deprecate reading the entire pls.h5 file.
// Reads are scanned by read or by metric instead of caching all.
// It is still necessary to read in some of the datasets entirely,
// in particular the start positions and hole numbers.
//
hdfPlsReader.ReadPulseFileInit(pulseFile);
}
std::string cmpFileMovieName;
for (m = 0; m < static_cast<int>(cmpFile.movieInfo.name.size()); m++) {
//
// First find the file name for the movie 'm'
//
cmpFileMovieName = cmpFile.movieInfo.name[m];
if (baseFile.GetMovieName() == cmpFileMovieName) {
break;
}
}
//
// If the movie specified in the input.fofn is not found in the
// cmp file, that indicates something bad is happeing. Either the
// input.fofn was not used to generate the cmp.h5 file, or no
// alignments were found between the input bas.h5 and the
// reference. That shouldn't happen.
//
if (m == static_cast<int>(cmpFile.movieInfo.name.size())) {
std::cout << "WARNING: Could not find any alignments for file "
<< movieFileNames[fofnMovieIndex] << std::endl;
continue;
}
//
// Open the movie and load its pulses into memory.
//
movieIndex = cmpFile.movieInfo.id[m];
UInt movieAlignmentIndex;
//
// Since usePulseFile is set when the input file is a pulseFile,
// and ReadType::CCS becomes the read type when the alignments are
// ccs, when pulse files are specified for de novo ccs alignments,
// they will be opened as pulse files. Since the de novo ccs
// sequences do not have pulse file information, the auto-reading
// of pulse files needs to be disabled. Do that here.
//
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
usePulseFile = false;
}
// Check whether all metrics are computable or not.
CanMetricsBeComputed(metricOptions, hdfBasReader, hdfPlsReader, useBaseFile, usePulseFile,
failOnMissingData, movieFileNames[fofnMovieIndex]);
// Get all metrics that are (1) supported, (2) required and (3) can be loaded.
std::vector<std::string> metricsToLoad = GetMetricsToLoad(metricOptions);
//
// An index set is a set of indices into the alignment array that
// are of reads generated by this movie. Load pulses for all
// alignments generated for this movie.
//
// Movie index sets should be sorted by alignment index. Build a lookup table for this.
//
std::vector<std::pair<int, int> > toFrom;
UInt totalAlnLength = 0;
for (movieAlignmentIndex = 0; movieAlignmentIndex < movieIndexSets[movieIndex].size();
movieAlignmentIndex++) {
alignmentIndex = movieIndexSets[movieIndex][movieAlignmentIndex];
totalAlnLength += cmpFile.alnInfo.alignments[alignmentIndex].GetOffsetEnd() -
cmpFile.alnInfo.alignments[alignmentIndex].GetOffsetBegin();
toFrom.push_back(std::pair<int, int>(
cmpFile.alnInfo.alignments[alignmentIndex].GetAlignmentId(), movieAlignmentIndex));
}
// orders by first by default.
std::sort(toFrom.begin(), toFrom.end());
//
// Check metric dataset size in this movie and the required memory
// consumption, if either limit is exceeded, switch to byread.
//
if (byMetricForThisMovie) {
UInt requiredMem =
ComputeRequiredMemory(metricsToLoad, hdfBasReader, hdfPlsReader, useBaseFile,
usePulseFile, cmpReader, totalAlnLength);
if (hdfBasReader.baseArray.arrayLength >
static_cast<DSLength>(hdfBasReader.maxAllocNElements) or
(usePulseFile and
hdfPlsReader.GetStartFrameSize() > hdfPlsReader.maxAllocNElements) or
((float)requiredMem / 1024 / 1024) > maxMemory) {
std::cout << "Either the number of elements exceeds maxElement ("
<< hdfPlsReader.maxAllocNElements << "). Or the estimated memory "
<< std::endl
<< "consumption exceeds maxMemory (" << maxMemory << " GB)." << std::endl
<< "Loading pulses from " << movieFileNames[fofnMovieIndex] << " by read."
<< std::endl;
byMetricForThisMovie = false;
}
}
if (((metricOptions.find("StartFrameBase") != metricOptions.end() and
metricOptions["StartFrameBase"]) or
(metricOptions.find("StartFramePulse") != metricOptions.end() and
metricOptions["StartFramePulse"])) and
!byMetricForThisMovie) {
// Sneaky metrics StartFrameBase and StartFramePulse can used
// with -bymetric only
std::cout << "ERROR: Internal metrics StartFrameBase and StartFramePulse "
<< "can only be loaded with -bymetric." << std::endl;
std::exit(EXIT_FAILURE);
}
// Load "WhenStarted" before processing the others.
if (metricOptions["WhenStarted"]) {
WriteMetricWhenStarted(cmpReader, hdfPlsReader, movieFileNames[fofnMovieIndex]);
}
// Now load frame rate.
// if (AnyFieldRequiresFrameRate(datasetFields)) {
// Load frame rate anyway to ensure that cmp.h5 files are consistent.
if (useBaseFile) {
cmpReader.movieInfoGroup.StoreFrameRate(m, baseFile.GetFrameRate());
} else if (usePulseFile) {
cmpReader.movieInfoGroup.StoreFrameRate(m, pulseFile.GetFrameRate());
}
//
// Load metrics for alignments from movie 'movieIndex'.
//
std::cout << "loading " << movieIndexSets[movieIndex].size() << " alignments for movie "
<< movieIndex << std::endl;
if (byMetricForThisMovie) {
//
// Build lookup tables for all alignments which
// are generated by the movie and check whether
// pls/bas.h5 and cmp.h5 match.
//
std::vector<MovieAlnIndexLookupTable> lookupTables;
BuildLookupTablesAndMakeSane(cmpFile, baseFile, pulseFile, cmpReader, hdfBasReader,
hdfPlsReader, hdfCcsReader, useBaseFile, usePulseFile,
useCcsOnly, movieIndexSets[movieIndex], toFrom,
moviePartHoleNumbers, lookupTables);
//
// Group lookup tables by refGroupIndex and readGroupIndex.
//
std::vector<std::pair<UInt, UInt> > groupedLookupTablesIndexPairs;
GroupLookupTables(lookupTables, groupedLookupTablesIndexPairs);
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
std::vector<unsigned int> numPassesMetric;
numPassesMetric.resize(lookupTables.size());
UInt index = 0;
for (index = 0; index < lookupTables.size(); index++) {
if (lookupTables[index].skip) {
continue;
}
numPassesMetric[index] =
hdfCcsReader.GetNumPasses(lookupTables[index].readIndex);
}
if (!cmpReader.alnInfoGroup.numPasses.IsInitialized()) {
cmpReader.alnInfoGroup.InitializeNumPasses();
// Clear /AlnInfo/NumPasses dataset.
cmpReader.alnInfoGroup.numPasses.Resize(0);
}
// Append numPasses of this movie to the end of /AlnInfo/NumPasses.
UInt numPassesSize = cmpReader.alnInfoGroup.numPasses.size();
cmpReader.alnInfoGroup.numPasses.WriteToPos(&numPassesMetric[0],
numPassesMetric.size(), numPassesSize);
}
// Keep a list of currently cached fields.
std::vector<Field> cachedFields;
if (usePulseFile) {
// PulseCalls/ZMW/NumEvent is always cached in plsFile.
cachedFields.push_back(Field("NumEvent", PlsField));
}
for (size_t metricsToLoadIndex = 0; metricsToLoadIndex < metricsToLoad.size();
metricsToLoadIndex++) {
std::string curMetric = metricsToLoad[metricsToLoadIndex];
// Metric "WhenStarted" should have been loaded before getting here.
if (curMetric == "WhenStarted") {
continue;
}
// Get the next metric to load.
std::string nextMetric = "";
if (metricsToLoadIndex + 1 < metricsToLoad.size()) {
nextMetric = metricsToLoad[metricsToLoadIndex + 1];
}
// Cache all required data for computing this metric.
CacheRequiredFieldsForMetric(baseFile, pulseFile, hdfBasReader, hdfPlsReader,
hdfCcsReader, useBaseFile, usePulseFile, useCcsOnly,
cachedFields, curMetric);
// Compute the metric and write it to cmp.h5.
WriteMetric(cmpFile, baseFile, pulseFile, cmpReader, hdfBasReader, hdfPlsReader,
hdfCcsReader, useBaseFile, usePulseFile, useCcsOnly, lookupTables,
groupedLookupTablesIndexPairs, curMetric);
// Clear cached fields unless they are required by the next metric.
ClearCachedFields(baseFile, pulseFile, hdfBasReader, hdfPlsReader, hdfCcsReader,
useBaseFile, usePulseFile, useCcsOnly, cachedFields, curMetric,
nextMetric);
}
// Clear the default field "NumEvent"
if (usePulseFile) {
hdfPlsReader.ClearField(pulseFile, "NumEvent");
}
} else { // byRead for this movie
for (movieAlignmentIndex = 0; movieAlignmentIndex < movieIndexSets[movieIndex].size();
movieAlignmentIndex++) {
MovieAlnIndexLookupTable lookupTable;
BuildLookupTable(movieAlignmentIndex, cmpFile, baseFile, usePulseFile, pulseFile,
cmpReader, movieIndexSets[movieIndex], toFrom,
moviePartHoleNumbers, lookupTable);
// Skip this alignment if it is not generated by this movie
if (lookupTable.skip) {
continue;
}
UInt &alignmentIndex = lookupTable.alignmentIndex;
size_t &refGroupIndex = lookupTable.refGroupIndex;
size_t &readGroupIndex = lookupTable.readGroupIndex;
UInt &holeNumber = lookupTable.holeNumber;
size_t &readIndex = lookupTable.readIndex;
UInt &queryStart = lookupTable.queryStart;
UInt &queryEnd = lookupTable.queryEnd;
UInt &offsetBegin = lookupTable.offsetBegin;
UInt &offsetEnd = lookupTable.offsetEnd;
std::string alignedSequence = GetAlignedSequenceFromCmpFile(cmpReader, lookupTable);
// Create a map of where.
std::vector<int> baseToAlignmentMap;
CreateSequenceToAlignmentMap(alignedSequence, baseToAlignmentMap);
// Condense gaps in the alignment for easy comparison.
RemoveGaps(alignedSequence, alignedSequence);
// Get source read.
unsigned int numPasses;
SMRTSequence sourceRead;
GetSourceRead(cmpFile, baseFile, pulseFile, hdfBasReader, hdfPlsReader,
hdfCcsReader, useBaseFile, usePulseFile, useCcsOnly,
//byRead ,
lookupTable, alignedSequence, sourceRead, numPasses);
std::string readSequence;
readSequence.resize(queryEnd - queryStart);
copy((char *)(sourceRead.seq + queryStart), (char *)(sourceRead.seq + queryEnd),
readSequence.begin());
if (alignedSequence.size() != readSequence.size() or
alignedSequence != readSequence) {
std::cout
<< "ERROR, the query sequence does not match the aligned query sequence."
<< std::endl;
std::cout << "HoleNumber: " << holeNumber
<< ", MovieName: " << cmpFileMovieName;
std::cout << ", ReadIndex: " << (int)readIndex;
std::cout << ", qStart: " << queryStart << ", qEnd: " << queryEnd << std::endl;
std::cout << "Aligned sequence: " << std::endl;
std::cout << alignedSequence << std::endl;
std::cout << "Original sequence: " << std::endl;
std::cout << readSequence << std::endl;
assert(0);
}
//
// Compute any necessary data fields. These usually involve
// using differences of pulse indices, pulse widths, etc..
// Missing fields are stored as 0's.
//
std::vector<float> readPulseMetric;
std::vector<float> floatMetric;
std::vector<UChar> qvMetric;
std::vector<HalfWord> frameRateMetric;
std::vector<uint32_t> timeMetric;
UInt ungappedAlignedSequenceLength = alignedSequence.size();
assert(ungappedAlignedSequenceLength == queryEnd - queryStart);
UInt alignedSequenceLength = offsetEnd - offsetBegin;
readPulseMetric.resize(alignedSequenceLength + 1);
qvMetric.resize(alignedSequenceLength + 1);
frameRateMetric.resize(alignedSequenceLength + 1);
timeMetric.resize(alignedSequenceLength + 1);
UInt i;
HDFCmpExperimentGroup *expGroup =
cmpReader.refAlignGroups[refGroupIndex]->readGroups[readGroupIndex];
UInt alnArrayLength = expGroup->alignmentArray.size();
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
if (!cmpReader.alnInfoGroup.numPasses.IsInitialized()) {
cmpReader.alnInfoGroup.InitializeNumPasses();
}
cmpReader.alnInfoGroup.numPasses.WriteToPos(&numPasses, 1, alignmentIndex);
}
if (metricOptions["StartTimeOffset"] == true) {
if (!expGroup->startTimeOffset.IsInitialized()) {
expGroup->startTimeOffset.Initialize(expGroup->experimentGroup,
"StartTimeOffset");
}
unsigned int readStartTimeOffset = sourceRead.startFrame[queryStart];
expGroup->startTimeOffset.WriteToPos(&readStartTimeOffset, 1, alignmentIndex);
}
if (metricOptions["QualityValue"] == true) {
if (!expGroup->qualityValue.IsInitialized()) {
expGroup->qualityValue.Initialize(expGroup->experimentGroup, "QualityValue",
true, alnArrayLength);
}
// Store QualityValue.
std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[baseToAlignmentMap[i]] = sourceRead.qual[queryStart + i];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->qualityValue.WriteToPos(&qvMetric[0], qvMetric.size(), offsetBegin);
}
if (metricOptions["InsertionQV"] == true) {
if (!expGroup->insertionQV.IsInitialized()) {
expGroup->insertionQV.Initialize(expGroup->experimentGroup, "InsertionQV",
true, alnArrayLength);
}
// Store InsertionQV.
std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[baseToAlignmentMap[i]] = sourceRead.insertionQV[queryStart + i];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->insertionQV.WriteToPos(&qvMetric[0], qvMetric.size(), offsetBegin);
}
if (metricOptions["MergeQV"] == true) {
if (!expGroup->mergeQV.IsInitialized()) {
expGroup->mergeQV.Initialize(expGroup->experimentGroup, "MergeQV", true,
alnArrayLength);
}
// Store MergeQV.
std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[baseToAlignmentMap[i]] = sourceRead.mergeQV[queryStart + i];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->mergeQV.WriteToPos(&qvMetric[0], qvMetric.size(), offsetBegin);
}
if (metricOptions["DeletionQV"] == true) {
if (!expGroup->deletionQV.IsInitialized()) {
expGroup->deletionQV.Initialize(expGroup->experimentGroup, "DeletionQV",
true, alnArrayLength);
}
// Store DeletionQV.
std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[baseToAlignmentMap[i]] = sourceRead.deletionQV[queryStart + i];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->deletionQV.WriteToPos(&qvMetric[0], qvMetric.size(), offsetBegin);
}
if (metricOptions["DeletionTag"] == true) {
if (!expGroup->deletionTag.IsInitialized()) {
expGroup->deletionTag.Initialize(expGroup->experimentGroup, "DeletionTag",
true, alnArrayLength);
}
std::vector<char> readDeletionTagMetric;
readDeletionTagMetric.resize(readPulseMetric.size());
// Store DeletionTag.
for (i = 0; i < readDeletionTagMetric.size() - 1; i++) {
readDeletionTagMetric[i] = '-';
}
readDeletionTagMetric[i] = '\0';
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
assert(baseToAlignmentMap[i] <
static_cast<int>(readDeletionTagMetric.size()));
readDeletionTagMetric[baseToAlignmentMap[i]] =
sourceRead.deletionTag[queryStart + i];
}
readDeletionTagMetric[readDeletionTagMetric.size() - 1] = 0;
expGroup->deletionTag.WriteToPos(&readDeletionTagMetric[0],
readDeletionTagMetric.size(), offsetBegin);
}
if (metricOptions["PulseIndex"] == true) {
if (!expGroup->pulseIndex.IsInitialized()) {
expGroup->pulseIndex.Initialize(expGroup->experimentGroup, "PulseIndex",
true, alnArrayLength);
}
std::vector<uint32_t> readPulseIndexMetric;
std::fill(readPulseIndexMetric.begin(), readPulseIndexMetric.end(),
missingPulseIndex);
readPulseIndexMetric.resize(readPulseMetric.size());
// Store Pulse Index.
assert(readPulseIndexMetric.size() > 0);
for (i = 0; i < readPulseIndexMetric.size(); i++) {
readPulseIndexMetric[i] = 0;
}
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
readPulseIndexMetric[baseToAlignmentMap[i]] =
sourceRead.pulseIndex[queryStart + i];
}
readPulseIndexMetric[readPulseIndexMetric.size() - 1] = 0;
expGroup->pulseIndex.WriteToPos(&readPulseIndexMetric[0],
readPulseIndexMetric.size(), offsetBegin);
}
if (metricOptions["SubstitutionTag"] == true) {
if (!expGroup->substitutionTag.IsInitialized()) {
expGroup->substitutionTag.Initialize(
expGroup->experimentGroup, "SubstitutionTag", true, alnArrayLength);
}
std::vector<char> readSubstitutionTagMetric;
readSubstitutionTagMetric.resize(readPulseMetric.size());
// Store substitutionTag
for (i = 0; i < readSubstitutionTagMetric.size() - 1; i++) {
readSubstitutionTagMetric[i] = '-';
}
readSubstitutionTagMetric[i] = '\0';
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
readSubstitutionTagMetric[baseToAlignmentMap[i]] =
sourceRead.substitutionTag[queryStart + i];
}
readSubstitutionTagMetric[readSubstitutionTagMetric.size() - 1] = 0;
expGroup->substitutionTag.WriteToPos(&readSubstitutionTagMetric[0],
readSubstitutionTagMetric.size(),
offsetBegin);
}
if (metricOptions["SubstitutionQV"] == true) {
if (!expGroup->substitutionQV.IsInitialized()) {
expGroup->substitutionQV.Initialize(expGroup->experimentGroup,
"SubstitutionQV", true, alnArrayLength);
}
// Store start time normalized to frame rate.
std::fill(qvMetric.begin(), qvMetric.end(), missingQualityValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
qvMetric[baseToAlignmentMap[i]] = sourceRead.substitutionQV[queryStart + i];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->substitutionQV.WriteToPos(&qvMetric[0], qvMetric.size(), offsetBegin);
}
if (metricOptions["ClassifierQV"] == true) {
if (!expGroup->classifierQV.IsInitialized()) {
expGroup->classifierQV.Initialize(expGroup->experimentGroup, "ClassifierQV",
true, alnArrayLength);
}
std::fill(floatMetric.begin(), floatMetric.end(), NaN);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
floatMetric[baseToAlignmentMap[i]] =
sourceRead.classifierQV[i + queryStart];
}
qvMetric[qvMetric.size() - 1] = 0;
expGroup->classifierQV.WriteToPos(&floatMetric[0], floatMetric.size(),
offsetBegin);
}
if (metricOptions["StartFrame"] == true) {
if (!expGroup->startTime.IsInitialized()) {
expGroup->startTime.Initialize(expGroup->experimentGroup, "StartFrame",
true, alnArrayLength);
}
// StartFrame used to be computed from baseFile.preBaseFrame and
// baseFile.basWidthInFrames, whenever possible. But a more accurate
// way is to obtain StartFrame directly from pulseFile.StartFrame
// when a pulseFile is provided.
if (usePulseFile) {
assert(sourceRead.startFrame);
} else if (useBaseFile) {
if (sourceRead.startFrame) {
Free(sourceRead.startFrame);
}
sourceRead.startFrame = new unsigned int[sourceRead.length];
std::copy(sourceRead.preBaseFrames,
&sourceRead.preBaseFrames[sourceRead.length],
sourceRead.startFrame);
for (i = 0; i < sourceRead.length - 1; i++) {
sourceRead.startFrame[i + 1] += sourceRead.widthInFrames[i];
}
std::partial_sum(sourceRead.startFrame,
&sourceRead.startFrame[sourceRead.length],
sourceRead.startFrame);
}
std::fill(timeMetric.begin(), timeMetric.end(), missingPulseIndex);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
timeMetric[baseToAlignmentMap[i]] = sourceRead.startFrame[i + queryStart];
}
timeMetric[timeMetric.size() - 1] = 0;
expGroup->startTime.WriteToPos(&timeMetric[0], timeMetric.size(), offsetBegin);
}
if (metricOptions["PulseWidth"] == true) {
if (!expGroup->pulseWidth.IsInitialized()) {
expGroup->pulseWidth.Initialize(expGroup->experimentGroup, "PulseWidth",
true, alnArrayLength);
}
std::fill(frameRateMetric.begin(), frameRateMetric.end(),
missingFrameRateValue);
// For legacy reasons, it's possible the width in frames is
// stored in the bas file. If this is the case, use the width
// in frames there. Otherwise, use the width in frames stored
// in the pls file.
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[baseToAlignmentMap[i]] =
sourceRead.widthInFrames[queryStart + i];
}
frameRateMetric[frameRateMetric.size() - 1] = 0;
expGroup->pulseWidth.WriteToPos(&frameRateMetric[0], frameRateMetric.size(),
offsetBegin);
}
if (metricOptions["PreBaseFrames"] == true) {
if (!expGroup->preBaseFrames.IsInitialized()) {
expGroup->preBaseFrames.Initialize(expGroup->experimentGroup,
"PreBaseFrames", true, alnArrayLength);
}
std::fill(frameRateMetric.begin(), frameRateMetric.end(),
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[baseToAlignmentMap[i]] =
sourceRead.preBaseFrames[i + queryStart];
}
frameRateMetric[frameRateMetric.size() - 1] = 0;
expGroup->preBaseFrames.WriteToPos(&frameRateMetric[0], frameRateMetric.size(),
offsetBegin);
}
if (metricOptions["WidthInFrames"] == true) {
if (!expGroup->widthInFrames.IsInitialized()) {
expGroup->widthInFrames.Initialize(expGroup->experimentGroup,
"WidthInFrames", true, alnArrayLength);
}
// Compute width in frames.
std::fill(frameRateMetric.begin(), frameRateMetric.end(),
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[baseToAlignmentMap[i]] =
sourceRead.widthInFrames[i + queryStart];
}
frameRateMetric[frameRateMetric.size() - 1] = 0;
expGroup->widthInFrames.WriteToPos(&frameRateMetric[0], frameRateMetric.size(),
offsetBegin);
}
if (metricOptions["pkmid"] == true) {
if (!expGroup->pkmid.IsInitialized()) {
expGroup->pkmid.Initialize(expGroup->experimentGroup, "pkmid", true,
alnArrayLength);
}
for (i = 0; i < readPulseMetric.size(); i++) {
readPulseMetric[i] = NaN;
}
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
readPulseMetric[baseToAlignmentMap[i]] =
sourceRead.midSignal[i + queryStart];
}
readPulseMetric[readPulseMetric.size() - 1] = 0;
expGroup->pkmid.WriteToPos(&readPulseMetric[0], readPulseMetric.size(),
offsetBegin);
}
if (metricOptions["IPD"] == true) {
if (!expGroup->ipd.IsInitialized()) {
expGroup->ipd.Initialize(expGroup->experimentGroup, "IPD", true,
alnArrayLength);
}
std::fill(frameRateMetric.begin(), frameRateMetric.end(),
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
//
// The IPD is undefined for the first base in a read.
//
if (usePulseFile) {
if (queryStart == 0 and i == 0) {
frameRateMetric[baseToAlignmentMap[i]] = 0;
} else {
frameRateMetric[baseToAlignmentMap[i]] =
(sourceRead.startFrame[i + queryStart] -
sourceRead.startFrame[i + queryStart - 1] -
sourceRead.widthInFrames[i + queryStart - 1]);
}
} else if (useBaseFile) {
frameRateMetric[baseToAlignmentMap[i]] =
sourceRead.preBaseFrames[i + queryStart];
}
}
frameRateMetric[frameRateMetric.size() - 1] = 0;
expGroup->ipd.WriteToPos(&frameRateMetric[0], frameRateMetric.size(),
offsetBegin);
}
if (metricOptions["Light"] == true) {
if (!expGroup->light.IsInitialized()) {
expGroup->light.Initialize(expGroup->experimentGroup, "Light", true,
alnArrayLength);
}
std::fill(frameRateMetric.begin(), frameRateMetric.end(),
missingFrameRateValue);
for (i = 0; i < ungappedAlignedSequenceLength; i++) {
frameRateMetric[baseToAlignmentMap[i]] =
sourceRead.meanSignal[i + queryStart];
frameRateMetric[baseToAlignmentMap[i]] =
(frameRateMetric[baseToAlignmentMap[i]] *
sourceRead.widthInFrames[i + queryStart]);
}
frameRateMetric[frameRateMetric.size() - 1] = 0;
expGroup->light.WriteToPos(&frameRateMetric[0], frameRateMetric.size(),
offsetBegin);
}
sourceRead.Free();
Free(sourceRead.meanSignal);
Free(sourceRead.maxSignal);
Free(sourceRead.midSignal);
Free(sourceRead.startFrame);
Free(sourceRead.classifierQV);
Free(sourceRead.widthInFrames);
}
}
if (useBaseFile) {
hdfBasReader.Close();
}
if (cmpFile.readType == ReadType::CCS or useCcsOnly) {
hdfCcsReader.Close();
}
if (usePulseFile) {
hdfPlsReader.Close();
}
} // Done loading movies.
cmpReader.Close();
std::cerr << "[INFO] " << GetTimestamp() << " [" << program << "] ended." << std::endl;
}
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