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// ==========================================================================
// FX Tools
// ==========================================================================
// Copyright (c) 2006-2018, Knut Reinert, FU Berlin
// All rights reserved.
//
// Redistribution and use in source and binary forms, with or without
// modification, are permitted provided that the following conditions are met:
//
// * Redistributions of source code must retain the above copyright
// notice, this list of conditions and the following disclaimer.
// * Redistributions in binary form must reproduce the above copyright
// notice, this list of conditions and the following disclaimer in the
// documentation and/or other materials provided with the distribution.
// * Neither the name of Knut Reinert or the FU Berlin nor the names of
// its contributors may be used to endorse or promote products derived
// from this software without specific prior written permission.
//
// THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS "AS IS"
// AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE
// IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE
// ARE DISCLAIMED. IN NO EVENT SHALL KNUT REINERT OR THE FU BERLIN BE LIABLE
// FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL
// DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS OR
// SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER
// CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT
// LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY
// OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH
// DAMAGE.
//
// ==========================================================================
// Author: Manuel Holtgrewe <manuel.holtgrewe@fu-berlin.de>
// ==========================================================================
// Computes coverage and C+G content of a genome given a SAM or BAM file,
// window-based.
// ==========================================================================
#include <sstream>
#include <seqan/arg_parse.h>
#include <seqan/basic.h>
#include <seqan/modifier.h>
#include <seqan/sequence.h>
#include <seqan/stream.h>
#include <seqan/bam_io.h>
#include <seqan/seq_io.h>
// --------------------------------------------------------------------------
// Class FxBamCoverageOptions
// --------------------------------------------------------------------------
struct FxBamCoverageOptions
{
// Verbosity level. 0 - quiet, 1 - normal, 2 - verbose, 3 - very verbose.
int verbosity;
// Path to Genome file.
seqan::CharString inGenomePath;
// Path to SAM file.
seqan::CharString inBamPath;
// Path to output file.
seqan::CharString outPath;
// Window size to use for computation.
int32_t windowSize;
FxBamCoverageOptions() : verbosity(1), windowSize(10*1000)
{}
};
// --------------------------------------------------------------------------
// Class BinData
// --------------------------------------------------------------------------
struct BinData
{
// Number of reads whose alignment starts here.
unsigned coverage;
// Length of underlying sequence.
unsigned length;
// Percentag C+G.
double cgContent;
BinData() : coverage(0), length(0), cgContent(0)
{}
};
// --------------------------------------------------------------------------
// Function parseArgs()
// --------------------------------------------------------------------------
seqan::ArgumentParser::ParseResult
parseArgs(FxBamCoverageOptions & options,
int argc,
char const ** argv)
{
seqan::ArgumentParser parser("fx_bam_coverage");
setShortDescription(parser, "Read Coverage Computation.");
setCategory(parser, "Utilities");
setVersion(parser, SEQAN_APP_VERSION " [" SEQAN_REVISION "]");
setDate(parser, SEQAN_DATE);
addUsageLine(parser,
"[\\fIOPTIONS\\fP] \\fB-o\\fP \\fIOUT.coverage.tsv\\fP \\fB-r\\fP \\fIGENOME.fa\\fP "
"\\fB-m\\fP \\fIMAPPING.bam\\fP");
addDescription(parser, "Compute read coverage and C+G content for a genome.");
// Two input files: Genome, and mapping.
addOption(parser, seqan::ArgParseOption("r", "in-reference", "Path to the reference file.",
seqan::ArgParseArgument::INPUT_FILE, "IN.fa"));
setValidValues(parser, "in-reference", "fasta fa");
setRequired(parser, "in-reference");
addOption(parser, seqan::ArgParseOption("m", "in-mapping", "Path to the mapping file to analyze.",
seqan::ArgParseArgument::INPUT_FILE));
setValidValues(parser, "in-mapping", "sam bam");
setRequired(parser, "in-mapping");
// TODO(holtgrew): I want a custom help text!
// addOption(parser, seqan::ArgParseOption("h", "help", "This helpful screen."));
addOption(parser, seqan::ArgParseOption("v", "verbose", "Verbose, log to STDERR."));
hideOption(parser, "verbose");
addOption(parser, seqan::ArgParseOption("vv", "very-verbose", "Very verbose, log to STDERR."));
hideOption(parser, "very-verbose");
addSection(parser, "Main Options");
addOption(parser, seqan::ArgParseOption("w", "window-size", "Set the size of the non-overlapping windows in base pairs.", seqan::ArgParseArgument::INTEGER, "NUM"));
setDefaultValue(parser, "window-size", options.windowSize);
addSection(parser, "Output Options");
addOption(parser, seqan::ArgParseOption("o", "out-path", "Path to the resulting file. If omitted, result is printed to stdout.", seqan::ArgParseArgument::OUTPUT_FILE, "TSV"));
setRequired(parser, "out-path");
setValidValues(parser, "out-path", ".coverage.tsv");
seqan::ArgumentParser::ParseResult res = parse(parser, argc, argv);
if (res == seqan::ArgumentParser::PARSE_OK)
{
getOptionValue(options.inGenomePath, parser, "in-reference");
getOptionValue(options.inBamPath, parser, "in-mapping");
getOptionValue(options.outPath, parser, "out-path");
getOptionValue(options.windowSize, parser, "window-size");
if (isSet(parser, "verbose"))
options.verbosity = 2;
if (isSet(parser, "very-verbose"))
options.verbosity = 3;
}
return res;
}
// ---------------------------------------------------------------------------
// Function main()
// ---------------------------------------------------------------------------
int main(int argc, char const ** argv)
{
double startTime = seqan::sysTime();
// -----------------------------------------------------------------------
// Parse command line.
// -----------------------------------------------------------------------
FxBamCoverageOptions options;
seqan::ArgumentParser::ParseResult res = parseArgs(options, argc, argv);
if (res != seqan::ArgumentParser::PARSE_OK)
return res == seqan::ArgumentParser::PARSE_ERROR; // 1 on errors, 0 otherwise
// -----------------------------------------------------------------------
// Show options.
// -----------------------------------------------------------------------
if (options.verbosity >= 1)
{
std::cerr << "____OPTIONS___________________________________________________________________\n"
<< "\n"
<< "VERBOSITY " << options.verbosity << "\n"
<< "GENOME " << options.inGenomePath << "\n"
<< "SAM/BAM " << options.inBamPath << "\n"
<< "OUT " << options.outPath << "\n"
<< "WINDOW SIZE " << options.windowSize << "\n";
}
// -----------------------------------------------------------------------
// Load Genome FAI Index
// -----------------------------------------------------------------------
std::cerr << "\n"
<< "___PREPARATION____________________________________________________________________\n"
<< "\n";
seqan::FaiIndex faiIndex;
if (!open(faiIndex, toCString(options.inGenomePath)))
{
std::cerr << "Indexing GENOME file " << options.inGenomePath << " ...";
if (!build(faiIndex, toCString(options.inGenomePath)))
{
std::cerr << "Could not build FAI index of " << options.inGenomePath << "!\n";
return 1;
}
save(faiIndex);
}
std::cerr << " OK\n";
// Prepare bins.
seqan::String<seqan::String<BinData> > bins;
resize(bins, numSeqs(faiIndex));
// -----------------------------------------------------------------------
// Compute C+G content
// -----------------------------------------------------------------------
std::cerr << "\n"
<< "___C+G CONTENT COMPUTATION________________________________________________________\n"
<< "\n";
seqan::Dna5String contigSeq;
for (unsigned i = 0; i < numSeqs(faiIndex); ++i)
{
std::cerr << "[" << sequenceName(faiIndex, i) << "] ...";
unsigned numBins = (sequenceLength(faiIndex, i) + options.windowSize - 1) / options.windowSize;
resize(bins[i], numBins);
clear(contigSeq);
readSequence(contigSeq, faiIndex, i);
for (unsigned bin = 0; bin < numBins; ++bin)
{
unsigned cgCounter = 0;
unsigned binSize = 0;
bins[i][bin].length = options.windowSize;
if ((bin + 1) * options.windowSize > length(contigSeq))
bins[i][bin].length = length(contigSeq) - bin * options.windowSize;
for (unsigned pos = bin * options.windowSize; pos < length(contigSeq) && pos < (bin + 1) * options.windowSize; ++pos, ++binSize)
cgCounter += (contigSeq[pos] == 'C' || contigSeq[pos] == 'G');
if (binSize == 0u) // prevent div-by-zero below
binSize = 1;
bins[i][bin].cgContent = 1.0 * cgCounter / binSize;
}
std::cerr << "DONE\n";
}
// -----------------------------------------------------------------------
// Compute Coverage
// -----------------------------------------------------------------------
std::cerr << "\n"
<< "___COVERAGE COMPUTATION_________________________________________________________\n"
<< "\n"
<< "Computing Coverage...";
seqan::BamFileIn bamFile;
if (!open(bamFile, toCString(options.inBamPath)))
{
std::cerr << "Could not open " << options.inBamPath << "!\n";
return 1;
}
seqan::BamHeader header;
readHeader(header, bamFile);
seqan::BamAlignmentRecord record;
while (!atEnd(bamFile))
{
readRecord(record, bamFile);
if (hasFlagUnmapped(record) || hasFlagSecondary(record) || record.rID == seqan::BamAlignmentRecord::INVALID_REFID)
continue; // Skip these records.
int contigId = 0;
seqan::CharString const & contigName = contigNames(context(bamFile))[record.rID];
if (!getIdByName(contigId, faiIndex, contigName))
{
std::cerr << "ERROR: Alignment to unknown contig " << contigName << "!\n";
return 1;
}
unsigned binNo = record.beginPos / options.windowSize;
bins[contigId][binNo].coverage += 1;
}
std::cerr << "DONE\n";
// -----------------------------------------------------------------------
// Write Output
// -----------------------------------------------------------------------
std::ostream * out = &std::cout;
std::ofstream outFile;
if (options.outPath != "-")
{
outFile.open(toCString(options.outPath), std::ios::binary | std::ios::out);
if (!outFile.good())
{
std::cerr << "ERROR: Could not open output file " << options.outPath << "!\n";
return 1;
}
out = &outFile;
}
(*out) << "#BIN\tREF_NAME\tREF_BIN\tBIN_BEGIN\tBIN_LENGTH\tCOVERAGE\tCG_CONTENT\n";
for (unsigned i = 0, globalBin = 0; i < length(bins); ++i)
{
for (unsigned refBin = 0; refBin < length(bins[i]); ++refBin, ++globalBin)
{
(*out) << globalBin << '\t'
<< sequenceName(faiIndex, i) << '\t'
<< refBin << '\t'
<< refBin * options.windowSize << '\t'
<< bins[i][refBin].length << '\t'
<< bins[i][refBin].coverage << '\t'
<< bins[i][refBin].cgContent << '\n';
}
}
if (options.verbosity >= 2)
std::cerr << "Took " << (seqan::sysTime() - startTime) << " s\n";
return 0;
}
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