/** Merge the specified seed paths.
 * @return the merged contig paths
 */
static ContigPaths mergeSeedPaths(const Lengths& lengths,
		const ContigPathMap& paths, const ContigPaths& seedPaths)
{
	if (opt::verbose > 0)
		cout << "\nMerging paths\n";

	ContigPaths out;
	out.reserve(seedPaths.size());
	for (ContigPaths::const_iterator it = seedPaths.begin();
			it != seedPaths.end(); ++it)
		out.push_back(mergePath(lengths, paths, *it));
	return out;
}
Exemple #2
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/** Find a path for the specified distance estimates.
 * @param out [out] the solution path
 */
static void handleEstimate(const Graph& g,
		const EstimateRecord& er, bool dirIdx,
		ContigPath& out)
{
	if (er.estimates[dirIdx].empty())
		return;

	ContigNode origin(er.refID, dirIdx);
	ostringstream vout_ss;
	ostream bitBucket(NULL);
	ostream& vout = opt::verbose > 0 ? vout_ss : bitBucket;
	vout << "\n* " << get(vertex_name, g, origin) << '\n';

	unsigned minNumPairs = UINT_MAX;
	// generate the reachable set
	Constraints constraints;
	for (Estimates::const_iterator iter
				= er.estimates[dirIdx].begin();
			iter != er.estimates[dirIdx].end(); ++iter) {
		ContigNode v = iter->first;
		const DistanceEst& ep = iter->second;
		minNumPairs = min(minNumPairs, ep.numPairs);
		constraints.push_back(Constraint(v,
					ep.distance + allowedError(ep.stdDev)));
	}

	vout << "Constraints:";
	printConstraints(vout, g, constraints) << '\n';

	ContigPaths solutions;
	unsigned numVisited = 0;
	constrainedSearch(g, origin, constraints, solutions, numVisited);
	bool tooComplex = numVisited >= opt::maxCost;
	bool tooManySolutions = solutions.size() > opt::maxPaths;

	set<ContigID> repeats = findRepeats(er.refID, solutions);
	if (!repeats.empty()) {
		vout << "Repeats:";
		for (set<ContigID>::const_iterator it = repeats.begin();
				it != repeats.end(); ++it)
			vout << ' ' << get(g_contigNames, *it);
		vout << '\n';
	}

	unsigned numPossiblePaths = solutions.size();
	if (numPossiblePaths > 0)
		vout << "Paths: " << numPossiblePaths << '\n';

	for (ContigPaths::iterator solIter = solutions.begin();
			solIter != solutions.end();) {
		vout << *solIter << '\n';

		// Calculate the path distance to each node and see if
		// it is within the estimated distance.
		map<ContigNode, int> distanceMap
			= makeDistanceMap(g, origin, *solIter);

		// Remove solutions whose distance estimates are not correct.
		unsigned validCount = 0, invalidCount = 0, ignoredCount = 0;
		for (Estimates::const_iterator iter
					= er.estimates[dirIdx].begin();
				iter != er.estimates[dirIdx].end(); ++iter) {
			ContigNode v = iter->first;
			const DistanceEst& ep = iter->second;
			vout << get(vertex_name, g, v) << ',' << ep << '\t';

			map<ContigNode, int>::iterator dmIter
				= distanceMap.find(v);
			if (dmIter == distanceMap.end()) {
				// This contig is a repeat.
				ignoredCount++;
				vout << "ignored\n";
				continue;
			}

			// translate distance by -overlap to match
			// coordinate space used by the estimate
			int actualDistance = dmIter->second;
			int diff = actualDistance - ep.distance;
			unsigned buffer = allowedError(ep.stdDev);
			bool invalid = (unsigned)abs(diff) > buffer;
			bool repeat = repeats.count(v.contigIndex()) > 0;
			bool ignored = invalid && repeat;
			if (ignored)
				ignoredCount++;
			else if (invalid)
				invalidCount++;
			else
				validCount++;
			vout << "dist: " << actualDistance
				<< " diff: " << diff
				<< " buffer: " << buffer
				<< " n: " << ep.numPairs
				<< (ignored ? " ignored" : invalid ? " invalid" : "")
				<< '\n';
		}

		if (invalidCount == 0 && validCount > 0)
			++solIter;
		else
			solIter = solutions.erase(solIter);
	}

	vout << "Solutions: " << solutions.size();
	if (tooComplex)
		vout << " (too complex)";
	if (tooManySolutions)
		vout << " (too many solutions)";
	vout << '\n';

	ContigPaths::iterator bestSol = solutions.end();
	int minDiff = 999999;
	for (ContigPaths::iterator solIter = solutions.begin();
			solIter != solutions.end(); ++solIter) {
		map<ContigNode, int> distanceMap
			= makeDistanceMap(g, origin, *solIter);
		int sumDiff = 0;
		for (Estimates::const_iterator iter
					= er.estimates[dirIdx].begin();
				iter != er.estimates[dirIdx].end(); ++iter) {
			ContigNode v = iter->first;
			const DistanceEst& ep = iter->second;
			if (repeats.count(v.contigIndex()) > 0)
				continue;
			map<ContigNode, int>::iterator dmIter
				= distanceMap.find(v);
			assert(dmIter != distanceMap.end());
			int actualDistance = dmIter->second;
			int diff = actualDistance - ep.distance;
			sumDiff += abs(diff);
		}

		if (sumDiff < minDiff) {
			minDiff = sumDiff;
			bestSol = solIter;
		}

		vout << *solIter
			<< " length: " << calculatePathLength(g, origin, *solIter)
			<< " sumdiff: " << sumDiff << '\n';
	}

	/** Lock the debugging stream. */
	static pthread_mutex_t coutMutex = PTHREAD_MUTEX_INITIALIZER;
	pthread_mutex_lock(&coutMutex);
	stats.totalAttempted++;
	g_minNumPairs = min(g_minNumPairs, minNumPairs);

	if (tooComplex) {
		stats.tooComplex++;
	} else if (tooManySolutions) {
		stats.tooManySolutions++;
	} else if (numPossiblePaths == 0) {
		stats.noPossiblePaths++;
	} else if (solutions.empty()) {
		stats.noValidPaths++;
	} else if (repeats.count(er.refID) > 0) {
		vout << "Repeat: " << get(vertex_name, g, origin) << '\n';
		stats.repeat++;
	} else if (solutions.size() > 1) {
		ContigPath path
			= constructAmbiguousPath(g, origin, solutions);
		if (!path.empty()) {
			if (opt::extend)
				extend(g, path.back(), back_inserter(path));
			vout << path << '\n';
			if (opt::scaffold) {
				out.insert(out.end(), path.begin(), path.end());
				g_minNumPairsUsed
					= min(g_minNumPairsUsed, minNumPairs);
			}
		}
		stats.multiEnd++;
	} else {
		assert(solutions.size() == 1);
		assert(bestSol != solutions.end());
		ContigPath& path = *bestSol;
		if (opt::verbose > 1)
			printDistanceMap(vout, g, origin, path);
		if (opt::extend)
			extend(g, path.back(), back_inserter(path));
		out.insert(out.end(), path.begin(), path.end());
		stats.uniqueEnd++;
		g_minNumPairsUsed = min(g_minNumPairsUsed, minNumPairs);
	}
	cout << vout_ss.str();
	if (!out.empty())
		assert(!out.back().ambiguous());
	pthread_mutex_unlock(&coutMutex);
}
Exemple #3
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/** Return the consensus sequence of the specified gap. */
static ContigPath fillGap(const Graph& g,
		const AmbPathConstraint& apConstraint,
		vector<bool>& seen,
		ofstream& outFasta)
{
	if (opt::verbose > 1)
		cerr << "\n* "
			<< get(vertex_name, g, apConstraint.source) << ' '
			<< apConstraint.dist << "N "
			<< get(vertex_name, g, apConstraint.dest) << '\n';

	Constraints constraints;
	constraints.push_back(Constraint(apConstraint.dest,
				apConstraint.dist + opt::distanceError));

	ContigPaths solutions;
	unsigned numVisited = 0;
	constrainedSearch(g, apConstraint.source,
			constraints, solutions, numVisited);
	bool tooComplex = numVisited >= opt::maxCost;

	for (ContigPaths::iterator solIt = solutions.begin();
			solIt != solutions.end(); solIt++)
		solIt->insert(solIt->begin(), apConstraint.source);

	ContigPath consensus;
	bool tooManySolutions = solutions.size() > opt::numBranches;
	if (tooComplex) {
		stats.tooComplex++;
		if (opt::verbose > 1)
			cerr << solutions.size() << " paths (too complex)\n";
	} else if (tooManySolutions) {
		stats.numTooManySolutions++;
		if (opt::verbose > 1)
			cerr << solutions.size() << " paths (too many)\n";
	} else if (solutions.empty()) {
		stats.numNoSolutions++;
		if (opt::verbose > 1)
			cerr << "no paths\n";
	} else if (solutions.size() == 1) {
		if (opt::verbose > 1)
			cerr << "1 path\n" << solutions.front() << '\n';
		stats.numMerged++;
	} else {
		assert(solutions.size() > 1);
		if (opt::verbose > 2)
			copy(solutions.begin(), solutions.end(),
					ostream_iterator<ContigPath>(cerr, "\n"));
		else if (opt::verbose > 1)
			cerr << solutions.size() << " paths\n";
		consensus = align(g, solutions, outFasta);
		if (!consensus.empty()) {
			stats.numMerged++;
			// Mark contigs that are used in a consensus.
			markSeen(seen, solutions, true);
			if (opt::verbose > 1)
				cerr << consensus << '\n';
		} else
			stats.notMerged++;
	}
	return consensus;
}
Exemple #4
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/* Resolve ambiguous region using pairwise alignment
 * (Needleman-Wunsch) ('solutions' contain exactly two paths, from a
 * source contig to a dest contig)
 */
static ContigPath alignPair(const Graph& g,
		const ContigPaths& solutions, ofstream& out)
{
	assert(solutions.size() == 2);
	assert(solutions[0].size() > 1);
	assert(solutions[1].size() > 1);
	assert(solutions[0].front() == solutions[1].front());
	assert(solutions[0].back() == solutions[1].back());
	ContigPath fstSol(solutions[0].begin()+1, solutions[0].end()-1);
	ContigPath sndSol(solutions[1].begin()+1, solutions[1].end()-1);

	if (fstSol.empty() || sndSol.empty()) {
		// This entire sequence may be deleted.
		const ContigPath& sol(fstSol.empty() ? sndSol : fstSol);
		assert(!sol.empty());
		Sequence consensus(mergePath(g, sol));
		assert(consensus.size() > opt::k - 1);
		string::iterator first = consensus.begin() + opt::k - 1;
		transform(first, consensus.end(), first, ::tolower);

		unsigned match = opt::k - 1;
		float identity = (float)match / consensus.size();
		if (opt::verbose > 2)
			cerr << consensus << '\n';
		if (opt::verbose > 1)
			cerr << identity
				<< (identity < opt::identity ? " (too low)\n" : "\n");
		if (identity < opt::identity)
			return ContigPath();

		unsigned coverage = calculatePathProperties(g, sol).coverage;
		ContigNode u = outputNewContig(g,
				solutions, 1, 1, consensus, coverage, out);
		ContigPath path;
		path.push_back(solutions.front().front());
		path.push_back(u);
		path.push_back(solutions.front().back());
		return path;
	}

	Sequence fstPathContig(mergePath(g, fstSol));
	Sequence sndPathContig(mergePath(g, sndSol));
	if (fstPathContig == sndPathContig) {
		// These two paths have identical sequence.
		if (fstSol.size() == sndSol.size()) {
			// A perfect match must be caused by palindrome.
			typedef ContigPath::const_iterator It;
			pair<It, It> it = mismatch(
					fstSol.begin(), fstSol.end(), sndSol.begin());
			assert(it.first != fstSol.end());
			assert(it.second != sndSol.end());
			assert(*it.first
					== get(vertex_complement, g, *it.second));
			assert(equal(it.first+1, It(fstSol.end()), it.second+1));
			if (opt::verbose > 1)
				cerr << "Palindrome: "
					<< get(vertex_contig_name, g, *it.first) << '\n';
			return solutions[0];
		} else {
			// The paths are different lengths.
			cerr << PROGRAM ": warning: "
				"Two paths have identical sequence, which may be "
				"caused by a transitive edge in the overlap graph.\n"
				<< '\t' << fstSol << '\n'
				<< '\t' << sndSol << '\n';
			return solutions[fstSol.size() > sndSol.size() ? 0 : 1];
		}
	}

	unsigned minLength = min(
			fstPathContig.length(), sndPathContig.length());
	unsigned maxLength = max(
			fstPathContig.length(), sndPathContig.length());
	float lengthRatio = (float)minLength / maxLength;
	if (lengthRatio < opt::identity) {
		if (opt::verbose > 1)
			cerr << minLength << '\t' << maxLength
				<< '\t' << lengthRatio << "\t(different length)\n";
		return ContigPath();
	}

	NWAlignment align;
	unsigned match = alignGlobal(fstPathContig, sndPathContig,
		   	align);
	float identity = (float)match / align.size();
	if (opt::verbose > 2)
		cerr << align;
	if (opt::verbose > 1)
		cerr << identity
			<< (identity < opt::identity ? " (too low)\n" : "\n");
	if (identity < opt::identity)
		return ContigPath();

	unsigned coverage = calculatePathProperties(g, fstSol).coverage
		+ calculatePathProperties(g, sndSol).coverage;
	ContigNode u = outputNewContig(g, solutions, 1, 1,
			align.consensus(), coverage, out);
	ContigPath path;
	path.push_back(solutions.front().front());
	path.push_back(u);
	path.push_back(solutions.front().back());
	return path;
}