Esempio n. 1
0
static void testPinchIteratorFromList(CuTest *testCase) {
    for (int64_t test = 0; test < 100; test++) {
        stList *pairwiseAlignments = getRandomPairwiseAlignments();
        st_logInfo("Doing a random pinch iterator from list test %" PRIi64 " with %" PRIi64 " alignments\n", test, stList_length(pairwiseAlignments));
        //Get an iterator
        stPinchIterator *pinchIterator = stPinchIterator_constructFromList(pairwiseAlignments);
        //Now test it
        testIterator(testCase, pinchIterator, pairwiseAlignments);
        //Cleanup
        stPinchIterator_destruct(pinchIterator);
        stList_destruct(pairwiseAlignments);
    }
}
Esempio n. 2
0
int main(int argc, char *argv[]) {
    /*
     * Script for adding alignments to cactus tree.
     */
    int64_t startTime;
    stKVDatabaseConf *kvDatabaseConf;
    CactusDisk *cactusDisk;
    int key, k;

    bool (*filterFn)(stPinchSegment *, stPinchSegment *) = NULL;
    stSet *outgroupThreads = NULL;

    /*
     * Arguments/options
     */
    char * logLevelString = NULL;
    char * alignmentsFile = NULL;
    char * constraintsFile = NULL;
    char * cactusDiskDatabaseString = NULL;
    char * lastzArguments = "";
    int64_t minimumSequenceLengthForBlast = 1;

    //Parameters for annealing/melting rounds
    int64_t *annealingRounds = NULL;
    int64_t annealingRoundsLength = 0;
    int64_t *meltingRounds = NULL;
    int64_t meltingRoundsLength = 0;

    //Parameters for melting
    float maximumAdjacencyComponentSizeRatio = 10;
    int64_t blockTrim = 0;
    int64_t alignmentTrimLength = 0;
    int64_t *alignmentTrims = NULL;
    int64_t chainLengthForBigFlower = 1000000;
    int64_t longChain = 2;
    int64_t minLengthForChromosome = 1000000;
    float proportionOfUnalignedBasesForNewChromosome = 0.8;
    bool breakChainsAtReverseTandems = 1;
    int64_t maximumMedianSequenceLengthBetweenLinkedEnds = INT64_MAX;
    bool realign = 0;
    char *realignArguments = "";
    bool removeRecoverableChains = false;
    bool (*recoverableChainsFilter)(stCactusEdgeEnd *, Flower *) = NULL;
    int64_t maxRecoverableChainsIterations = 1;
    int64_t maxRecoverableChainLength = INT64_MAX;

    //Parameters for removing ancient homologies
    bool doPhylogeny = false;
    int64_t phylogenyNumTrees = 1;
    enum stCaf_RootingMethod phylogenyRootingMethod = BEST_RECON;
    enum stCaf_ScoringMethod phylogenyScoringMethod = COMBINED_LIKELIHOOD;
    double breakpointScalingFactor = 1.0;
    bool phylogenySkipSingleCopyBlocks = 0;
    int64_t phylogenyMaxBaseDistance = 1000;
    int64_t phylogenyMaxBlockDistance = 100;
    bool phylogenyKeepSingleDegreeBlocks = 0;
    stList *phylogenyTreeBuildingMethods = stList_construct();
    enum stCaf_TreeBuildingMethod defaultMethod = GUIDED_NEIGHBOR_JOINING;
    stList_append(phylogenyTreeBuildingMethods, &defaultMethod);
    double phylogenyCostPerDupPerBase = 0.2;
    double phylogenyCostPerLossPerBase = 0.2;
    const char *debugFileName = NULL;
    const char *referenceEventHeader = NULL;
    double phylogenyDoSplitsWithSupportHigherThanThisAllAtOnce = 1.0;
    int64_t numTreeBuildingThreads = 2;
    int64_t minimumBlockDegreeToCheckSupport = 10;
    double minimumBlockHomologySupport = 0.7;
    double nucleotideScalingFactor = 1.0;
    HomologyUnitType phylogenyHomologyUnitType = BLOCK;
    enum stCaf_DistanceCorrectionMethod phylogenyDistanceCorrectionMethod = JUKES_CANTOR;
    bool sortAlignments = false;

    ///////////////////////////////////////////////////////////////////////////
    // (0) Parse the inputs handed by genomeCactus.py / setup stuff.
    ///////////////////////////////////////////////////////////////////////////

    while (1) {
        static struct option long_options[] = { { "logLevel", required_argument, 0, 'a' }, { "alignments", required_argument, 0, 'b' }, {
                "cactusDisk", required_argument, 0, 'c' }, { "lastzArguments", required_argument, 0, 'd' },
                { "help", no_argument, 0, 'h' }, { "annealingRounds", required_argument, 0, 'i' }, { "trim", required_argument, 0, 'k' }, {
                        "trimChange", required_argument, 0, 'l', }, { "minimumTreeCoverage", required_argument, 0, 'm' }, { "blockTrim",
                        required_argument, 0, 'n' }, { "deannealingRounds", required_argument, 0, 'o' }, { "minimumDegree",
                        required_argument, 0, 'p' }, { "minimumIngroupDegree", required_argument, 0, 'q' }, {
                        "minimumOutgroupDegree", required_argument, 0, 'r' }, { "alignmentFilter", required_argument, 0, 't' }, {
                        "minimumSequenceLengthForBlast", required_argument, 0, 'v' }, { "maxAdjacencyComponentSizeRatio",
                        required_argument, 0, 'w' }, { "constraints", required_argument, 0, 'x' }, { "minLengthForChromosome",
                        required_argument, 0, 'y' }, { "proportionOfUnalignedBasesForNewChromosome", required_argument, 0, 'z' },
                        { "maximumMedianSequenceLengthBetweenLinkedEnds", required_argument, 0, 'A' },
                        { "realign", no_argument, 0, 'B' }, { "realignArguments", required_argument, 0, 'C' },
                        { "phylogenyNumTrees", required_argument, 0, 'D' },
                        { "phylogenyRootingMethod", required_argument, 0, 'E' },
                        { "phylogenyScoringMethod", required_argument, 0, 'F' },
                        { "phylogenyBreakpointScalingFactor", required_argument, 0, 'G' },
                        { "phylogenySkipSingleCopyBlocks", no_argument, 0, 'H' },
                        { "phylogenyMaxBaseDistance", required_argument, 0, 'I' },
                        { "phylogenyMaxBlockDistance", required_argument, 0, 'J' },
                        { "phylogenyDebugFile", required_argument, 0, 'K' },
                        { "phylogenyKeepSingleDegreeBlocks", no_argument, 0, 'L' },
                        { "phylogenyTreeBuildingMethod", required_argument, 0, 'M' },
                        { "phylogenyCostPerDupPerBase", required_argument, 0, 'N' },
                        { "phylogenyCostPerLossPerBase", required_argument, 0, 'O' },
                        { "referenceEventHeader", required_argument, 0, 'P' },
                        { "phylogenyDoSplitsWithSupportHigherThanThisAllAtOnce", required_argument, 0, 'Q' },
                        { "numTreeBuildingThreads", required_argument, 0, 'R' },
                        { "phylogeny", no_argument, 0, 'S' },
                        { "minimumBlockHomologySupport", required_argument, 0, 'T' },
                        { "phylogenyNucleotideScalingFactor", required_argument, 0, 'U' },
                        { "minimumBlockDegreeToCheckSupport", required_argument, 0, 'V' },
                        { "removeRecoverableChains", required_argument, 0, 'W' },
                        { "minimumNumberOfSpecies", required_argument, 0, 'X' },
                        { "phylogenyHomologyUnitType", required_argument, 0, 'Y' },
                        { "phylogenyDistanceCorrectionMethod", required_argument, 0, 'Z' },
                        { "maxRecoverableChainsIterations", required_argument, 0, '1' },
                        { "maxRecoverableChainLength", required_argument, 0, '2' },
                        { 0, 0, 0, 0 } };

        int option_index = 0;

        key = getopt_long(argc, argv, "a:b:c:hi:k:m:n:o:p:q:r:stv:w:x:y:z:A:BC:D:E:", long_options, &option_index);

        if (key == -1) {
            break;
        }

        switch (key) {
            case 'a':
                logLevelString = stString_copy(optarg);
                st_setLogLevelFromString(logLevelString);
                break;
            case 'b':
                alignmentsFile = stString_copy(optarg);
                break;
            case 'c':
                cactusDiskDatabaseString = stString_copy(optarg);
                break;
            case 'd':
                lastzArguments = stString_copy(optarg);
                break;
            case 'h':
                usage();
                return 0;
            case 'i':
                annealingRounds = getInts(optarg, &annealingRoundsLength);
                break;
            case 'o':
                meltingRounds = getInts(optarg, &meltingRoundsLength);
                break;
            case 'k':
                alignmentTrims = getInts(optarg, &alignmentTrimLength);
                break;
            case 'm':
                k = sscanf(optarg, "%f", &minimumTreeCoverage);
                assert(k == 1);
                break;
            case 'n':
                k = sscanf(optarg, "%" PRIi64 "", &blockTrim);
                assert(k == 1);
                break;
            case 'p':
                k = sscanf(optarg, "%" PRIi64 "", &minimumDegree);
                assert(k == 1);
                break;
            case 'q':
                k = sscanf(optarg, "%" PRIi64 "", &minimumIngroupDegree);
                assert(k == 1);
                break;
            case 'r':
                k = sscanf(optarg, "%" PRIi64 "", &minimumOutgroupDegree);
                assert(k == 1);
                break;
            case 't':
                if (strcmp(optarg, "singleCopyOutgroup") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_filterByOutgroup;
                } else if (strcmp(optarg, "relaxedSingleCopyOutgroup") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_relaxedFilterByOutgroup;
                } else if (strcmp(optarg, "singleCopy") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_filterByRepeatSpecies;
                } else if (strcmp(optarg, "relaxedSingleCopy") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_relaxedFilterByRepeatSpecies;
                } else if (strcmp(optarg, "singleCopyChr") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_singleCopyChr;
                } else if (strcmp(optarg, "singleCopyIngroup") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_singleCopyIngroup;
                } else if (strcmp(optarg, "relaxedSingleCopyIngroup") == 0) {
                    sortAlignments = true;
                    filterFn = stCaf_relaxedSingleCopyIngroup;
                } else if (strcmp(optarg, "none") == 0) {
                    sortAlignments = false;
                    filterFn = NULL;
                } else {
                    st_errAbort("Could not recognize alignmentFilter option %s", optarg);
                }
                break;
            case 'v':
                k = sscanf(optarg, "%" PRIi64 "", &minimumSequenceLengthForBlast);
                assert(k == 1);
                break;
            case 'w':
                k = sscanf(optarg, "%f", &maximumAdjacencyComponentSizeRatio);
                assert(k == 1);
                break;
            case 'x':
                constraintsFile = stString_copy(optarg);
                break;
            case 'y':
                k = sscanf(optarg, "%" PRIi64 "", &minLengthForChromosome);
                assert(k == 1);
                break;
            case 'z':
                k = sscanf(optarg, "%f", &proportionOfUnalignedBasesForNewChromosome);
                assert(k == 1);
                break;
            case 'A':
                k = sscanf(optarg, "%" PRIi64 "", &maximumMedianSequenceLengthBetweenLinkedEnds);
                assert(k == 1);
                break;
            case 'B':
                realign = 1;
                break;
            case 'C':
                realignArguments = stString_copy(optarg);
                break;
            case 'D':
                k = sscanf(optarg, "%" PRIi64, &phylogenyNumTrees);
                assert(k == 1);
                break;
            case 'E':
                if (!strcmp(optarg, "outgroupBranch")) {
                    phylogenyRootingMethod = OUTGROUP_BRANCH;
                } else if (!strcmp(optarg, "longestBranch")) {
                    phylogenyRootingMethod = LONGEST_BRANCH;
                } else if (!strcmp(optarg, "bestRecon")) {
                    phylogenyRootingMethod = BEST_RECON;
                } else {
                    st_errAbort("Invalid tree rooting method: %s", optarg);
                }
                break;
            case 'F':
                if (!strcmp(optarg, "reconCost")) {
                    phylogenyScoringMethod = RECON_COST;
                } else if (!strcmp(optarg, "nucLikelihood")) {
                    phylogenyScoringMethod = NUCLEOTIDE_LIKELIHOOD;
                } else if (!strcmp(optarg, "reconLikelihood")) {
                    phylogenyScoringMethod = RECON_LIKELIHOOD;
                } else if (!strcmp(optarg, "combinedLikelihood")) {
                    phylogenyScoringMethod = COMBINED_LIKELIHOOD;
                } else {
                    st_errAbort("Invalid tree scoring method: %s", optarg);
                }
                break;
            case 'G':
                k = sscanf(optarg, "%lf", &breakpointScalingFactor);
                assert(k == 1);
                break;
            case 'H':
                phylogenySkipSingleCopyBlocks = true;
                break;
            case 'I':
                k = sscanf(optarg, "%" PRIi64, &phylogenyMaxBaseDistance);
                assert(k == 1);
                break;
            case 'J':
                k = sscanf(optarg, "%" PRIi64, &phylogenyMaxBlockDistance);
                assert(k == 1);
                break;
            case 'K':
                debugFileName = stString_copy(optarg);
                break;
            case 'L':
                phylogenyKeepSingleDegreeBlocks = true;
                break;
            case 'M':
                // clear the default setting of the list
                stList_destruct(phylogenyTreeBuildingMethods);
                phylogenyTreeBuildingMethods = stList_construct();
                stList *methodStrings = stString_splitByString(optarg, ",");

                for (int64_t i = 0; i < stList_length(methodStrings); i++) {
                    char *methodString = stList_get(methodStrings, i);
                    enum stCaf_TreeBuildingMethod *method = st_malloc(sizeof(enum stCaf_TreeBuildingMethod));
                    if (strcmp(methodString, "neighborJoining") == 0) {
                        *method = NEIGHBOR_JOINING;
                    } else if (strcmp(methodString, "guidedNeighborJoining") == 0) {
                        *method = GUIDED_NEIGHBOR_JOINING;
                    } else if (strcmp(methodString, "splitDecomposition") == 0) {
                        *method = SPLIT_DECOMPOSITION;
                    } else if (strcmp(methodString, "strictSplitDecomposition") == 0) {
                        *method = STRICT_SPLIT_DECOMPOSITION;
                    } else if (strcmp(methodString, "removeBadChains") == 0) {
                        *method = REMOVE_BAD_CHAINS;
                    } else {
                        st_errAbort("Unknown tree building method: %s", methodString);
                    }
                    stList_append(phylogenyTreeBuildingMethods, method);
                }
                stList_destruct(methodStrings);
                break;
            case 'N':
                k = sscanf(optarg, "%lf", &phylogenyCostPerDupPerBase);
                assert(k == 1);
                break;
            case 'O':
                k = sscanf(optarg, "%lf", &phylogenyCostPerLossPerBase);
                assert(k == 1);
                break;
            case 'P':
                referenceEventHeader = stString_copy(optarg);
                break;
            case 'Q':
                k = sscanf(optarg, "%lf", &phylogenyDoSplitsWithSupportHigherThanThisAllAtOnce);
                assert(k == 1);
                break;
            case 'R':
                k = sscanf(optarg, "%" PRIi64, &numTreeBuildingThreads);
                assert(k == 1);
                break;
            case 'S':
                doPhylogeny = true;
                break;
            case 'T':
                k = sscanf(optarg, "%lf", &minimumBlockHomologySupport);
                assert(k == 1);
                assert(minimumBlockHomologySupport <= 1.0);
                assert(minimumBlockHomologySupport >= 0.0);
                break;
            case 'U':
                k = sscanf(optarg, "%lf", &nucleotideScalingFactor);
                assert(k == 1);
                break;
            case 'V':
                k = sscanf(optarg, "%" PRIi64, &minimumBlockDegreeToCheckSupport);
                assert(k == 1);
                break;
            case 'W':
                if (strcmp(optarg, "1") == 0) {
                    removeRecoverableChains = true;
                    recoverableChainsFilter = NULL;
                } else if (strcmp(optarg, "unequalNumberOfIngroupCopies") == 0) {
                    removeRecoverableChains = true;
                    recoverableChainsFilter = stCaf_chainHasUnequalNumberOfIngroupCopies;
                } else if (strcmp(optarg, "unequalNumberOfIngroupCopiesOrNoOutgroup") == 0) {
                    removeRecoverableChains = true;
                    recoverableChainsFilter = stCaf_chainHasUnequalNumberOfIngroupCopiesOrNoOutgroup;
                } else if (strcmp(optarg, "0") == 0) {
                    removeRecoverableChains = false;
                } else {
                    st_errAbort("Could not parse removeRecoverableChains argument");
                }
                break;
            case 'X':
                k = sscanf(optarg, "%" PRIi64, &minimumNumberOfSpecies);
                if (k != 1) {
                    st_errAbort("Error parsing the minimumNumberOfSpecies argument");
                }
                break;
            case 'Y':
                if (strcmp(optarg, "chain") == 0) {
                    phylogenyHomologyUnitType = CHAIN;
                } else if (strcmp(optarg, "block") == 0) {
                    phylogenyHomologyUnitType = BLOCK;
                } else {
                    st_errAbort("Could not parse the phylogenyHomologyUnitType argument");
                }
                break;
            case 'Z':
                if (strcmp(optarg, "jukesCantor") == 0) {
                    phylogenyDistanceCorrectionMethod = JUKES_CANTOR;
                } else if (strcmp(optarg, "none") == 0 ) {
                    phylogenyDistanceCorrectionMethod = NONE;
                } else {
                    st_errAbort("Could not parse the phylogenyDistanceCorrectionMethod argument");
                }
                break;
            case '1':
                k = sscanf(optarg, "%" PRIi64, &maxRecoverableChainsIterations);
                if (k != 1) {
                    st_errAbort("Error parsing the maxRecoverableChainsIterations argument");
                }
                break;
            case '2':
                k = sscanf(optarg, "%" PRIi64, &maxRecoverableChainLength);
                if (k != 1) {
                    st_errAbort("Error parsing the maxRecoverableChainLength argument");
                }
                break;
            default:
                usage();
                return 1;
        }
    }

    ///////////////////////////////////////////////////////////////////////////
    // (0) Check the inputs.
    ///////////////////////////////////////////////////////////////////////////

    assert(cactusDiskDatabaseString != NULL);
    assert(minimumTreeCoverage >= 0.0);
    assert(minimumTreeCoverage <= 1.0);
    assert(blockTrim >= 0);
    assert(annealingRoundsLength >= 0);
    for (int64_t i = 0; i < annealingRoundsLength; i++) {
        assert(annealingRounds[i] >= 0);
    }
    assert(meltingRoundsLength >= 0);
    for (int64_t i = 1; i < meltingRoundsLength; i++) {
        assert(meltingRounds[i - 1] < meltingRounds[i]);
        assert(meltingRounds[i - 1] >= 1);
    }
    assert(alignmentTrimLength >= 0);
    for (int64_t i = 0; i < alignmentTrimLength; i++) {
        assert(alignmentTrims[i] >= 0);
    }
    assert(minimumOutgroupDegree >= 0);
    assert(minimumIngroupDegree >= 0);

    //////////////////////////////////////////////
    //Set up logging
    //////////////////////////////////////////////

    st_setLogLevelFromString(logLevelString);

    //////////////////////////////////////////////
    //Log (some of) the inputs
    //////////////////////////////////////////////

    st_logInfo("Flower disk name : %s\n", cactusDiskDatabaseString);

    //////////////////////////////////////////////
    //Load the database
    //////////////////////////////////////////////

    kvDatabaseConf = stKVDatabaseConf_constructFromString(cactusDiskDatabaseString);
    cactusDisk = cactusDisk_construct(kvDatabaseConf, 0);
    st_logInfo("Set up the flower disk\n");

    ///////////////////////////////////////////////////////////////////////////
    // Sort the constraints
    ///////////////////////////////////////////////////////////////////////////

    stPinchIterator *pinchIteratorForConstraints = NULL;
    if (constraintsFile != NULL) {
        pinchIteratorForConstraints = stPinchIterator_constructFromFile(constraintsFile);
        st_logInfo("Created an iterator for the alignment constaints from file: %s\n", constraintsFile);
    }

    ///////////////////////////////////////////////////////////////////////////
    // Do the alignment
    ///////////////////////////////////////////////////////////////////////////

    startTime = time(NULL);

    stList *flowers = flowerWriter_parseFlowersFromStdin(cactusDisk);
    if (alignmentsFile == NULL) {
        cactusDisk_preCacheStrings(cactusDisk, flowers);
    }
    char *tempFile1 = NULL;
    for (int64_t i = 0; i < stList_length(flowers); i++) {
        flower = stList_get(flowers, i);
        if (!flower_builtBlocks(flower)) { // Do nothing if the flower already has defined blocks
            st_logDebug("Processing flower: %lli\n", flower_getName(flower));

            stCaf_setFlowerForAlignmentFiltering(flower);

            //Set up the graph and add the initial alignments
            stPinchThreadSet *threadSet = stCaf_setup(flower);

            //Build the set of outgroup threads
            outgroupThreads = stCaf_getOutgroupThreads(flower, threadSet);

            //Setup the alignments
            stPinchIterator *pinchIterator;
            stList *alignmentsList = NULL;
            if (alignmentsFile != NULL) {
                assert(i == 0);
                assert(stList_length(flowers) == 1);
                if (sortAlignments) {
                    tempFile1 = getTempFile();
                    stCaf_sortCigarsFileByScoreInDescendingOrder(alignmentsFile, tempFile1);
                    pinchIterator = stPinchIterator_constructFromFile(tempFile1);
                } else {
                    pinchIterator = stPinchIterator_constructFromFile(alignmentsFile);
                }
            } else {
                if (tempFile1 == NULL) {
                    tempFile1 = getTempFile();
                }
                alignmentsList = stCaf_selfAlignFlower(flower, minimumSequenceLengthForBlast, lastzArguments, realign, realignArguments, tempFile1);
                if (sortAlignments) {
                    stCaf_sortCigarsByScoreInDescendingOrder(alignmentsList);
                }
                st_logDebug("Ran lastz and have %" PRIi64 " alignments\n", stList_length(alignmentsList));
                pinchIterator = stPinchIterator_constructFromList(alignmentsList);
            }

            for (int64_t annealingRound = 0; annealingRound < annealingRoundsLength; annealingRound++) {
                int64_t minimumChainLength = annealingRounds[annealingRound];
                int64_t alignmentTrim = annealingRound < alignmentTrimLength ? alignmentTrims[annealingRound] : 0;
                st_logDebug("Starting annealing round with a minimum chain length of %" PRIi64 " and an alignment trim of %" PRIi64 "\n", minimumChainLength, alignmentTrim);
                stPinchIterator_setTrim(pinchIterator, alignmentTrim);

                //Add back in the constraints
                if (pinchIteratorForConstraints != NULL) {
                    stCaf_anneal(threadSet, pinchIteratorForConstraints, filterFn);
                }

                //Do the annealing
                if (annealingRound == 0) {
                    stCaf_anneal(threadSet, pinchIterator, filterFn);
                } else {
                    stCaf_annealBetweenAdjacencyComponents(threadSet, pinchIterator, filterFn);
                }

                // Dump the block degree and length distribution to a file
                if (debugFileName != NULL) {
                    dumpBlockInfo(threadSet, stString_print("%s-blockStats-preMelting", debugFileName));
                }

                printf("Sequence graph statistics after annealing:\n");
                printThreadSetStatistics(threadSet, flower, stdout);

                // Check for poorly-supported blocks--those that have
                // been transitively aligned together but with very
                // few homologies supporting the transitive
                // alignment. These "megablocks" can snarl up the
                // graph so that a lot of extra gets thrown away in
                // the first melting step.
                stPinchThreadSetBlockIt blockIt = stPinchThreadSet_getBlockIt(threadSet);
                stPinchBlock *block;
                while ((block = stPinchThreadSetBlockIt_getNext(&blockIt)) != NULL) {
                    if (stPinchBlock_getDegree(block) > minimumBlockDegreeToCheckSupport) {
                        uint64_t supportingHomologies = stPinchBlock_getNumSupportingHomologies(block);
                        uint64_t possibleSupportingHomologies = numPossibleSupportingHomologies(block, flower);
                        double support = ((double) supportingHomologies) / possibleSupportingHomologies;
                        if (support < minimumBlockHomologySupport) {
                            fprintf(stdout, "Destroyed a megablock with degree %" PRIi64
                                    " and %" PRIi64 " supporting homologies out of a maximum "
                                    "of %" PRIi64 " (%lf%%).\n", stPinchBlock_getDegree(block),
                                    supportingHomologies, possibleSupportingHomologies, support);
                            stPinchBlock_destruct(block);
                        }
                    }
                }

                //Do the melting rounds
                for (int64_t meltingRound = 0; meltingRound < meltingRoundsLength; meltingRound++) {
                    int64_t minimumChainLengthForMeltingRound = meltingRounds[meltingRound];
                    st_logDebug("Starting melting round with a minimum chain length of %" PRIi64 " \n", minimumChainLengthForMeltingRound);
                    if (minimumChainLengthForMeltingRound >= minimumChainLength) {
                        break;
                    }
                    stCaf_melt(flower, threadSet, NULL, 0, minimumChainLengthForMeltingRound, 0, INT64_MAX);
                } st_logDebug("Last melting round of cycle with a minimum chain length of %" PRIi64 " \n", minimumChainLength);
                stCaf_melt(flower, threadSet, NULL, 0, minimumChainLength, breakChainsAtReverseTandems, maximumMedianSequenceLengthBetweenLinkedEnds);
                //This does the filtering of blocks that do not have the required species/tree-coverage/degree.
                stCaf_melt(flower, threadSet, blockFilterFn, blockTrim, 0, 0, INT64_MAX);
            }

            if (removeRecoverableChains) {
                stCaf_meltRecoverableChains(flower, threadSet, breakChainsAtReverseTandems, maximumMedianSequenceLengthBetweenLinkedEnds, recoverableChainsFilter, maxRecoverableChainsIterations, maxRecoverableChainLength);
            }
            if (debugFileName != NULL) {
                dumpBlockInfo(threadSet, stString_print("%s-blockStats-postMelting", debugFileName));
            }

            printf("Sequence graph statistics after melting:\n");
            printThreadSetStatistics(threadSet, flower, stdout);

            // Build a tree for each block, then use each tree to
            // partition the homologies between the ingroups sequences
            // into those that occur before the speciation with the
            // outgroup and those which occur late.

            if (stSet_size(outgroupThreads) > 0 && doPhylogeny) {
                st_logDebug("Starting to build trees and partition ingroup homologies\n");
                stHash *threadStrings = stCaf_getThreadStrings(flower, threadSet);
                st_logDebug("Got sets of thread strings and set of threads that are outgroups\n");
                stCaf_PhylogenyParameters params;
                params.distanceCorrectionMethod = phylogenyDistanceCorrectionMethod;
                params.treeBuildingMethods = phylogenyTreeBuildingMethods;
                params.rootingMethod = phylogenyRootingMethod;
                params.scoringMethod = phylogenyScoringMethod;
                params.breakpointScalingFactor = breakpointScalingFactor;
                params.nucleotideScalingFactor = nucleotideScalingFactor;
                params.skipSingleCopyBlocks = phylogenySkipSingleCopyBlocks;
                params.keepSingleDegreeBlocks = phylogenyKeepSingleDegreeBlocks;
                params.costPerDupPerBase = phylogenyCostPerDupPerBase;
                params.costPerLossPerBase = phylogenyCostPerLossPerBase;
                params.maxBaseDistance = phylogenyMaxBaseDistance;
                params.maxBlockDistance = phylogenyMaxBlockDistance;
                params.numTrees = phylogenyNumTrees;
                params.ignoreUnalignedBases = 1;
                params.onlyIncludeCompleteFeatureBlocks = 0;
                params.doSplitsWithSupportHigherThanThisAllAtOnce = phylogenyDoSplitsWithSupportHigherThanThisAllAtOnce;
                params.numTreeBuildingThreads = numTreeBuildingThreads;

                assert(params.numTreeBuildingThreads >= 1);

                stCaf_buildTreesToRemoveAncientHomologies(
                    threadSet, phylogenyHomologyUnitType, threadStrings, outgroupThreads, flower, &params,
                    debugFileName == NULL ? NULL : stString_print("%s-phylogeny", debugFileName), referenceEventHeader);
                stHash_destruct(threadStrings);
                st_logDebug("Finished building trees\n");

                if (removeRecoverableChains) {
                    // We melt recoverable chains after splitting, as
                    // well as before, to alleviate coverage loss
                    // caused by bad splits.
                    stCaf_meltRecoverableChains(flower, threadSet, breakChainsAtReverseTandems, maximumMedianSequenceLengthBetweenLinkedEnds, recoverableChainsFilter, maxRecoverableChainsIterations, maxRecoverableChainLength);
                }

                // Enforce the block constraints on minimum degree,
                // etc. after splitting.
                stCaf_melt(flower, threadSet, blockFilterFn, 0, 0, 0, INT64_MAX);
            }

            //Sort out case when we allow blocks of degree 1
            if (minimumDegree < 2) {
                st_logDebug("Creating degree 1 blocks\n");
                stCaf_makeDegreeOneBlocks(threadSet);
                stCaf_melt(flower, threadSet, blockFilterFn, blockTrim, 0, 0, INT64_MAX);
            } else if (maximumAdjacencyComponentSizeRatio < INT64_MAX) { //Deal with giant components
                st_logDebug("Breaking up components greedily\n");
                stCaf_breakupComponentsGreedily(threadSet, maximumAdjacencyComponentSizeRatio);
            }

            //Finish up
            stCaf_finish(flower, threadSet, chainLengthForBigFlower, longChain, minLengthForChromosome,
                    proportionOfUnalignedBasesForNewChromosome); //Flower is then destroyed at this point.
            st_logInfo("Ran the cactus core script\n");

            //Cleanup
            stPinchThreadSet_destruct(threadSet);
            stPinchIterator_destruct(pinchIterator);
            stSet_destruct(outgroupThreads);

            if (alignmentsList != NULL) {
                stList_destruct(alignmentsList);
            }
            st_logInfo("Cleaned up from main loop\n");
        } else {
            st_logInfo("We've already built blocks / alignments for this flower\n");
        }
    }
    stList_destruct(flowers);
    if (tempFile1 != NULL) {
        st_system("rm %s", tempFile1);
    }

    if (constraintsFile != NULL) {
        stPinchIterator_destruct(pinchIteratorForConstraints);
    }

    ///////////////////////////////////////////////////////////////////////////
    // Write the flower to disk.
    ///////////////////////////////////////////////////////////////////////////
    st_logDebug("Writing the flowers to disk\n");
    cactusDisk_write(cactusDisk);
    st_logInfo("Updated the flower on disk and %" PRIi64 " seconds have elapsed\n", time(NULL) - startTime);

    ///////////////////////////////////////////////////////////////////////////
    // Clean up.
    ///////////////////////////////////////////////////////////////////////////

    cactusDisk_destruct(cactusDisk);
}