]> git.donarmstrong.com Git - mothur.git/blobdiff - maligner.cpp
working on slayer bug
[mothur.git] / maligner.cpp
index b393194bbc3a4c89fd2ae8ea3f7f7ba67f84b927..c5cc83a4d216b9c5a89eb6f60de8a15e5906d06c 100644 (file)
@@ -8,12 +8,23 @@
  */
 
 #include "maligner.h"
-#include "database.hpp"
+#include "kmerdb.hpp"
 #include "blastdb.hpp"
 
 /***********************************************************************/
-Maligner::Maligner(vector<Sequence*> temp, int num, int match, int misMatch, float div, int ms, int minCov, string mode) :
-               db(temp), numWanted(num), matchScore(match), misMatchPenalty(misMatch), minDivR(div), minSimilarity(ms), minCoverage(minCov), searchMethod(mode) {}
+Maligner::Maligner(vector<Sequence*> temp, int num, int match, int misMatch, float div, int ms, int minCov, string mode, Database* dataLeft, Database* dataRight) :
+               db(temp), numWanted(num), matchScore(match), misMatchPenalty(misMatch), minDivR(div), minSimilarity(ms), minCoverage(minCov), searchMethod(mode), databaseLeft(dataLeft), databaseRight(dataRight) { 
+                       
+                       
+                       m = MothurOut::getInstance(); 
+                       
+//                     cout << matchScore << '\t' << misMatchPenalty << endl;
+//                     
+//                     matchScore = 1;
+//                     misMatchPenalty = -1;
+                       
+               }
+
 /***********************************************************************/
 string Maligner::getResults(Sequence* q, DeCalculator* decalc) {
        try {
@@ -25,15 +36,21 @@ string Maligner::getResults(Sequence* q, DeCalculator* decalc) {
                
                string chimera;
                
-               if (searchMethod != "blast") {
+               if (searchMethod == "distance") {
                        //find closest seqs to query in template - returns copies of seqs so trim does not destroy - remember to deallocate
-                       refSeqs = decalc->findClosest(query, db, numWanted);
-               }else{
-                       refSeqs = getBlastSeqs(query, numWanted);
-               }
+                       refSeqs = decalc->findClosest(query, db, numWanted, indexes);
+               }else if (searchMethod == "blast")  {
+                       refSeqs = getBlastSeqs(query, numWanted); //fills indexes
+               }else if (searchMethod == "kmer") {
+                       refSeqs = getKmerSeqs(query, numWanted); //fills indexes
+               }else { m->mothurOut("not valid search."); exit(1);  } //should never get here
+               
+               if (m->control_pressed) { return chimera;  }
                
                refSeqs = minCoverageFilter(refSeqs);
                
+               
+               
                if (refSeqs.size() < 2)  { 
                        for (int i = 0; i < refSeqs.size(); i++) {  delete refSeqs[i];  }
                        percentIdenticalQueryChimera = 0.0;
@@ -41,19 +58,21 @@ string Maligner::getResults(Sequence* q, DeCalculator* decalc) {
                }
                
                int chimeraPenalty = computeChimeraPenalty();
-       
+               //cout << "chimeraPenalty = " << chimeraPenalty << endl;
                //fills outputResults
                chimera = chimeraMaligner(chimeraPenalty, decalc);
                
+               if (m->control_pressed) { return chimera;  }
                                
                //free memory
                delete query;
+
                for (int i = 0; i < refSeqs.size(); i++) {  delete refSeqs[i];  }
                
                return chimera;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "getResults");
+               m->errorOut(e, "Maligner", "getResults");
                exit(1);
        }
 }
@@ -66,31 +85,38 @@ string Maligner::chimeraMaligner(int chimeraPenalty, DeCalculator* decalc) {
                //trims seqs to first non gap char in all seqs and last non gap char in all seqs
                spotMap = decalc->trimSeqs(query, refSeqs);
                
+               //you trimmed the whole sequence, skip
+               if (query->getAligned() == "") { return "no"; }
+
                vector<Sequence*> temp = refSeqs;
                temp.push_back(query);
-               
+                       
                verticalFilter(temp);
+               
+               //for (int i = 0; i < refSeqs.size(); i++) { cout << refSeqs[i]->getName() << endl << refSeqs[i]->getAligned() << endl; }
 
                vector< vector<score_struct> > matrix = buildScoreMatrix(query->getAligned().length(), refSeqs.size()); //builds and initializes
                
-               fillScoreMatrix(matrix, refSeqs, chimeraPenalty);
-               
-               vector<score_struct> path = extractHighestPath(matrix);
+               if (m->control_pressed) { return chimera;  }
                
-               vector<trace_struct> trace = mapTraceRegionsToAlignment(path, refSeqs);
+               fillScoreMatrix(matrix, refSeqs, chimeraPenalty);
+       
+               vector<trace_struct> trace = extractHighestPath(matrix);
+                               
+               //cout << "traces\n";
+               //for(int i=0;i<trace.size();i++){
+               //      cout << trace[i].col << '\t' << trace[i].oldCol << '\t' << refSeqs[trace[i].row]->getName() << endl;
+               //}
                
                if (trace.size() > 1) {         chimera = "yes";        }
-               else { chimera = "no";  }
-               
-               int traceStart = path[0].col;
-               int traceEnd = path[path.size()-1].col;
+               else { chimera = "no";  return chimera; }
                
+               int traceStart = trace[0].col;
+               int traceEnd = trace[trace.size()-1].oldCol;    
                string queryInRange = query->getAligned();
                queryInRange = queryInRange.substr(traceStart, (traceEnd-traceStart+1));
-       
-               string chimeraSeq = constructChimericSeq(trace, refSeqs);
                
-               percentIdenticalQueryChimera = computePercentID(queryInRange, chimeraSeq);
+               if (m->control_pressed) { return chimera;  }
                
                //save output results
                for (int i = 0; i < trace.size(); i++) {
@@ -101,6 +127,7 @@ string Maligner::chimeraMaligner(int chimeraPenalty, DeCalculator* decalc) {
                        results temp;
                        
                        temp.parent = refSeqs[seqIndex]->getName();
+                       temp.parentAligned = db[indexes[seqIndex]]->getAligned();
                        temp.nastRegionStart = spotMap[regionStart];
                        temp.nastRegionEnd = spotMap[regionEnd];
                        temp.regionStart = regionStart;
@@ -111,7 +138,7 @@ string Maligner::chimeraMaligner(int chimeraPenalty, DeCalculator* decalc) {
                        
                        temp.queryToParent = computePercentID(queryInRange, parentInRange);
                        temp.divR = (percentIdenticalQueryChimera / temp.queryToParent);
-                       
+
                        string queryInRegion = query->getAligned();
                        queryInRegion = queryInRegion.substr(regionStart, (regionEnd-regionStart+1));
                        
@@ -122,11 +149,11 @@ string Maligner::chimeraMaligner(int chimeraPenalty, DeCalculator* decalc) {
                
                        outputResults.push_back(temp);
                }
-
+               
                return chimera;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "chimeraMaligner");
+               m->errorOut(e, "Maligner", "chimeraMaligner");
                exit(1);
        }
 }
@@ -162,13 +189,15 @@ vector<Sequence*> Maligner::minCoverageFilter(vector<Sequence*> ref){
                        //if coverage above minimum
                        if (coverage > minCoverage) {
                                newRefs.push_back(ref[i]);
+                       }else {
+                               delete ref[i];
                        }
                }
                
                return newRefs;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "minCoverageFilter");
+               m->errorOut(e, "Maligner", "minCoverageFilter");
                exit(1);
        }
 }
@@ -178,14 +207,16 @@ int Maligner::computeChimeraPenalty() {
        try {
                
                int numAllowable = ((1.0 - (1.0/minDivR)) * query->getNumBases());
-       
+
+//             if(numAllowable < 1){   numAllowable = 1;       }
+               
                int penalty = int(numAllowable + 1) * misMatchPenalty;
-                                                                                        
+
                return penalty;
 
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "computeChimeraPenalty");
+               m->errorOut(e, "Maligner", "computeChimeraPenalty");
                exit(1);
        }
 }
@@ -237,7 +268,7 @@ void Maligner::verticalFilter(vector<Sequence*> seqs) {
                spotMap = newMap;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "verticalFilter");
+               m->errorOut(e, "Maligner", "verticalFilter");
                exit(1);
        }
 }
@@ -245,9 +276,9 @@ void Maligner::verticalFilter(vector<Sequence*> seqs) {
 vector< vector<score_struct> > Maligner::buildScoreMatrix(int cols, int rows) {
        try{
                
-               vector< vector<score_struct> > m; m.resize(rows);
+               vector< vector<score_struct> > m(rows);
                
-               for (int i = 0; i < m.size(); i++) {
+               for (int i = 0; i < rows; i++) {
                        for (int j = 0; j < cols; j++) {
                                
                                //initialize each cell
@@ -264,12 +295,14 @@ vector< vector<score_struct> > Maligner::buildScoreMatrix(int cols, int rows) {
                return m;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "buildScoreMatrix");
+               m->errorOut(e, "Maligner", "buildScoreMatrix");
                exit(1);
        }
 }
+
 //***************************************************************************************************************
-void Maligner::fillScoreMatrix(vector<vector<score_struct> >& m, vector<Sequence*> seqs, int penalty) {
+
+void Maligner::fillScoreMatrix(vector<vector<score_struct> >& ms, vector<Sequence*> seqs, int penalty) {
        try{
                
                //get matrix dimensions
@@ -283,11 +316,14 @@ void Maligner::fillScoreMatrix(vector<vector<score_struct> >& m, vector<Sequence
                        
                        //are you both gaps?
                        if ((!isalpha(queryAligned[0])) && (!isalpha(subjectAligned[0]))) {
-                               m[i][0].score = 0;
-                       }else if (queryAligned[0] == subjectAligned[0]) {
-                               m[i][0].score = matchScore;
+                               ms[i][0].score = 0;
+//                             ms[i][0].mismatches = 0;
+                       }else if (queryAligned[0] == subjectAligned[0])  { //|| subjectAligned[0] == 'N')
+                               ms[i][0].score = matchScore;
+//                             ms[i][0].mismatches = 0;
                        }else{
-                               m[i][0].score = 0;
+                               ms[i][0].score = 0;
+//                             ms[i][0].mismatches = 1;
                        }
                }
                
@@ -303,85 +339,160 @@ void Maligner::fillScoreMatrix(vector<vector<score_struct> >& m, vector<Sequence
                                if ((!isalpha(queryAligned[j])) && (!isalpha(subjectAligned[j]))) {
                                        //leave the same
                                }else if ((toupper(queryAligned[j]) == 'N') || (toupper(subjectAligned[j]) == 'N')) {
+                                       //matchMisMatchScore = matchScore;
                                        //leave the same
                                }else if (queryAligned[j] == subjectAligned[j]) {
                                        matchMisMatchScore = matchScore;
+//                                     ms[i][j].mismatches = ms[i][j-1].mismatches;
                                }else if (queryAligned[j] != subjectAligned[j]) {
                                        matchMisMatchScore = misMatchPenalty;
+//                                     ms[i][j].mismatches = ms[i][j-1].mismatches + 1;
                                }
                                
                                //compute score based on previous columns scores
                                for (int prevIndex = 0; prevIndex < numRows; prevIndex++) { //iterate through rows
                                        
-                                       int sumScore = matchMisMatchScore + m[prevIndex][j-1].score;
+                                       int sumScore = matchMisMatchScore + ms[prevIndex][j-1].score;
                                        
                                        //you are not at yourself
                                        if (prevIndex != i) {   sumScore += penalty;    }
                                        if (sumScore < 0)       {       sumScore = 0;                   }
                                        
-                                       if (sumScore > m[i][j].score) {
-                                               m[i][j].score = sumScore;
-                                               m[i][j].prev = prevIndex;
+                                       if (sumScore > ms[i][j].score) {
+                                               ms[i][j].score = sumScore;
+                                               ms[i][j].prev = prevIndex;
                                        }
                                }
                        }
                }
                
+       /*      for(int i=0;i<numRows;i++){
+                       cout << seqs[i]->getName();
+                       for(int j=0;j<numCols;j++){
+                               cout << '\t' << ms[i][j].mismatches;
+                       }
+                       cout << endl;
+               }
+               cout << endl;*/
+               /*cout << numRows << '\t' << numCols << endl;
+               for(int i=0;i<numRows;i++){
+                       cout << seqs[i]->getName() << endl << seqs[i]->getAligned() << endl << endl;
+                       if ((seqs[i]->getName() == "S000003470") || (seqs[i]->getName() == "S000383265") || (seqs[i]->getName() == "7000004128191054")) {
+                       for(int j=0;j<numCols;j++){
+                               cout << '\t' << ms[i][j].score;
+                       }
+                       cout << endl;
+                       }
+               }
+               cout << endl;*/
+               /*for(int i=0;i<numRows;i++){
+                       cout << seqs[i]->getName();
+                       for(int j=0;j<numCols;j++){
+                               cout << '\t' << ms[i][j].prev;
+                       }
+                       cout << endl;
+               }*/
+               
+               
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "fillScoreMatrix");
+               m->errorOut(e, "Maligner", "fillScoreMatrix");
                exit(1);
        }
 }
+
 //***************************************************************************************************************
-vector<score_struct> Maligner::extractHighestPath(vector<vector<score_struct> > m) {
+
+vector<trace_struct> Maligner::extractHighestPath(vector<vector<score_struct> > ms) {
        try {
        
+               
                //get matrix dimensions
                int numCols = query->getAligned().length();
-               int numRows = m.size();
+               int numRows = ms.size();
        
        
                //find highest score scoring matrix
-               score_struct highestStruct;
+               vector<score_struct> highestStruct;
                int highestScore = 0;
                
                for (int i = 0; i < numRows; i++) {
                        for (int j = 0; j < numCols; j++) {
-                               if (m[i][j].score > highestScore) {
-                                       highestScore = m[i][j].score;
-                                       highestStruct = m[i][j];
+                               if (ms[i][j].score > highestScore) {
+                                       highestScore = ms[i][j].score;
+                                       highestStruct.resize(0);
+                                       highestStruct.push_back(ms[i][j]);
+                               }
+                               else if(ms[i][j].score == highestScore){
+                                       highestStruct.push_back(ms[i][j]);
                                }
                        }
                }
-                               
-               vector<score_struct> path;
+                       
+               //cout << endl << highestScore << '\t' << highestStruct.size() << '\t' << highestStruct[0].row << endl; 
                
-               int rowIndex = highestStruct.row;
-               int pos = highestStruct.col;
-               int score = highestStruct.score;
+               vector<trace_struct> maxTrace;
+               double maxPercentIdenticalQueryAntiChimera = 0;
                
-               while (pos >= 0 && score > 0) {
-                       score_struct temp = m[rowIndex][pos];
-                       score = temp.score;
+               for(int i=0;i<highestStruct.size();i++){
+                       
+                       vector<score_struct> path;
+
+                       int rowIndex = highestStruct[i].row;
+                       int pos = highestStruct[i].col;
+                       int score = highestStruct[i].score;
+                                       
+                       while (pos >= 0 && score > 0) {
+                               score_struct temp = ms[rowIndex][pos];
+                               score = temp.score;
+                               
+                               if (score > 0) {        path.push_back(temp);   }
+                               
+                               rowIndex = temp.prev;
+                               pos--;
+                       }
+
+                       reverse(path.begin(), path.end());
+
+                       vector<trace_struct> trace = mapTraceRegionsToAlignment(path, refSeqs);
+               
+                       //cout << "traces\n";
+                       //for(int j=0;j<trace.size();j++){
+                       //      cout << trace[j].col << '\t' << trace[j].oldCol << '\t' << refSeqs[trace[j].row]->getName() << endl;
+                       //}
+                                               
+                       int traceStart = path[0].col;
+                       int traceEnd = path[path.size()-1].col; 
+//                     cout << "traceStart/End\t" << traceStart << '\t' << traceEnd << endl;
                        
-                       if (score > 0) {        path.push_back(temp);   }
+                       string queryInRange = query->getAligned();
+                       queryInRange = queryInRange.substr(traceStart, (traceEnd-traceStart+1));
+//                     cout << "here" << endl;
+                       string chimeraSeq = constructChimericSeq(trace, refSeqs);
+                       string antiChimeraSeq = constructAntiChimericSeq(trace, refSeqs);
+               
+                       percentIdenticalQueryChimera = computePercentID(queryInRange, chimeraSeq);                      
+                       double percentIdenticalQueryAntiChimera = computePercentID(queryInRange, antiChimeraSeq);
+//                     cout << i << '\t' << percentIdenticalQueryChimera << '\t' << percentIdenticalQueryAntiChimera << endl;
                        
-                       rowIndex = temp.prev;
-                       pos--;
+                       if(percentIdenticalQueryAntiChimera > maxPercentIdenticalQueryAntiChimera){
+                               maxPercentIdenticalQueryAntiChimera = percentIdenticalQueryAntiChimera;
+                               maxTrace = trace;
+                       }
                }
-
-               reverse(path.begin(), path.end());
+//             cout << maxPercentIdenticalQueryAntiChimera << endl;
+               return maxTrace;
        
-               return path;
                
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "extractHighestPath");
+               m->errorOut(e, "Maligner", "extractHighestPath");
                exit(1);
        }
 }
+
 //***************************************************************************************************************
+
 vector<trace_struct> Maligner::mapTraceRegionsToAlignment(vector<score_struct> path, vector<Sequence*> seqs) {
        try {
                vector<trace_struct> trace;
@@ -416,20 +527,31 @@ vector<trace_struct> Maligner::mapTraceRegionsToAlignment(vector<score_struct> p
                temp.row = region_index;
                trace.push_back(temp);
 
+//             cout << endl;
+//             cout << trace.size() << endl;
+//             for(int i=0;i<trace.size();i++){
+//                     cout << seqs[trace[i].row]->getName() << endl;
+//             }
+//             cout << endl;
+               
                return trace;
                
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "mapTraceRegionsToAlignment");
+               m->errorOut(e, "Maligner", "mapTraceRegionsToAlignment");
                exit(1);
        }
 }
+
 //***************************************************************************************************************
+
 string Maligner::constructChimericSeq(vector<trace_struct> trace, vector<Sequence*> seqs) {
        try {
                string chimera = "";
                
                for (int i = 0; i < trace.size(); i++) {
+//                     cout << i << '\t' << trace[i].row << '\t' << trace[i].col << '\t' << trace[i].oldCol << endl;
+                       
                        string seqAlign = seqs[trace[i].row]->getAligned();
                        seqAlign = seqAlign.substr(trace[i].col, (trace[i].oldCol-trace[i].col+1));
                        chimera += seqAlign;
@@ -438,18 +560,43 @@ string Maligner::constructChimericSeq(vector<trace_struct> trace, vector<Sequenc
                return chimera;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "constructChimericSeq");
+               m->errorOut(e, "Maligner", "constructChimericSeq");
+               exit(1);
+       }
+}
+
+//***************************************************************************************************************
+
+string Maligner::constructAntiChimericSeq(vector<trace_struct> trace, vector<Sequence*> seqs) {
+       try {
+               string antiChimera = "";
+               
+               for (int i = 0; i < trace.size(); i++) {
+//                     cout << i << '\t' << (trace.size() - i - 1) << '\t' << trace[i].row << '\t' << trace[i].col << '\t' << trace[i].oldCol << endl;
+                       
+                       int oppositeIndex = trace.size() - i - 1;
+                       
+                       string seqAlign = seqs[trace[oppositeIndex].row]->getAligned();
+                       seqAlign = seqAlign.substr(trace[i].col, (trace[i].oldCol-trace[i].col+1));
+                       antiChimera += seqAlign;
+               }
+               
+               return antiChimera;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "Maligner", "constructChimericSeq");
                exit(1);
        }
 }
+
 //***************************************************************************************************************
 float Maligner::computePercentID(string queryAlign, string chimera) {
        try {
        
                if (queryAlign.length() != chimera.length()) {
-                       mothurOut("Error, alignment strings are of different lengths: "); mothurOutEndLine();
-                       mothurOut(toString(queryAlign.length())); mothurOutEndLine(); mothurOutEndLine(); mothurOutEndLine(); mothurOutEndLine();
-                       mothurOut(toString(chimera.length())); mothurOutEndLine();
+                       m->mothurOut("Error, alignment strings are of different lengths: "); m->mothurOutEndLine();
+                       m->mothurOut(toString(queryAlign.length())); m->mothurOutEndLine(); m->mothurOutEndLine(); m->mothurOutEndLine(); m->mothurOutEndLine();
+                       m->mothurOut(toString(chimera.length())); m->mothurOutEndLine();
                        return -1.0;
                }
 
@@ -474,18 +621,160 @@ float Maligner::computePercentID(string queryAlign, string chimera) {
                
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "computePercentID");
+               m->errorOut(e, "Maligner", "computePercentID");
                exit(1);
        }
 }
 //***************************************************************************************************************
 vector<Sequence*> Maligner::getBlastSeqs(Sequence* q, int num) {
        try {   
-               //generate blastdb
-               Database* database = new BlastDB(-2.0, -1.0, matchScore, misMatchPenalty);
-               for (int i = 0; i < db.size(); i++) {   database->addSequence(*db[i]);  }
-               database->generateDB();
-               database->setNumSeqs(db.size());
+               indexes.clear();
+               vector<Sequence*> refResults;
+                               
+               //get parts of query
+               string queryUnAligned = q->getUnaligned();
+               string leftQuery = queryUnAligned.substr(0, int(queryUnAligned.length() * 0.33)); //first 1/3 of the sequence
+               string rightQuery = queryUnAligned.substr(int(queryUnAligned.length() * 0.66)); //last 1/3 of the sequence
+               
+               Sequence* queryLeft = new Sequence(q->getName()+"left", leftQuery);
+               Sequence* queryRight = new Sequence(q->getName()+"right", rightQuery);
+
+               vector<int> tempIndexesLeft = databaseLeft->findClosestMegaBlast(queryLeft, num+1);
+               vector<float> leftScores = databaseLeft->getSearchScores();
+               vector<int> tempIndexesRight = databaseLeft->findClosestMegaBlast(queryRight, num+1);
+               vector<float> rightScores = databaseLeft->getSearchScores();
+
+               //if ((tempIndexesRight.size() == 0) && (tempIndexesLeft.size() == 0))  {  m->mothurOut("megablast returned " + toString(tempIndexesRight.size()) + " results for the right end, and " + toString(tempIndexesLeft.size()) + " for the left end. Needed " + toString(num+1) + ". Unable to process sequence " + q->getName()); m->mothurOutEndLine(); return refResults; }
+               
+               vector<int> smaller;
+               vector<float> smallerScores;
+               vector<int> larger;
+               vector<float> largerScores;
+               
+               if (tempIndexesRight.size() < tempIndexesLeft.size()) { smaller = tempIndexesRight; smallerScores = rightScores; larger = tempIndexesLeft; largerScores = leftScores; }
+               else { smaller = tempIndexesLeft; smallerScores = leftScores; larger = tempIndexesRight; largerScores = rightScores; } 
+               
+               //for (int i = 0; i < smaller.size(); i++) { cout << "smaller = " << smaller[i] << '\t' << smallerScores[i] << endl; }
+               //cout << endl;
+               //for (int i = 0; i < larger.size(); i++) { cout << "larger = " << larger[i] << '\t' << largerScores[i] << endl; }
+               
+               //merge results         
+               map<int, int> seen;
+               map<int, int>::iterator it;
+               float lastSmaller = smallerScores[0];
+               float lastLarger = largerScores[0];
+               int lasti = 0;
+               vector<int> mergedResults;
+               for (int i = 0; i < smaller.size(); i++) {
+                       //add left if you havent already
+                       it = seen.find(smaller[i]);
+                       if (it == seen.end()) {  
+                               mergedResults.push_back(smaller[i]);
+                               seen[smaller[i]] = smaller[i];
+                               lastSmaller = smallerScores[i];
+                       }
+
+                       //add right if you havent already
+                       it = seen.find(larger[i]);
+                       if (it == seen.end()) {  
+                               mergedResults.push_back(larger[i]);
+                               seen[larger[i]] = larger[i];
+                               lastLarger = largerScores[i];
+                       }
+                       
+                       lasti = i;
+                       //if (mergedResults.size() > num) { break; }
+               }
+               
+               //save lasti for smaller ties below
+               /*lasti++;
+               int iSmaller = lasti;
+               
+               if (!(mergedResults.size() > num)) { //if we still need more results.  
+                       for (int i = smaller.size(); i < larger.size(); i++) {
+                               it = seen.find(larger[i]);
+                               if (it == seen.end()) {  
+                                       mergedResults.push_back(larger[i]);
+                                       seen[larger[i]] = larger[i];
+                                       lastLarger = largerScores[i];
+                               }
+                               
+                               lasti = i;
+                               if (mergedResults.size() > num) {  break; }
+                       }
+               }
+               
+               
+               //add in any ties from smaller
+               while (iSmaller < smaller.size()) {
+                       if (smallerScores[iSmaller] == lastSmaller) {
+                               it = seen.find(smaller[iSmaller]);
+                               
+                               if (it == seen.end()) {  
+                                       mergedResults.push_back(smaller[iSmaller]);
+                                       seen[smaller[iSmaller]] = smaller[iSmaller];
+                               }
+                       }
+                       else { break; }
+                       iSmaller++;                     
+               }
+               
+               lasti++;
+               //add in any ties from larger
+               while (lasti < larger.size()) {
+                       if (largerScores[lasti] == lastLarger) { //is it a tie
+                               it = seen.find(larger[lasti]);
+                               
+                               if (it == seen.end()) {  //we haven't already seen it
+                                       mergedResults.push_back(larger[lasti]);
+                                       seen[larger[lasti]] = larger[lasti];
+                               }
+                       }
+                       else { break; }
+                       lasti++;                        
+               }
+               */
+               
+               for (int i = smaller.size(); i < larger.size(); i++) {
+                       //add right if you havent already
+                       it = seen.find(larger[i]);
+                       if (it == seen.end()) {  
+                               mergedResults.push_back(larger[i]);
+                               seen[larger[i]] = larger[i];
+                               lastLarger = largerScores[i];
+                       }
+               }
+               numWanted = mergedResults.size();
+               
+               if (mergedResults.size() < numWanted) { numWanted = mergedResults.size(); }
+//cout << q->getName() << " merged results size = " << mergedResults.size() << '\t' << "numwanted = " << numWanted <<  endl;           
+               for (int i = 0; i < numWanted; i++) {
+//cout << db[mergedResults[i]]->getName()  << '\t' << mergedResults[i] << endl;        
+                       
+                       if (db[mergedResults[i]]->getName() != q->getName()) { 
+                               Sequence* temp = new Sequence(db[mergedResults[i]]->getName(), db[mergedResults[i]]->getAligned());
+                               refResults.push_back(temp);
+                               indexes.push_back(mergedResults[i]);
+                               //cout << db[mergedResults[i]]->getName() << endl;
+                       }
+                       
+//cout << mergedResults[i] << endl;
+               }
+//cout << "done " << q->getName()  << endl;            
+               delete queryRight;
+               delete queryLeft;
+                       
+               return refResults;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "Maligner", "getBlastSeqs");
+               exit(1);
+       }
+}
+//***************************************************************************************************************
+vector<Sequence*> Maligner::getKmerSeqs(Sequence* q, int num) {
+       try {   
+               indexes.clear();
                
                //get parts of query
                string queryUnAligned = q->getUnaligned();
@@ -495,13 +784,17 @@ vector<Sequence*> Maligner::getBlastSeqs(Sequence* q, int num) {
                Sequence* queryLeft = new Sequence(q->getName(), leftQuery);
                Sequence* queryRight = new Sequence(q->getName(), rightQuery);
                
-               vector<int> tempIndexesRight = database->findClosestMegaBlast(queryRight, num+1);
-               vector<int> tempIndexesLeft = database->findClosestMegaBlast(queryLeft, num+1);
+               vector<int> tempIndexesLeft = databaseLeft->findClosestSequences(queryLeft, numWanted);
+               vector<int> tempIndexesRight = databaseRight->findClosestSequences(queryRight, numWanted);
+               vector<float> scoresLeft = databaseLeft->getSearchScores();
+               vector<float> scoresRight = databaseRight->getSearchScores();
                
                //merge results         
                map<int, int> seen;
                map<int, int>::iterator it;
-               
+               float lastRight = scoresRight[0];
+               float lastLeft = scoresLeft[0];
+               //int lasti = 0;
                vector<int> mergedResults;
                for (int i = 0; i < tempIndexesLeft.size(); i++) {
                        //add left if you havent already
@@ -509,6 +802,7 @@ vector<Sequence*> Maligner::getBlastSeqs(Sequence* q, int num) {
                        if (it == seen.end()) {  
                                mergedResults.push_back(tempIndexesLeft[i]);
                                seen[tempIndexesLeft[i]] = tempIndexesLeft[i];
+                               lastLeft = scoresLeft[i];
                        }
 
                        //add right if you havent already
@@ -516,27 +810,72 @@ vector<Sequence*> Maligner::getBlastSeqs(Sequence* q, int num) {
                        if (it == seen.end()) {  
                                mergedResults.push_back(tempIndexesRight[i]);
                                seen[tempIndexesRight[i]] = tempIndexesRight[i];
+                               lastRight = scoresRight[i];
+                       }
+                       
+                       //if (mergedResults.size() > numWanted) { lasti = i; break; } //you have enough results
+               }
+               
+               //add in sequences with same distance as last sequence added
+               /*lasti++;
+               int i = lasti;
+               while (i < tempIndexesLeft.size()) {  
+                       if (scoresLeft[i] == lastLeft) {
+                               it = seen.find(tempIndexesLeft[i]);
+                               
+                               if (it == seen.end()) {  
+                                       mergedResults.push_back(tempIndexesLeft[i]);
+                                       seen[tempIndexesLeft[i]] = tempIndexesLeft[i];
+                               }
                        }
+                       else { break; }
+                       i++;
+               }
+               
+               //              cout << "lastRight\t" << lastRight << endl;
+               //add in sequences with same distance as last sequence added
+               i = lasti;
+               while (i < tempIndexesRight.size()) {  
+                       if (scoresRight[i] == lastRight) {
+                               it = seen.find(tempIndexesRight[i]);
+                               
+                               if (it == seen.end()) {  
+                                       mergedResults.push_back(tempIndexesRight[i]);
+                                       seen[tempIndexesRight[i]] = tempIndexesRight[i];
+                               }
+                       }
+                       else { break; }
+                       i++;
+               }*/
+               
+               numWanted = mergedResults.size();
+               
+               if (numWanted > mergedResults.size()) { 
+                       //m->mothurOut("numwanted is larger than the number of template sequences, adjusting numwanted."); m->mothurOutEndLine(); 
+                       numWanted = mergedResults.size();
                }
                
                
+//cout << q->getName() << endl;                
                vector<Sequence*> refResults;
                for (int i = 0; i < numWanted; i++) {
-                       Sequence* temp = new Sequence(db[mergedResults[i]]->getName(), db[mergedResults[i]]->getAligned());
-                       refResults.push_back(temp);
+//cout << db[mergedResults[i]]->getName() << endl;     
+                       if (db[mergedResults[i]]->getName() != q->getName()) { 
+                               Sequence* temp = new Sequence(db[mergedResults[i]]->getName(), db[mergedResults[i]]->getAligned());
+                               refResults.push_back(temp);
+                               indexes.push_back(mergedResults[i]);
+                       }
                }
-               
+//cout << endl;                
                delete queryRight;
                delete queryLeft;
-               delete database;
                
                return refResults;
        }
        catch(exception& e) {
-               errorOut(e, "Maligner", "getBlastSeqs");
+               m->errorOut(e, "Maligner", "getKmerSeqs");
                exit(1);
        }
 }
-
 //***************************************************************************************************************