]> git.donarmstrong.com Git - mothur.git/blobdiff - trimoligos.cpp
added merge.taxsummary command. added column format to dist.shared.
[mothur.git] / trimoligos.cpp
index c35971a676f1c9af3de404bb5f0a4ca707a82f1f..a53f4e85388f0f7bbb6fda40c2074b4f555e29d3 100644 (file)
@@ -144,6 +144,24 @@ TrimOligos::TrimOligos(int p, int b, map<string, int> pr, map<string, int> br, v
                barcodes = br;
                primers = pr;
                revPrimer = r;
+        
+        maxFBarcodeLength = 0;
+        for(map<string,int>::iterator it=barcodes.begin();it!=barcodes.end();it++){
+            string oligo = it->first;
+            if(oligo.length() > maxFBarcodeLength){
+                maxFBarcodeLength = oligo.length();
+            }
+        }
+        maxRBarcodeLength = maxFBarcodeLength;
+        
+        maxFPrimerLength = 0;
+        for(map<string,int>::iterator it=primers.begin();it!=primers.end();it++){
+            string oligo = it->first;
+            if(oligo.length() > maxFPrimerLength){
+                maxFPrimerLength = oligo.length();
+            }
+        }
+        maxRPrimerLength = maxFPrimerLength;
        }
        catch(exception& e) {
                m->errorOut(e, "TrimOligos", "TrimOligos");
@@ -152,7 +170,135 @@ TrimOligos::TrimOligos(int p, int b, map<string, int> pr, map<string, int> br, v
 }
 /********************************************************************/
 TrimOligos::~TrimOligos() {}
+//********************************************************************/
+bool TrimOligos::findForward(Sequence& seq, int& primerStart, int& primerEnd){
+       try {
+               
+               string rawSequence = seq.getUnaligned();
+               
+               for(map<string,int>::iterator it=primers.begin();it!=primers.end();it++){
+                       string oligo = it->first;
+                       
+                       if(rawSequence.length() < oligo.length()) {  break;  }
+                       
+                       //search for primer
+            int olength = oligo.length();
+            for (int j = 0; j < rawSequence.length()-olength; j++){
+                if (m->control_pressed) {  primerStart = 0; primerEnd = 0; return false; }
+                string rawChunk = rawSequence.substr(j, olength);
+                if(compareDNASeq(oligo, rawChunk)) {
+                    primerStart = j;
+                    primerEnd = primerStart + olength;
+                    return true;
+                }
+                
+            }
+        }      
+               
+        primerStart = 0; primerEnd = 0;
+        //if you don't want to allow for diffs
+               if (pdiffs == 0) { return false;  }
+               else { //try aligning and see if you can find it
+                                               
+                       //can you find the barcode
+                       int minDiff = 1e6;
+                       int minCount = 1;
+                       
+            Alignment* alignment;
+            if (primers.size() > 0) { alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxFPrimerLength+pdiffs+1));  }
+            else{ alignment = NULL; } 
+            
+                       for(map<string,int>::iterator it=primers.begin();it!=primers.end();it++){
+                
+                string prim = it->first;
+                //search for primer
+                int olength = prim.length();
+                if (rawSequence.length() < olength) {} //ignore primers too long for this seq
+                else{
+                    for (int j = 0; j < rawSequence.length()-olength; j++){
+                        
+                        string oligo = it->first;
+
+                        if (m->control_pressed) {  primerStart = 0; primerEnd = 0; return false; }
+                               
+                        string rawChunk = rawSequence.substr(j, olength+pdiffs);
+                        
+                        //use needleman to align first primer.length()+numdiffs of sequence to each barcode
+                        alignment->align(oligo, rawChunk);
+                        oligo = alignment->getSeqAAln();
+                        string temp = alignment->getSeqBAln();
+                               
+                        int alnLength = oligo.length();
+                               
+                        for(int i=oligo.length()-1;i>=0;i--){
+                            if(oligo[i] != '-'){       alnLength = i+1;        break;  }
+                        }
+                        oligo = oligo.substr(0,alnLength);
+                        temp = temp.substr(0,alnLength);
+                               
+                        int numDiff = countDiffs(oligo, temp);
+                               
+                        if(numDiff < minDiff){
+                            minDiff = numDiff;
+                            minCount = 1;
+                            primerStart = j;
+                            primerEnd = primerStart + alnLength;
+                        }else if(numDiff == minDiff){ minCount++; }
+                    }
+                }
+                       }
+                       
+            if (alignment != NULL) {  delete alignment;  }
+            
+                       if(minDiff > pdiffs)    {       primerStart = 0; primerEnd = 0; return false;           }       //no good matches
+                       else if(minCount > 1)   {       primerStart = 0; primerEnd = 0; return false;   }       //can't tell the difference between multiple primers
+                       else{  return true; }
+               }
+       
+        primerStart = 0; primerEnd = 0;
+               return false;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimOligos", "stripForward");
+               exit(1);
+       }
+}
+//******************************************************************/
+bool TrimOligos::findReverse(Sequence& seq, int& primerStart, int& primerEnd){
+       try {
+               
+               string rawSequence = seq.getUnaligned();
+               
+               for(int i=0;i<revPrimer.size();i++){
+                       string oligo = revPrimer[i];
+                       if(rawSequence.length() < oligo.length()) {  break;  }
+                       
+                       //search for primer
+            int olength = oligo.length();
+            for (int j = rawSequence.length()-olength; j >= 0; j--){
+                if (m->control_pressed) {  primerStart = 0; primerEnd = 0; return false; }
+                string rawChunk = rawSequence.substr(j, olength);
+                
+                if(compareDNASeq(oligo, rawChunk)) {
+                    primerStart = j;
+                    primerEnd = primerStart + olength;
+                    return true;
+                }
+                
+            }
+               }       
+               
+        primerStart = 0; primerEnd = 0;
+               return false;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "PcrSeqsCommand", "findReverse");
+               exit(1);
+       }
+}
 //*******************************************************************/
+
 int TrimOligos::stripBarcode(Sequence& seq, QualityScores& qual, int& group){
        try {
                
@@ -259,7 +405,7 @@ int TrimOligos::stripBarcode(Sequence& seq, QualityScores& qual, int& group){
        }
 }
 //*******************************************************************/
-int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, QualityScores& forwardQual, QualityScores& reverseQual, int& group){
+int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, int& group){
        try {
                //look for forward barcode
                string rawFSequence = forwardSeq.getUnaligned();
@@ -284,8 +430,6 @@ int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                                group = it->first;
                                forwardSeq.setUnaligned(rawFSequence.substr(foligo.length()));
                 reverseSeq.setUnaligned(rawRSequence.substr(0,(rawRSequence.length()-roligo.length())));
-                forwardQual.trimQScores(foligo.length(), -1);
-                reverseQual.trimQScores(-1, rawRSequence.length()-roligo.length());
                                success = 0;
                                break;
                        }
@@ -468,8 +612,6 @@ int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                     if (foundMatch) {
                         forwardSeq.setUnaligned(rawFSequence.substr(fStart));
                         reverseSeq.setUnaligned(rawRSequence.substr(rStart));
-                        forwardQual.trimQScores(fStart, -1);
-                        reverseQual.trimQScores(rStart, -1);
                         success = minDiff;
                     }else { success = bdiffs + 100;    }
                 }
@@ -488,24 +630,24 @@ int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, Quality
        
 }
 //*******************************************************************/
-int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, QualityScores& forwardQual, QualityScores& reverseQual, int& group){
+int TrimOligos::stripBarcode(Sequence& forwardSeq, Sequence& reverseSeq, QualityScores& forwardQual, QualityScores& reverseQual, int& group){
        try {
                //look for forward barcode
                string rawFSequence = forwardSeq.getUnaligned();
         string rawRSequence = reverseSeq.getUnaligned();
-               int success = pdiffs + 1;       //guilty until proven innocent
+               int success = bdiffs + 1;       //guilty until proven innocent
                
                //can you find the forward barcode
-               for(map<int,oligosPair>::iterator it=ipprimers.begin();it!=ipprimers.end();it++){
+               for(map<int,oligosPair>::iterator it=ipbarcodes.begin();it!=ipbarcodes.end();it++){
                        string foligo = it->second.forward;
             string roligo = it->second.reverse;
             
                        if(rawFSequence.length() < foligo.length()){    //let's just assume that the barcodes are the same length
-                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               success = bdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
                                break;  
                        }
             if(rawRSequence.length() < roligo.length()){       //let's just assume that the barcodes are the same length
-                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               success = bdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
                                break;  
                        }
                        
@@ -521,10 +663,10 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                }
                
                //if you found the barcode or if you don't want to allow for diffs
-               if ((pdiffs == 0) || (success == 0)) { return success;  }
+               if ((bdiffs == 0) || (success == 0)) { return success;  }
                else { //try aligning and see if you can find it
                        Alignment* alignment;
-                       if (ifprimers.size() > 0) {  alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxFPrimerLength+pdiffs+1));   }
+                       if (ifbarcodes.size() > 0) {  alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxFBarcodeLength+bdiffs+1));   }
                        else{ alignment = NULL; } 
                        
                        //can you find the barcode
@@ -549,16 +691,16 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
              but if best match forward = 4, and reverse = 1, we want to count as a valid match because forward 1 and forward 4 are the same.  so both barcodes map to same group.
              */
             //cout << endl << forwardSeq.getName() << endl;         
-                       for(map<string, vector<int> >::iterator it=ifprimers.begin();it!=ifprimers.end();it++){
+                       for(map<string, vector<int> >::iterator it=ifbarcodes.begin();it!=ifbarcodes.end();it++){
                                string oligo = it->first;
                                
-                               if(rawFSequence.length() < maxFPrimerLength){   //let's just assume that the barcodes are the same length
-                                       success = pdiffs + 10;
+                               if(rawFSequence.length() < maxFBarcodeLength){  //let's just assume that the barcodes are the same length
+                                       success = bdiffs + 10;
                                        break;
                                }
-                               //cout << "before = " << oligo << '\t' << rawFSequence.substr(0,oligo.length()+pdiffs) << endl;
+                               //cout << "before = " << oligo << '\t' << rawFSequence.substr(0,oligo.length()+bdiffs) << endl;
                                //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
-                               alignment->align(oligo, rawFSequence.substr(0,oligo.length()+pdiffs));
+                               alignment->align(oligo, rawFSequence.substr(0,oligo.length()+bdiffs));
                                oligo = alignment->getSeqAAln();
                                string temp = alignment->getSeqBAln();
                 
@@ -569,7 +711,7 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                                temp = temp.substr(0,alnLength);
                 int numDiff = countDiffs(oligo, temp);
                                
-                if (alnLength == 0) { numDiff = pdiffs + 100; }
+                if (alnLength == 0) { numDiff = bdiffs + 100; }
                 //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
                 
                                if(numDiff < minDiff){
@@ -598,12 +740,12 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                        }
             
                        //cout << minDiff << '\t' << minCount << '\t' << endl;
-                       if(minDiff > pdiffs)    {       success = minDiff;              }       //no good matches
+                       if(minDiff > bdiffs)    {       success = minDiff;              }       //no good matches
                        else{   
                 //check for reverse match
                 if (alignment != NULL) {  delete alignment;  }
                 
-                if (irbarcodes.size() > 0) { alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxRPrimerLength+pdiffs+1));   }
+                if (irbarcodes.size() > 0) { alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxRBarcodeLength+bdiffs+1));   }
                 else{ alignment = NULL; } 
                 
                 //can you find the barcode
@@ -612,16 +754,16 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                 vector< vector<int> > minRGroup;
                 vector<int> minRPos;
                 
-                for(map<string, vector<int> >::iterator it=irprimers.begin();it!=irprimers.end();it++){
+                for(map<string, vector<int> >::iterator it=irbarcodes.begin();it!=irbarcodes.end();it++){
                     string oligo = it->first;
-                    //cout << "before = " << oligo << '\t' << rawRSequence.substr(0,oligo.length()+pdiffs) << endl;
-                    if(rawRSequence.length() < maxRPrimerLength){      //let's just assume that the barcodes are the same length
-                        success = pdiffs + 10;
+                    //cout << "before = " << oligo << '\t' << rawRSequence.substr(0,oligo.length()+bdiffs) << endl;
+                    if(rawRSequence.length() < maxRBarcodeLength){     //let's just assume that the barcodes are the same length
+                        success = bdiffs + 10;
                         break;
                     }
                     
                     //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
-                    alignment->align(oligo, rawRSequence.substr(0,oligo.length()+pdiffs));
+                    alignment->align(oligo, rawRSequence.substr(0,oligo.length()+bdiffs));
                     oligo = alignment->getSeqAAln();
                     string temp = alignment->getSeqBAln();
                     
@@ -630,7 +772,7 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                     oligo = oligo.substr(0,alnLength);
                     temp = temp.substr(0,alnLength);
                     int numDiff = countDiffs(oligo, temp);
-                    if (alnLength == 0) { numDiff = pdiffs + 100; }
+                    if (alnLength == 0) { numDiff = bdiffs + 100; }
                     
                     //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
                     if(numDiff < minDiff){
@@ -672,7 +814,7 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                  cout << endl;
                  }
                  cout << endl;*/
-                if(minDiff > pdiffs)   {       success = minDiff;              }       //no good matches
+                if(minDiff > bdiffs)   {       success = minDiff;              }       //no good matches
                 else  {  
                     bool foundMatch = false;
                     vector<int> matches;
@@ -700,6 +842,460 @@ int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, Quality
                         forwardQual.trimQScores(fStart, -1);
                         reverseQual.trimQScores(rStart, -1);
                         success = minDiff;
+                    }else { success = bdiffs + 100;    }
+                }
+                       }
+                       
+                       if (alignment != NULL) {  delete alignment;  }
+               }
+               
+               return success;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimOligos", "stripIBarcode");
+               exit(1);
+       }
+       
+}
+//*******************************************************************/
+int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, QualityScores& forwardQual, QualityScores& reverseQual, int& group){
+       try {
+               //look for forward barcode
+               string rawFSequence = forwardSeq.getUnaligned();
+        string rawRSequence = reverseSeq.getUnaligned();
+               int success = pdiffs + 1;       //guilty until proven innocent
+               
+               //can you find the forward barcode
+               for(map<int,oligosPair>::iterator it=ipprimers.begin();it!=ipprimers.end();it++){
+                       string foligo = it->second.forward;
+            string roligo = it->second.reverse;
+            
+                       if(rawFSequence.length() < foligo.length()){    //let's just assume that the barcodes are the same length
+                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               break;  
+                       }
+            if(rawRSequence.length() < roligo.length()){       //let's just assume that the barcodes are the same length
+                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               break;  
+                       }
+                       
+                       if((compareDNASeq(foligo, rawFSequence.substr(0,foligo.length()))) && (compareDNASeq(roligo, rawRSequence.substr((rawRSequence.length()-roligo.length()),roligo.length())))) {
+                               group = it->first;
+                               forwardSeq.setUnaligned(rawFSequence.substr(foligo.length()));
+                reverseSeq.setUnaligned(rawRSequence.substr(0,(rawRSequence.length()-roligo.length())));
+                forwardQual.trimQScores(foligo.length(), -1);
+                reverseQual.trimQScores(-1, rawRSequence.length()-roligo.length());
+                               success = 0;
+                               break;
+                       }
+               }
+               
+               //if you found the barcode or if you don't want to allow for diffs
+               if ((pdiffs == 0) || (success == 0)) { return success;  }
+               else { //try aligning and see if you can find it
+                       Alignment* alignment;
+                       if (ifprimers.size() > 0) {  alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxFPrimerLength+pdiffs+1));   }
+                       else{ alignment = NULL; } 
+                       
+                       //can you find the barcode
+                       int minDiff = 1e6;
+                       int minCount = 1;
+                       vector< vector<int> > minFGroup;
+                       vector<int> minFPos; 
+            
+            //the pair can have duplicates, but we only want to search for the unique forward and reverses, example
+            /*
+             1   Sarah   Westcott
+             2   John    Westcott
+             3   Anna    Westcott
+             4   Sarah   Schloss
+             5   Pat     Schloss
+             6   Gail    Brown
+             7   Pat     Moore
+             
+             only want to look for forward = Sarah, John, Anna, Pat, Gail
+             reverse = Westcott, Schloss, Brown, Moore
+             
+             but if best match forward = 4, and reverse = 1, we want to count as a valid match because forward 1 and forward 4 are the same.  so both barcodes map to same group.
+             */
+            //cout << endl << forwardSeq.getName() << endl;         
+                       for(map<string, vector<int> >::iterator it=ifprimers.begin();it!=ifprimers.end();it++){
+                               string oligo = it->first;
+                               
+                               if(rawFSequence.length() < maxFPrimerLength){   //let's just assume that the barcodes are the same length
+                                       success = pdiffs + 10;
+                                       break;
+                               }
+                               //cout << "before = " << oligo << '\t' << rawFSequence.substr(0,oligo.length()+pdiffs) << endl;
+                               //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
+                               alignment->align(oligo, rawFSequence.substr(0,oligo.length()+pdiffs));
+                               oligo = alignment->getSeqAAln();
+                               string temp = alignment->getSeqBAln();
+                
+                               int alnLength = oligo.length();
+                               
+                               for(int i=oligo.length()-1;i>=0;i--){ if(oligo[i] != '-'){      alnLength = i+1;        break;  } }
+                               oligo = oligo.substr(0,alnLength);
+                               temp = temp.substr(0,alnLength);
+                int numDiff = countDiffs(oligo, temp);
+                               
+                if (alnLength == 0) { numDiff = pdiffs + 100; }
+                //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
+                
+                               if(numDiff < minDiff){
+                                       minDiff = numDiff;
+                                       minCount = 1;
+                                       minFGroup.clear();
+                    minFGroup.push_back(it->second);
+                                       int tempminFPos = 0;
+                    minFPos.clear();
+                                       for(int i=0;i<alnLength;i++){
+                                               if(temp[i] != '-'){
+                                                       tempminFPos++;
+                                               }
+                                       }
+                    minFPos.push_back(tempminFPos);
+                               }else if(numDiff == minDiff){
+                                       minFGroup.push_back(it->second);
+                    int tempminFPos = 0;
+                                       for(int i=0;i<alnLength;i++){
+                                               if(temp[i] != '-'){
+                                                       tempminFPos++;
+                                               }
+                                       }
+                    minFPos.push_back(tempminFPos);
+                               }
+                       }
+            
+                       //cout << minDiff << '\t' << minCount << '\t' << endl;
+                       if(minDiff > pdiffs)    {       success = minDiff;              }       //no good matches
+                       else{   
+                //check for reverse match
+                if (alignment != NULL) {  delete alignment;  }
+                
+                if (irbarcodes.size() > 0) { alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxRPrimerLength+pdiffs+1));   }
+                else{ alignment = NULL; } 
+                
+                //can you find the barcode
+                minDiff = 1e6;
+                minCount = 1;
+                vector< vector<int> > minRGroup;
+                vector<int> minRPos;
+                
+                for(map<string, vector<int> >::iterator it=irprimers.begin();it!=irprimers.end();it++){
+                    string oligo = it->first;
+                    //cout << "before = " << oligo << '\t' << rawRSequence.substr(0,oligo.length()+pdiffs) << endl;
+                    if(rawRSequence.length() < maxRPrimerLength){      //let's just assume that the barcodes are the same length
+                        success = pdiffs + 10;
+                        break;
+                    }
+                    
+                    //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
+                    alignment->align(oligo, rawRSequence.substr(0,oligo.length()+pdiffs));
+                    oligo = alignment->getSeqAAln();
+                    string temp = alignment->getSeqBAln();
+                    
+                    int alnLength = oligo.length();
+                    for(int i=oligo.length()-1;i>=0;i--){ if(oligo[i] != '-'){ alnLength = i+1;        break;  } }
+                    oligo = oligo.substr(0,alnLength);
+                    temp = temp.substr(0,alnLength);
+                    int numDiff = countDiffs(oligo, temp);
+                    if (alnLength == 0) { numDiff = pdiffs + 100; }
+                    
+                    //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
+                    if(numDiff < minDiff){
+                        minDiff = numDiff;
+                        minCount = 1;
+                        minRGroup.clear();
+                        minRGroup.push_back(it->second);
+                        int tempminRPos = 0;
+                        minRPos.clear();
+                        for(int i=0;i<alnLength;i++){
+                            if(temp[i] != '-'){
+                                tempminRPos++;
+                            }
+                        }
+                        minRPos.push_back(tempminRPos);                    
+                    }else if(numDiff == minDiff){
+                        int tempminRPos = 0;
+                        for(int i=0;i<alnLength;i++){
+                            if(temp[i] != '-'){
+                                tempminRPos++;
+                            }
+                        }
+                        minRPos.push_back(tempminRPos);  
+                        minRGroup.push_back(it->second);
+                    }
+                    
+                }
+                
+                /*cout << minDiff << '\t' << minCount << '\t' << endl;
+                 for (int i = 0; i < minFGroup.size(); i++) { 
+                 cout << i << '\t';
+                 for (int j = 0; j < minFGroup[i].size(); j++) {  cout << minFGroup[i][j] << " "; }
+                 cout << endl;
+                 }
+                 cout << endl;
+                 for (int i = 0; i < minRGroup.size(); i++) { 
+                 cout << i << '\t';
+                 for (int j = 0; j < minRGroup[i].size(); j++) {  cout << minRGroup[i][j] << " "; }
+                 cout << endl;
+                 }
+                 cout << endl;*/
+                if(minDiff > pdiffs)   {       success = minDiff;              }       //no good matches
+                else  {  
+                    bool foundMatch = false;
+                    vector<int> matches;
+                    for (int i = 0; i < minFGroup.size(); i++) {
+                        for (int j = 0; j < minFGroup[i].size(); j++) {
+                            for (int k = 0; k < minRGroup.size(); k++) {
+                                if (m->inUsersGroups(minFGroup[i][j], minRGroup[k])) { matches.push_back(minFGroup[i][j]); k+= minRGroup.size(); }
+                            }
+                        }
+                    }
+                    
+                    int fStart = 0;
+                    int rStart = 0;
+                    if (matches.size() == 1) { 
+                        foundMatch = true;
+                        group = matches[0]; 
+                        fStart = minFPos[0];
+                        rStart = minRPos[0];
+                    }
+                    
+                    //we have an acceptable match for the forward and reverse, but do they match?
+                    if (foundMatch) {
+                        forwardSeq.setUnaligned(rawFSequence.substr(fStart));
+                        reverseSeq.setUnaligned(rawRSequence.substr(rStart));
+                        forwardQual.trimQScores(fStart, -1);
+                        reverseQual.trimQScores(rStart, -1);
+                        success = minDiff;
+                    }else { success = pdiffs + 100;    }
+                }
+                       }
+                       
+                       if (alignment != NULL) {  delete alignment;  }
+               }
+               
+               return success;
+               
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimOligos", "stripIForward");
+               exit(1);
+       }
+       
+}
+//*******************************************************************/
+int TrimOligos::stripForward(Sequence& forwardSeq, Sequence& reverseSeq, int& group){
+       try {
+               //look for forward barcode
+               string rawFSequence = forwardSeq.getUnaligned();
+        string rawRSequence = reverseSeq.getUnaligned();
+               int success = pdiffs + 1;       //guilty until proven innocent
+               
+               //can you find the forward barcode
+               for(map<int,oligosPair>::iterator it=ipprimers.begin();it!=ipprimers.end();it++){
+                       string foligo = it->second.forward;
+            string roligo = it->second.reverse;
+            
+                       if(rawFSequence.length() < foligo.length()){    //let's just assume that the barcodes are the same length
+                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               break;  
+                       }
+            if(rawRSequence.length() < roligo.length()){       //let's just assume that the barcodes are the same length
+                               success = pdiffs + 10;                                  //if the sequence is shorter than the barcode then bail out
+                               break;  
+                       }
+                       
+                       if((compareDNASeq(foligo, rawFSequence.substr(0,foligo.length()))) && (compareDNASeq(roligo, rawRSequence.substr((rawRSequence.length()-roligo.length()),roligo.length())))) {
+                               group = it->first;
+                               forwardSeq.setUnaligned(rawFSequence.substr(foligo.length()));
+                reverseSeq.setUnaligned(rawRSequence.substr(0,(rawRSequence.length()-roligo.length())));
+                               success = 0;
+                               break;
+                       }
+               }
+               
+               //if you found the barcode or if you don't want to allow for diffs
+               if ((pdiffs == 0) || (success == 0)) { return success;  }
+               else { //try aligning and see if you can find it
+                       Alignment* alignment;
+                       if (ifprimers.size() > 0) {  alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxFPrimerLength+pdiffs+1));   }
+                       else{ alignment = NULL; } 
+                       
+                       //can you find the barcode
+                       int minDiff = 1e6;
+                       int minCount = 1;
+                       vector< vector<int> > minFGroup;
+                       vector<int> minFPos; 
+            
+            //the pair can have duplicates, but we only want to search for the unique forward and reverses, example
+            /*
+             1   Sarah   Westcott
+             2   John    Westcott
+             3   Anna    Westcott
+             4   Sarah   Schloss
+             5   Pat     Schloss
+             6   Gail    Brown
+             7   Pat     Moore
+             
+             only want to look for forward = Sarah, John, Anna, Pat, Gail
+             reverse = Westcott, Schloss, Brown, Moore
+             
+             but if best match forward = 4, and reverse = 1, we want to count as a valid match because forward 1 and forward 4 are the same.  so both barcodes map to same group.
+             */
+            //cout << endl << forwardSeq.getName() << endl;         
+                       for(map<string, vector<int> >::iterator it=ifprimers.begin();it!=ifprimers.end();it++){
+                               string oligo = it->first;
+                               
+                               if(rawFSequence.length() < maxFPrimerLength){   //let's just assume that the barcodes are the same length
+                                       success = pdiffs + 10;
+                                       break;
+                               }
+                               //cout << "before = " << oligo << '\t' << rawFSequence.substr(0,oligo.length()+pdiffs) << endl;
+                               //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
+                               alignment->align(oligo, rawFSequence.substr(0,oligo.length()+pdiffs));
+                               oligo = alignment->getSeqAAln();
+                               string temp = alignment->getSeqBAln();
+                
+                               int alnLength = oligo.length();
+                               
+                               for(int i=oligo.length()-1;i>=0;i--){ if(oligo[i] != '-'){      alnLength = i+1;        break;  } }
+                               oligo = oligo.substr(0,alnLength);
+                               temp = temp.substr(0,alnLength);
+                int numDiff = countDiffs(oligo, temp);
+                               
+                if (alnLength == 0) { numDiff = pdiffs + 100; }
+                //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
+                
+                               if(numDiff < minDiff){
+                                       minDiff = numDiff;
+                                       minCount = 1;
+                                       minFGroup.clear();
+                    minFGroup.push_back(it->second);
+                                       int tempminFPos = 0;
+                    minFPos.clear();
+                                       for(int i=0;i<alnLength;i++){
+                                               if(temp[i] != '-'){
+                                                       tempminFPos++;
+                                               }
+                                       }
+                    minFPos.push_back(tempminFPos);
+                               }else if(numDiff == minDiff){
+                                       minFGroup.push_back(it->second);
+                    int tempminFPos = 0;
+                                       for(int i=0;i<alnLength;i++){
+                                               if(temp[i] != '-'){
+                                                       tempminFPos++;
+                                               }
+                                       }
+                    minFPos.push_back(tempminFPos);
+                               }
+                       }
+            
+                       //cout << minDiff << '\t' << minCount << '\t' << endl;
+                       if(minDiff > pdiffs)    {       success = minDiff;              }       //no good matches
+                       else{   
+                //check for reverse match
+                if (alignment != NULL) {  delete alignment;  }
+                
+                if (irbarcodes.size() > 0) { alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxRPrimerLength+pdiffs+1));   }
+                else{ alignment = NULL; } 
+                
+                //can you find the barcode
+                minDiff = 1e6;
+                minCount = 1;
+                vector< vector<int> > minRGroup;
+                vector<int> minRPos;
+                
+                for(map<string, vector<int> >::iterator it=irprimers.begin();it!=irprimers.end();it++){
+                    string oligo = it->first;
+                    //cout << "before = " << oligo << '\t' << rawRSequence.substr(0,oligo.length()+pdiffs) << endl;
+                    if(rawRSequence.length() < maxRPrimerLength){      //let's just assume that the barcodes are the same length
+                        success = pdiffs + 10;
+                        break;
+                    }
+                    
+                    //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
+                    alignment->align(oligo, rawRSequence.substr(0,oligo.length()+pdiffs));
+                    oligo = alignment->getSeqAAln();
+                    string temp = alignment->getSeqBAln();
+                    
+                    int alnLength = oligo.length();
+                    for(int i=oligo.length()-1;i>=0;i--){ if(oligo[i] != '-'){ alnLength = i+1;        break;  } }
+                    oligo = oligo.substr(0,alnLength);
+                    temp = temp.substr(0,alnLength);
+                    int numDiff = countDiffs(oligo, temp);
+                    if (alnLength == 0) { numDiff = pdiffs + 100; }
+                    
+                    //cout << "after = " << oligo << '\t' << temp << '\t' << numDiff << endl;
+                    if(numDiff < minDiff){
+                        minDiff = numDiff;
+                        minCount = 1;
+                        minRGroup.clear();
+                        minRGroup.push_back(it->second);
+                        int tempminRPos = 0;
+                        minRPos.clear();
+                        for(int i=0;i<alnLength;i++){
+                            if(temp[i] != '-'){
+                                tempminRPos++;
+                            }
+                        }
+                        minRPos.push_back(tempminRPos);                    
+                    }else if(numDiff == minDiff){
+                        int tempminRPos = 0;
+                        for(int i=0;i<alnLength;i++){
+                            if(temp[i] != '-'){
+                                tempminRPos++;
+                            }
+                        }
+                        minRPos.push_back(tempminRPos);  
+                        minRGroup.push_back(it->second);
+                    }
+                    
+                }
+                
+                /*cout << minDiff << '\t' << minCount << '\t' << endl;
+                 for (int i = 0; i < minFGroup.size(); i++) { 
+                 cout << i << '\t';
+                 for (int j = 0; j < minFGroup[i].size(); j++) {  cout << minFGroup[i][j] << " "; }
+                 cout << endl;
+                 }
+                 cout << endl;
+                 for (int i = 0; i < minRGroup.size(); i++) { 
+                 cout << i << '\t';
+                 for (int j = 0; j < minRGroup[i].size(); j++) {  cout << minRGroup[i][j] << " "; }
+                 cout << endl;
+                 }
+                 cout << endl;*/
+                if(minDiff > pdiffs)   {       success = minDiff;              }       //no good matches
+                else  {  
+                    bool foundMatch = false;
+                    vector<int> matches;
+                    for (int i = 0; i < minFGroup.size(); i++) {
+                        for (int j = 0; j < minFGroup[i].size(); j++) {
+                            for (int k = 0; k < minRGroup.size(); k++) {
+                                if (m->inUsersGroups(minFGroup[i][j], minRGroup[k])) { matches.push_back(minFGroup[i][j]); k+= minRGroup.size(); }
+                            }
+                        }
+                    }
+                    
+                    int fStart = 0;
+                    int rStart = 0;
+                    if (matches.size() == 1) { 
+                        foundMatch = true;
+                        group = matches[0]; 
+                        fStart = minFPos[0];
+                        rStart = minRPos[0];
+                    }
+                    
+                    //we have an acceptable match for the forward and reverse, but do they match?
+                    if (foundMatch) {
+                        forwardSeq.setUnaligned(rawFSequence.substr(fStart));
+                        reverseSeq.setUnaligned(rawRSequence.substr(rStart));
+                        success = minDiff;
                     }else { success = pdiffs + 100;    }
                 }
                        }