]> git.donarmstrong.com Git - mothur.git/blobdiff - trimseqscommand.cpp
added modify names parameter to set.dir
[mothur.git] / trimseqscommand.cpp
index ed2b04642694362ec4af9247f4f8814d90c54ebb..16d83de2596b9cf84ab4069789917878fa89559e 100644 (file)
@@ -21,6 +21,7 @@ vector<string> TrimSeqsCommand::setParameters(){
                CommandParameter pname("name", "InputTypes", "", "", "namecount", "none", "none","name",false,false,true); parameters.push_back(pname);
         CommandParameter pcount("count", "InputTypes", "", "", "namecount", "none", "none","count",false,false,true); parameters.push_back(pcount);
                CommandParameter pflip("flip", "Boolean", "", "F", "", "", "","",false,false,true); parameters.push_back(pflip);
+        CommandParameter preorient("checkorient", "Boolean", "", "F", "", "", "","",false,false,true); parameters.push_back(preorient);
                CommandParameter pmaxambig("maxambig", "Number", "", "-1", "", "", "","",false,false); parameters.push_back(pmaxambig);
                CommandParameter pmaxhomop("maxhomop", "Number", "", "0", "", "", "","",false,false); parameters.push_back(pmaxhomop);
                CommandParameter pminlength("minlength", "Number", "", "0", "", "", "","",false,false); parameters.push_back(pminlength);
@@ -60,9 +61,10 @@ string TrimSeqsCommand::getHelpString(){
                string helpString = "";
                helpString += "The trim.seqs command reads a fastaFile and creates 2 new fasta files, .trim.fasta and scrap.fasta, as well as group files if you provide and oligos file.\n";
                helpString += "The .trim.fasta contains sequences that meet your requirements, and the .scrap.fasta contains those which don't.\n";
-               helpString += "The trim.seqs command parameters are fasta, name, count, flip, oligos, maxambig, maxhomop, minlength, maxlength, qfile, qthreshold, qaverage, diffs, qtrim, keepfirst, removelast and allfiles.\n";
+               helpString += "The trim.seqs command parameters are fasta, name, count, flip, checkorient, oligos, maxambig, maxhomop, minlength, maxlength, qfile, qthreshold, qaverage, diffs, qtrim, keepfirst, removelast and allfiles.\n";
                helpString += "The fasta parameter is required.\n";
                helpString += "The flip parameter will output the reverse compliment of your trimmed sequence. The default is false.\n";
+        helpString += "The checkorient parameter will check the reverse compliment of the sequence if the barcodes and primers cannot be found in the forward. The default is false.\n";
                helpString += "The oligos parameter allows you to provide an oligos file.\n";
                helpString += "The name parameter allows you to provide a names file with your fasta file.\n";
         helpString += "The count parameter allows you to provide a count file with your fasta file.\n";
@@ -123,7 +125,7 @@ string TrimSeqsCommand::getOutputPattern(string type) {
 
 TrimSeqsCommand::TrimSeqsCommand(){    
        try {
-               abort = true; calledHelp = true; 
+               abort = true; calledHelp = true;
                setParameters();
                vector<string> tempOutNames;
                outputTypes["fasta"] = tempOutNames;
@@ -326,6 +328,9 @@ TrimSeqsCommand::TrimSeqsCommand(string option)  {
             
             temp = validParameter.validFile(parameters, "keepforward", false);         if (temp == "not found") { temp = "F"; }
                        keepforward = m->isTrue(temp);
+            
+            temp = validParameter.validFile(parameters, "checkorient", false);         if (temp == "not found") { temp = "F"; }
+                       reorient = m->isTrue(temp);
                        
                        temp = validParameter.validFile(parameters, "processors", false);       if (temp == "not found"){       temp = m->getProcessors();      }
                        m->setProcessors(temp);
@@ -366,6 +371,7 @@ int TrimSeqsCommand::execute(){
        
                if (abort == true) { if (calledHelp) { return 0; }  return 2;   }
                
+        pairedOligos = false;
                numFPrimers = 0;  //this needs to be initialized
                numRPrimers = 0;
         numSpacers = 0;
@@ -416,7 +422,7 @@ int TrimSeqsCommand::execute(){
                
                if (countfile != "") {
             CountTable ct;
-            ct.readTable(countfile);
+            ct.readTable(countfile, true);
             nameCount = ct.getNameMap();
                        outputNames.push_back(trimCountFile);
                        outputNames.push_back(scrapCountFile);
@@ -437,7 +443,11 @@ int TrimSeqsCommand::execute(){
                                outputNames.push_back(outputGroupFileName); outputTypes["group"].push_back(outputGroupFileName);
                        }
                }
-       
+        
+        if (!pairedOligos) { if (reorient) { m->mothurOut("[WARNING]: You cannot use reorient without paired barcodes or primers, skipping."); m->mothurOutEndLine(); reorient = false; } }
+        
+        if (m->control_pressed) { return 0; }
+            
         //fills lines and qlines
                setLines(fastaFile, qFileName);
                
@@ -530,7 +540,7 @@ int TrimSeqsCommand::execute(){
             
             if (countfile != "") { //create countfile with group info included
                 CountTable* ct = new CountTable();
-                ct->readTable(trimCountFile);
+                ct->readTable(trimCountFile, true);
                 map<string, int> justTrimmedNames = ct->getNameMap();
                 delete ct;
                 
@@ -666,11 +676,32 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
                
                int count = 0;
                bool moreSeqs = 1;
-               TrimOligos trimOligos(pdiffs, bdiffs, ldiffs, sdiffs, primers, barcodes, revPrimer, linker, spacer);
-       
+        int numBarcodes = barcodes.size();
+               TrimOligos* trimOligos = NULL;
+        if (pairedOligos)   {   trimOligos = new TrimOligos(pdiffs, bdiffs, 0, 0, pairedPrimers, pairedBarcodes); numBarcodes = pairedBarcodes.size(); }
+        else                {   trimOligos = new TrimOligos(pdiffs, bdiffs, ldiffs, sdiffs, primers, barcodes, revPrimer, linker, spacer);  }
+        
+        TrimOligos* rtrimOligos = NULL;
+        if (reorient) {
+            //create reoriented primer and barcode pairs
+            map<int, oligosPair> rpairedPrimers, rpairedBarcodes;
+            for (map<int, oligosPair>::iterator it = pairedPrimers.begin(); it != pairedPrimers.end(); it++) {
+                  oligosPair tempPair(reverseOligo((it->second).reverse), (reverseOligo((it->second).forward))); //reversePrimer, rc ForwardPrimer
+                rpairedPrimers[it->first] = tempPair;
+                //cout  << reverseOligo((it->second).reverse) << '\t' << (reverseOligo((it->second).forward)) << '\t' << primerNameVector[it->first] << endl;
+            }
+            for (map<int, oligosPair>::iterator it = pairedBarcodes.begin(); it != pairedBarcodes.end(); it++) {
+                 oligosPair tempPair(reverseOligo((it->second).reverse), (reverseOligo((it->second).forward))); //reverseBarcode, rc ForwardBarcode
+                rpairedBarcodes[it->first] = tempPair;
+                 //cout  << reverseOligo((it->second).reverse) << '\t' << (reverseOligo((it->second).forward)) << '\t' << barcodeNameVector[it->first] << endl;
+            }
+            rtrimOligos = new TrimOligos(pdiffs, bdiffs, 0, 0, rpairedPrimers, rpairedBarcodes); numBarcodes = rpairedBarcodes.size();
+        }
+        
                while (moreSeqs) {
                                
-                       if (m->control_pressed) { 
+                       if (m->control_pressed) {
+                delete trimOligos; if (reorient) { delete rtrimOligos; }
                                inFASTA.close(); trimFASTAFile.close(); scrapFASTAFile.close();
                                if ((createGroup) && (countfile == "")) {        outGroupsFile.close();   }
                 if(qFileName != "")    {       qFile.close();  scrapQualFile.close(); trimQualFile.close();    }
@@ -685,13 +716,15 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
 
                        Sequence currSeq(inFASTA); m->gobble(inFASTA);
                        //cout << currSeq.getName() << '\t' << currSeq.getUnaligned().length() << endl;
+            Sequence savedSeq(currSeq.getName(), currSeq.getAligned());
             
-                       QualityScores currQual;
+                       QualityScores currQual; QualityScores savedQual;
                        if(qFileName != ""){
                                currQual = QualityScores(qFile);  m->gobble(qFile);
+                savedQual.setName(currQual.getName()); savedQual.setScores(currQual.getScores());
                 //cout << currQual.getName() << endl;
                        }
-                       
+                         
                        string origSeq = currSeq.getUnaligned();
                        if (origSeq != "") {
                                
@@ -699,38 +732,79 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
                                int primerIndex = 0;
                                
                 if(numLinkers != 0){
-                                       success = trimOligos.stripLinker(currSeq, currQual);
+                                       success = trimOligos->stripLinker(currSeq, currQual);
                                        if(success > ldiffs)            {       trashCode += 'k';       }
                                        else{ currentSeqsDiffs += success;  }
 
                                }
                 
-                               if(barcodes.size() != 0){
-                                       success = trimOligos.stripBarcode(currSeq, currQual, barcodeIndex);
-                                       if(success > bdiffs)            {       trashCode += 'b';       }
+                               if(numBarcodes != 0){
+                                       success = trimOligos->stripBarcode(currSeq, currQual, barcodeIndex);
+                                       if(success > bdiffs)            {
+                        trashCode += 'b';
+                    }
                                        else{ currentSeqsDiffs += success;  }
                                }
                                
                 if(numSpacers != 0){
-                                       success = trimOligos.stripSpacer(currSeq, currQual);
+                                       success = trimOligos->stripSpacer(currSeq, currQual);
                                        if(success > sdiffs)            {       trashCode += 's';       }
                                        else{ currentSeqsDiffs += success;  }
 
                                }
                 
                                if(numFPrimers != 0){
-                                       success = trimOligos.stripForward(currSeq, currQual, primerIndex, keepforward);
-                                       if(success > pdiffs)            {       trashCode += 'f';       }
+                                       success = trimOligos->stripForward(currSeq, currQual, primerIndex, keepforward);
+                                       if(success > pdiffs)            {
+                          trashCode += 'f';  
+                    }
                                        else{ currentSeqsDiffs += success;  }
                                }
                                
                                if (currentSeqsDiffs > tdiffs)  {       trashCode += 't';   }
                                
                                if(numRPrimers != 0){
-                                       success = trimOligos.stripReverse(currSeq, currQual);
+                                       success = trimOligos->stripReverse(currSeq, currQual);
                                        if(!success)                            {       trashCode += 'r';       }
                                }
-
+                
+                if (reorient && (trashCode != "")) { //if you failed and want to check the reverse
+                    int thisSuccess = 0;
+                    string thisTrashCode = "";
+                    int thisCurrentSeqsDiffs = 0;
+                    
+                    int thisBarcodeIndex = 0;
+                    int thisPrimerIndex = 0;
+                    
+                    if(numBarcodes != 0){
+                        thisSuccess = rtrimOligos->stripBarcode(savedSeq, savedQual, thisBarcodeIndex);
+                        if(thisSuccess > bdiffs)               { thisTrashCode += "b"; }
+                        else{ thisCurrentSeqsDiffs += thisSuccess;  }
+                    }
+                    
+                    if(numFPrimers != 0){
+                        thisSuccess = rtrimOligos->stripForward(savedSeq, savedQual, thisPrimerIndex, keepforward);
+                        if(thisSuccess > pdiffs)               { thisTrashCode += "f"; }
+                        else{ thisCurrentSeqsDiffs += thisSuccess;  }
+                    }
+                   
+                    if (thisCurrentSeqsDiffs > tdiffs) {       thisTrashCode += 't';   }
+                    
+                    if (thisTrashCode == "") { 
+                        trashCode = thisTrashCode;
+                        success = thisSuccess;
+                        currentSeqsDiffs = thisCurrentSeqsDiffs;
+                        barcodeIndex = thisBarcodeIndex;
+                        primerIndex = thisPrimerIndex;
+                        savedSeq.reverseComplement();
+                        currSeq.setAligned(savedSeq.getAligned());
+                        if(qFileName != ""){
+                            savedQual.flipQScores();
+                            currQual.setScores(savedQual.getScores());
+                        }
+                    }else { trashCode += "(" + thisTrashCode + ")";  }
+                }
+                
                                if(keepFirst != 0){
                                        success = keepFirstTrim(currSeq, currQual);
                                }
@@ -779,33 +853,11 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
                 if (m->debug) { m->mothurOut("[DEBUG]: " + currSeq.getName() + ", trashcode= " + trashCode); if (trashCode.length() != 0) { m->mothurOutEndLine(); } }
                 
                                if(trashCode.length() == 0){
-                                       currSeq.setAligned(currSeq.getUnaligned());
-                                       currSeq.printSequence(trimFASTAFile);
-                                       
-                                       if(qFileName != ""){
-                                               currQual.printQScores(trimQualFile);
-                                       }
-                                       
-                    
-                                       if(nameFile != ""){
-                                               map<string, string>::iterator itName = nameMap.find(currSeq.getName());
-                                               if (itName != nameMap.end()) {  trimNameFile << itName->first << '\t' << itName->second << endl; }
-                                               else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }
-                                       }
-                    
-                    int numRedundants = 0;
-                    if (countfile != "") {
-                        map<string, int>::iterator itCount = nameCount.find(currSeq.getName());
-                        if (itCount != nameCount.end()) { 
-                            trimCountFile << itCount->first << '\t' << itCount->second << endl;
-                            numRedundants = itCount->second-1;
-                        }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your count file, please correct."); m->mothurOutEndLine(); }
-                    }
-                                       
-                                       if (createGroup) {
-                                               if(barcodes.size() != 0){
-                                                       string thisGroup = barcodeNameVector[barcodeIndex];
-                                                       if (primers.size() != 0) { 
+                    string thisGroup = "";
+                    if (createGroup) {
+                                               if(numBarcodes != 0){
+                                                       thisGroup = barcodeNameVector[barcodeIndex];
+                                                       if (numFPrimers != 0) {
                                                                if (primerNameVector[primerIndex] != "") { 
                                                                        if(thisGroup != "") {
                                                                                thisGroup += "." + primerNameVector[primerIndex]; 
@@ -814,52 +866,83 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
                                                                        }
                                                                } 
                                                        }
-                                                       
-                            if (m->debug) { m->mothurOut(", group= " + thisGroup + "\n"); }
-                            
-                                                       if (countfile == "") { outGroupsFile << currSeq.getName() << '\t' << thisGroup << endl; }
-                            else {   groupMap[currSeq.getName()] = thisGroup; }
-                                                       
-                                                       if (nameFile != "") {
-                                                               map<string, string>::iterator itName = nameMap.find(currSeq.getName());
-                                                               if (itName != nameMap.end()) { 
-                                                                       vector<string> thisSeqsNames; 
-                                                                       m->splitAtChar(itName->second, thisSeqsNames, ',');
-                                    numRedundants = thisSeqsNames.size()-1; //we already include ourselves below
-                                                                       for (int k = 1; k < thisSeqsNames.size(); k++) { //start at 1 to skip self
-                                                                               outGroupsFile << thisSeqsNames[k] << '\t' << thisGroup << endl;
-                                                                       }
-                                                               }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }                                                   
-                                                       }
-                                                       
-                                                       map<string, int>::iterator it = groupCounts.find(thisGroup);
-                                                       if (it == groupCounts.end()) {  groupCounts[thisGroup] = 1 + numRedundants; }
-                                                       else { groupCounts[it->first] += (1 + numRedundants); }
+                        }
+                    }
+                    
+                    int pos = thisGroup.find("ignore");
+                    if (pos == string::npos) {
+                        currSeq.setAligned(currSeq.getUnaligned());
+                        currSeq.printSequence(trimFASTAFile);
+                        
+                        if(qFileName != ""){
+                            currQual.printQScores(trimQualFile);
+                        }
+                        
+                        
+                        if(nameFile != ""){
+                            map<string, string>::iterator itName = nameMap.find(currSeq.getName());
+                            if (itName != nameMap.end()) {  trimNameFile << itName->first << '\t' << itName->second << endl; }
+                            else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }
+                        }
+                        
+                        int numRedundants = 0;
+                        if (countfile != "") {
+                            map<string, int>::iterator itCount = nameCount.find(currSeq.getName());
+                            if (itCount != nameCount.end()) { 
+                                trimCountFile << itCount->first << '\t' << itCount->second << endl;
+                                numRedundants = itCount->second-1;
+                            }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your count file, please correct."); m->mothurOutEndLine(); }
+                        }
+                        
+                        if (createGroup) {
+                            if(numBarcodes != 0){
+                                                                
+                                if (m->debug) { m->mothurOut(", group= " + thisGroup + "\n"); }
+                                
+                                if (countfile == "") { outGroupsFile << currSeq.getName() << '\t' << thisGroup << endl; }
+                                else {   groupMap[currSeq.getName()] = thisGroup; }
+                                
+                                if (nameFile != "") {
+                                    map<string, string>::iterator itName = nameMap.find(currSeq.getName());
+                                    if (itName != nameMap.end()) { 
+                                        vector<string> thisSeqsNames; 
+                                        m->splitAtChar(itName->second, thisSeqsNames, ',');
+                                        numRedundants = thisSeqsNames.size()-1; //we already include ourselves below
+                                        for (int k = 1; k < thisSeqsNames.size(); k++) { //start at 1 to skip self
+                                            outGroupsFile << thisSeqsNames[k] << '\t' << thisGroup << endl;
+                                        }
+                                    }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }                                                      
+                                }
+                                
+                                map<string, int>::iterator it = groupCounts.find(thisGroup);
+                                if (it == groupCounts.end()) { groupCounts[thisGroup] = 1 + numRedundants; }
+                                else { groupCounts[it->first] += (1 + numRedundants); }
                                                                
-                                               }
-                                       }
-                                       
-                                       if(allFiles){
-                                               ofstream output;
-                                               m->openOutputFileAppend(fastaFileNames[barcodeIndex][primerIndex], output);
-                                               currSeq.printSequence(output);
-                                               output.close();
-                                               
-                                               if(qFileName != ""){
-                                                       m->openOutputFileAppend(qualFileNames[barcodeIndex][primerIndex], output);
-                                                       currQual.printQScores(output);
-                                                       output.close();                                                 
-                                               }
-                                               
-                                               if(nameFile != ""){
-                                                       map<string, string>::iterator itName = nameMap.find(currSeq.getName());
-                                                       if (itName != nameMap.end()) { 
-                                                               m->openOutputFileAppend(nameFileNames[barcodeIndex][primerIndex], output);
-                                                               output << itName->first << '\t' << itName->second << endl; 
-                                                               output.close();
-                                                       }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }
-                                               }
-                                       }
+                            }
+                        }
+                        
+                        if(allFiles){
+                            ofstream output;
+                            m->openOutputFileAppend(fastaFileNames[barcodeIndex][primerIndex], output);
+                            currSeq.printSequence(output);
+                            output.close();
+                            
+                            if(qFileName != ""){
+                                m->openOutputFileAppend(qualFileNames[barcodeIndex][primerIndex], output);
+                                currQual.printQScores(output);
+                                output.close();                                                        
+                            }
+                            
+                            if(nameFile != ""){
+                                map<string, string>::iterator itName = nameMap.find(currSeq.getName());
+                                if (itName != nameMap.end()) { 
+                                    m->openOutputFileAppend(nameFileNames[barcodeIndex][primerIndex], output);
+                                    output << itName->first << '\t' << itName->second << endl; 
+                                    output.close();
+                                }else { m->mothurOut("[ERROR]: " + currSeq.getName() + " is not in your namefile, please correct."); m->mothurOutEndLine(); }
+                            }
+                        }
+                    }
                                }
                                else{
                                        if(nameFile != ""){ //needs to be before the currSeq name is changed
@@ -900,7 +983,8 @@ int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string
                //report progress
                if((count) % 1000 != 0){        m->mothurOut(toString(count)); m->mothurOutEndLine();           }
                
-               
+               delete trimOligos;
+        if (reorient) { delete rtrimOligos; }
                inFASTA.close();
                trimFASTAFile.close();
                scrapFASTAFile.close();
@@ -1088,10 +1172,10 @@ int TrimSeqsCommand::createProcessesCreateTrim(string filename, string qFileName
                                               tempPrimerQualFileNames,
                                               tempNameFileNames,
                                               lines[h].start, lines[h].end, qLines[h].start, qLines[h].end, m,
-                                              pdiffs, bdiffs, ldiffs, sdiffs, tdiffs, primers, barcodes, revPrimer, linker, spacer, 
+                                              pdiffs, bdiffs, ldiffs, sdiffs, tdiffs, primers, barcodes, revPrimer, linker, spacer, pairedBarcodes, pairedPrimers, pairedOligos,
                                              primerNameVector, barcodeNameVector, createGroup, allFiles, keepforward, keepFirst, removeLast,
                                               qWindowStep, qWindowSize, qWindowAverage, qtrim, qThreshold, qAverage, qRollAverage,
-                                             minLength, maxAmbig, maxHomoP, maxLength, flip, nameMap, nameCount);
+                                             minLength, maxAmbig, maxHomoP, maxLength, flip, reorient, nameMap, nameCount);
                        pDataArray.push_back(tempTrim);
             
                        hThreadArray[h] = CreateThread(NULL, 0, MyTrimThreadFunction, pDataArray[h], 0, &dwThreadIdArray[h]);   
@@ -1143,6 +1227,9 @@ int TrimSeqsCommand::createProcessesCreateTrim(string filename, string qFileName
                
                //Close all thread handles and free memory allocations.
                for(int i=0; i < pDataArray.size(); i++){
+            if (pDataArray[i]->count != pDataArray[i]->lineEnd) {
+                m->mothurOut("[ERROR]: process " + toString(i) + " only processed " + toString(pDataArray[i]->count) + " of " + toString(pDataArray[i]->lineEnd) + " sequences assigned to it, quitting. \n"); m->control_pressed = true;
+            }
                        for (map<string, int>::iterator it = pDataArray[i]->groupCounts.begin(); it != pDataArray[i]->groupCounts.end(); it++) {
                 map<string, int>::iterator it2 = groupCounts.find(it->first);
                 if (it2 == groupCounts.end()) {        groupCounts[it->first] = it->second; }
@@ -1304,6 +1391,8 @@ int TrimSeqsCommand::setLines(string filename, string qfilename) {
                         string sname = "";  nameStream >> sname;
                         sname = sname.substr(1);
                         
+                        m->checkName(sname);
+                        
                         map<string, int>::iterator it = firstSeqNames.find(sname);
                         
                         if(it != firstSeqNames.end()) { //this is the start of a new chunk
@@ -1402,10 +1491,15 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                
                ofstream test;
                
-               string type, oligo, group;
+               string type, oligo, roligo, group;
+        bool hasPrimer = false; bool hasPairedBarcodes = false;
 
                int indexPrimer = 0;
                int indexBarcode = 0;
+        int indexPairedPrimer = 0;
+               int indexPairedBarcode = 0;
+        set<string> uniquePrimers;
+        set<string> uniqueBarcodes;
                
                while(!inOligos.eof()){
 
@@ -1437,7 +1531,7 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                                        // get rest of line in case there is a primer name
                                        while (!inOligos.eof()) {       
                                                char c = inOligos.get(); 
-                                               if (c == 10 || c == 13){        break;  }
+                                               if (c == 10 || c == 13 || c == -1){     break;  }
                                                else if (c == 32 || c == 9){;} //space or tab
                                                else {  group += c;  }
                                        } 
@@ -1451,6 +1545,42 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                                        primers[oligo]=indexPrimer; indexPrimer++;              
                                        primerNameVector.push_back(group);
                                }
+                else if (type == "PRIMER"){
+                    m->gobble(inOligos);
+                                       
+                    inOligos >> roligo;
+                    
+                    for(int i=0;i<roligo.length();i++){
+                        roligo[i] = toupper(roligo[i]);
+                        if(roligo[i] == 'U')   {       roligo[i] = 'T';        }
+                    }
+                    roligo = reverseOligo(roligo);
+                    
+                    group = "";
+                    
+                                       // get rest of line in case there is a primer name
+                                       while (!inOligos.eof()) {
+                                               char c = inOligos.get();
+                                               if (c == 10 || c == 13 || c == -1){     break;  }
+                                               else if (c == 32 || c == 9){;} //space or tab
+                                               else {  group += c;  }
+                                       }
+                    
+                    oligosPair newPrimer(oligo, roligo);
+                    
+                     if (m->debug) { m->mothurOut("[DEBUG]: primer pair " + newPrimer.forward + " " + newPrimer.reverse + ", and group = " + group + ".\n"); }
+                                       
+                                       //check for repeat barcodes
+                    string tempPair = oligo+roligo;
+                    if (uniquePrimers.count(tempPair) != 0) { m->mothurOut("primer pair " + newPrimer.forward + " " + newPrimer.reverse + " is in your oligos file already."); m->mothurOutEndLine();  }
+                    else { uniquePrimers.insert(tempPair); }
+                                       
+                    if (m->debug) {  if (group != "") { m->mothurOut("[DEBUG]: reading group " + group + ".\n"); }else{ m->mothurOut("[DEBUG]: no group for primer pair " + newPrimer.forward + " " + newPrimer.reverse + ".\n"); }  }
+                    
+                                       pairedPrimers[indexPairedPrimer]=newPrimer; indexPairedPrimer++;
+                                       primerNameVector.push_back(group);
+                    hasPrimer = true;
+                }
                                else if(type == "REVERSE"){
                                        //Sequence oligoRC("reverse", oligo);
                                        //oligoRC.reverseComplement();
@@ -1459,13 +1589,49 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                                }
                                else if(type == "BARCODE"){
                                        inOligos >> group;
-                                       
-                                       //check for repeat barcodes
-                                       map<string, int>::iterator itBar = barcodes.find(oligo);
-                                       if (itBar != barcodes.end()) { m->mothurOut("barcode " + oligo + " is in your oligos file already."); m->mothurOutEndLine();  }
                     
-                                       barcodes[oligo]=indexBarcode; indexBarcode++;
-                                       barcodeNameVector.push_back(group);
+                    //barcode lines can look like   BARCODE   atgcatgc   groupName  - for 454 seqs
+                    //or                            BARCODE   atgcatgc   atgcatgc    groupName  - for illumina data that has forward and reverse info
+                    
+                    string temp = "";
+                    while (!inOligos.eof())    {
+                                               char c = inOligos.get();
+                                               if (c == 10 || c == 13 || c == -1){     break;  }
+                                               else if (c == 32 || c == 9){;} //space or tab
+                                               else {  temp += c;  }
+                                       }
+                                       
+                    //then this is illumina data with 4 columns
+                    if (temp != "") {
+                        hasPairedBarcodes = true;
+                        string reverseBarcode = group; //reverseOligo(group); //reverse barcode
+                        group = temp;
+                        
+                        for(int i=0;i<reverseBarcode.length();i++){
+                            reverseBarcode[i] = toupper(reverseBarcode[i]);
+                            if(reverseBarcode[i] == 'U')       {       reverseBarcode[i] = 'T';        }
+                        }
+                        
+                        reverseBarcode = reverseOligo(reverseBarcode);
+                        oligosPair newPair(oligo, reverseBarcode);
+                        
+                        if (m->debug) { m->mothurOut("[DEBUG]: barcode pair " + newPair.forward + " " + newPair.reverse + ", and group = " + group + ".\n"); }
+                        
+                        //check for repeat barcodes
+                        string tempPair = oligo+reverseBarcode;
+                        if (uniqueBarcodes.count(tempPair) != 0) { m->mothurOut("barcode pair " + newPair.forward + " " + newPair.reverse +  " is in your oligos file already, disregarding."); m->mothurOutEndLine();  }
+                        else { uniqueBarcodes.insert(tempPair); }
+                        
+                        pairedBarcodes[indexPairedBarcode]=newPair; indexPairedBarcode++;
+                        barcodeNameVector.push_back(group);
+                    }else {                            
+                        //check for repeat barcodes
+                        map<string, int>::iterator itBar = barcodes.find(oligo);
+                        if (itBar != barcodes.end()) { m->mothurOut("barcode " + oligo + " is in your oligos file already."); m->mothurOutEndLine();  }
+                        
+                        barcodes[oligo]=indexBarcode; indexBarcode++;
+                        barcodeNameVector.push_back(group);
+                    }
                                }else if(type == "LINKER"){
                                        linker.push_back(oligo);
                                }else if(type == "SPACER"){
@@ -1477,8 +1643,13 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                }       
                inOligos.close();
                
+        if (hasPairedBarcodes || hasPrimer) {
+            pairedOligos = true;
+            if ((primers.size() != 0) || (barcodes.size() != 0) || (linker.size() != 0) || (spacer.size() != 0) || (revPrimer.size() != 0)) { m->control_pressed = true;  m->mothurOut("[ERROR]: cannot mix paired primers and barcodes with non paired or linkers and spacers, quitting."); m->mothurOutEndLine();  return 0; }
+        }else if (reorient) { m->mothurOut("[Warning]: cannot use checkorient without paired barcodes or primers, ignoring.\n"); m->mothurOutEndLine(); reorient = false; }
+        
                if(barcodeNameVector.size() == 0 && primerNameVector[0] == ""){ allFiles = 0;   }
-               
+        
                //add in potential combos
                if(barcodeNameVector.size() == 0){
                        barcodes[""] = 0;
@@ -1499,72 +1670,146 @@ bool TrimSeqsCommand::getOligos(vector<vector<string> >& fastaFileNames, vector<
                
                if(allFiles){
                        set<string> uniqueNames; //used to cleanup outputFileNames
-                       for(map<string, int>::iterator itBar = barcodes.begin();itBar != barcodes.end();itBar++){
-                               for(map<string, int>::iterator itPrimer = primers.begin();itPrimer != primers.end(); itPrimer++){
-                                       
-                                       string primerName = primerNameVector[itPrimer->second];
-                                       string barcodeName = barcodeNameVector[itBar->second];
-                                       
-                                       string comboGroupName = "";
-                                       string fastaFileName = "";
-                                       string qualFileName = "";
-                                       string nameFileName = "";
-                    string countFileName = "";
-                                       
-                                       if(primerName == ""){
-                                               comboGroupName = barcodeNameVector[itBar->second];
-                                       }
-                                       else{
-                                               if(barcodeName == ""){
-                                                       comboGroupName = primerNameVector[itPrimer->second];
-                                               }
-                                               else{
-                                                       comboGroupName = barcodeNameVector[itBar->second] + "." + primerNameVector[itPrimer->second];
-                                               }
-                                       }
-                                       
-                                       
-                                       ofstream temp;
-                    map<string, string> variables; 
-                    variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(fastaFile));
-                    variables["[tag]"] = comboGroupName;
-                                       fastaFileName = getOutputFileName("fasta", variables);
-                                       if (uniqueNames.count(fastaFileName) == 0) {
-                                               outputNames.push_back(fastaFileName);
-                                               outputTypes["fasta"].push_back(fastaFileName);
-                                               uniqueNames.insert(fastaFileName);
-                                       }
-                                       
-                                       fastaFileNames[itBar->second][itPrimer->second] = fastaFileName;
-                                       m->openOutputFile(fastaFileName, temp);         temp.close();
-                                       
-                                       if(qFileName != ""){
-                        variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(qFileName));
-                                               qualFileName = getOutputFileName("qfile", variables);
-                                               if (uniqueNames.count(qualFileName) == 0) {
-                                                       outputNames.push_back(qualFileName);
-                                                       outputTypes["qfile"].push_back(qualFileName);
-                                               }
-                                               
-                                               qualFileNames[itBar->second][itPrimer->second] = qualFileName;
-                                               m->openOutputFile(qualFileName, temp);          temp.close();
-                                       }
-                                       
-                                       if(nameFile != ""){
-                        variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(nameFile));
-                                               nameFileName = getOutputFileName("name", variables);
-                                               if (uniqueNames.count(nameFileName) == 0) {
-                                                       outputNames.push_back(nameFileName);
-                                                       outputTypes["name"].push_back(nameFileName);
-                                               }
-                                               
-                                               nameFileNames[itBar->second][itPrimer->second] = nameFileName;
-                                               m->openOutputFile(nameFileName, temp);          temp.close();
-                                       }
-                               }
-                       }
+            if (pairedOligos) {
+                for(map<int, oligosPair>::iterator itBar = pairedBarcodes.begin();itBar != pairedBarcodes.end();itBar++){
+                    for(map<int, oligosPair>::iterator itPrimer = pairedPrimers.begin();itPrimer != pairedPrimers.end(); itPrimer++){
+                        
+                        string primerName = primerNameVector[itPrimer->first];
+                        string barcodeName = barcodeNameVector[itBar->first];
+                        
+                        if ((primerName == "ignore") || (barcodeName == "ignore")) { } //do nothing
+                        else {
+                            string comboGroupName = "";
+                            string fastaFileName = "";
+                            string qualFileName = "";
+                            string nameFileName = "";
+                            string countFileName = "";
+                            
+                            if(primerName == ""){
+                                comboGroupName = barcodeNameVector[itBar->first];
+                            }
+                            else{
+                                if(barcodeName == ""){
+                                    comboGroupName = primerNameVector[itPrimer->first];
+                                }
+                                else{
+                                    comboGroupName = barcodeNameVector[itBar->first] + "." + primerNameVector[itPrimer->first];
+                                }
+                            }
+                            
+                            
+                            ofstream temp;
+                            map<string, string> variables;
+                            variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(fastaFile));
+                            variables["[tag]"] = comboGroupName;
+                            fastaFileName = getOutputFileName("fasta", variables);
+                            if (uniqueNames.count(fastaFileName) == 0) {
+                                outputNames.push_back(fastaFileName);
+                                outputTypes["fasta"].push_back(fastaFileName);
+                                uniqueNames.insert(fastaFileName);
+                            }
+                            
+                            fastaFileNames[itBar->first][itPrimer->first] = fastaFileName;
+                            m->openOutputFile(fastaFileName, temp);            temp.close();
+                            
+                            if(qFileName != ""){
+                                variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(qFileName));
+                                qualFileName = getOutputFileName("qfile", variables);
+                                if (uniqueNames.count(qualFileName) == 0) {
+                                    outputNames.push_back(qualFileName);
+                                    outputTypes["qfile"].push_back(qualFileName);
+                                }
+                                
+                                qualFileNames[itBar->first][itPrimer->first] = qualFileName;
+                                m->openOutputFile(qualFileName, temp);         temp.close();
+                            }
+                            
+                            if(nameFile != ""){
+                                variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(nameFile));
+                                nameFileName = getOutputFileName("name", variables);
+                                if (uniqueNames.count(nameFileName) == 0) {
+                                    outputNames.push_back(nameFileName);
+                                    outputTypes["name"].push_back(nameFileName);
+                                }
+                                
+                                nameFileNames[itBar->first][itPrimer->first] = nameFileName;
+                                m->openOutputFile(nameFileName, temp);         temp.close();
+                            }
+                        }
+                    }
+                }
+            }else {
+                for(map<string, int>::iterator itBar = barcodes.begin();itBar != barcodes.end();itBar++){
+                    for(map<string, int>::iterator itPrimer = primers.begin();itPrimer != primers.end(); itPrimer++){
+                        
+                        string primerName = primerNameVector[itPrimer->second];
+                        string barcodeName = barcodeNameVector[itBar->second];
+                        
+                        if ((primerName == "ignore") || (barcodeName == "ignore")) { } //do nothing 
+                        else {
+                            string comboGroupName = "";
+                            string fastaFileName = "";
+                            string qualFileName = "";
+                            string nameFileName = "";
+                            string countFileName = "";
+                            
+                            if(primerName == ""){
+                                comboGroupName = barcodeNameVector[itBar->second];
+                            }
+                            else{
+                                if(barcodeName == ""){
+                                    comboGroupName = primerNameVector[itPrimer->second];
+                                }
+                                else{
+                                    comboGroupName = barcodeNameVector[itBar->second] + "." + primerNameVector[itPrimer->second];
+                                }
+                            }
+                            
+                            
+                            ofstream temp;
+                            map<string, string> variables; 
+                            variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(fastaFile));
+                            variables["[tag]"] = comboGroupName;
+                            fastaFileName = getOutputFileName("fasta", variables);
+                            if (uniqueNames.count(fastaFileName) == 0) {
+                                outputNames.push_back(fastaFileName);
+                                outputTypes["fasta"].push_back(fastaFileName);
+                                uniqueNames.insert(fastaFileName);
+                            }
+                            
+                            fastaFileNames[itBar->second][itPrimer->second] = fastaFileName;
+                            m->openOutputFile(fastaFileName, temp);            temp.close();
+                            
+                            if(qFileName != ""){
+                                variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(qFileName));
+                                qualFileName = getOutputFileName("qfile", variables);
+                                if (uniqueNames.count(qualFileName) == 0) {
+                                    outputNames.push_back(qualFileName);
+                                    outputTypes["qfile"].push_back(qualFileName);
+                                }
+                                
+                                qualFileNames[itBar->second][itPrimer->second] = qualFileName;
+                                m->openOutputFile(qualFileName, temp);         temp.close();
+                            }
+                            
+                            if(nameFile != ""){
+                                variables["[filename]"] = outputDir + m->getRootName(m->getSimpleName(nameFile));
+                                nameFileName = getOutputFileName("name", variables);
+                                if (uniqueNames.count(nameFileName) == 0) {
+                                    outputNames.push_back(nameFileName);
+                                    outputTypes["name"].push_back(nameFileName);
+                                }
+                                
+                                nameFileNames[itBar->second][itPrimer->second] = nameFileName;
+                                m->openOutputFile(nameFileName, temp);         temp.close();
+                            }
+                        }
+                    }
+                }
+            }
                }
                numFPrimers = primers.size();
+        if (pairedOligos) { numFPrimers  = pairedPrimers.size(); }
                numRPrimers = revPrimer.size();
         numLinkers = linker.size();
         numSpacers = spacer.size();
@@ -1605,7 +1850,13 @@ bool TrimSeqsCommand::keepFirstTrim(Sequence& sequence, QualityScores& qscores){
                if(qscores.getName() != ""){
                        qscores.trimQScores(-1, keepFirst);
                }
+
+//        sequence.printSequence(cout);cout << endl;
+        
                sequence.trim(keepFirst);
+        
+//        sequence.printSequence(cout);cout << endl << endl;;
+
                return success;
        }
        catch(exception& e) {