]> git.donarmstrong.com Git - mothur.git/blobdiff - trimseqscommand.cpp
modified trim.seqs to account for primers of different lengths
[mothur.git] / trimseqscommand.cpp
index 1b5c7cf1416f9a8d4fe68ac3a1c6d7e751eb7d18..b12da1cfe126bbdfc562172772093cf84e733457 100644 (file)
@@ -8,6 +8,7 @@
  */
 
 #include "trimseqscommand.h"
+#include "needlemanoverlap.hpp"
 
 //***************************************************************************************************************
 
@@ -22,7 +23,7 @@ TrimSeqsCommand::TrimSeqsCommand(string option)  {
                else {
                        //valid paramters for this command
                        string AlignArray[] =  {"fasta", "flip", "oligos", "maxambig", "maxhomop", "minlength", "maxlength", "qfile", 
-                                                                       "qthreshold", "qaverage", "allfiles", "qtrim", "outputdir","inputdir"};
+                                                                       "qthreshold", "qaverage", "allfiles", "qtrim","tdiffs", "pdiffs", "bdiffs", "processors", "outputdir","inputdir"};
                        
                        vector<string> myArray (AlignArray, AlignArray+(sizeof(AlignArray)/sizeof(string)));
                        
@@ -103,6 +104,15 @@ TrimSeqsCommand::TrimSeqsCommand(string option)  {
                        temp = validParameter.validFile(parameters, "maxlength", false);        if (temp == "not found") { temp = "0"; }
                        convert(temp, maxLength);
                        
+                       temp = validParameter.validFile(parameters, "tdiffs", false);           if (temp == "not found") { temp = "0"; }
+                       convert(temp, tdiffs);
+                       
+                       temp = validParameter.validFile(parameters, "bdiffs", false);           if (temp == "not found") { temp = "0"; }
+                       convert(temp, bdiffs);
+                       
+                       temp = validParameter.validFile(parameters, "pdiffs", false);           if (temp == "not found") { temp = "0"; }
+                       convert(temp, pdiffs);
+                       
                        temp = validParameter.validFile(parameters, "qfile", true);     
                        if (temp == "not found")        {       qFileName = "";         }
                        else if(temp == "not open")     {       abort = true;           }
@@ -120,6 +130,9 @@ TrimSeqsCommand::TrimSeqsCommand(string option)  {
                        temp = validParameter.validFile(parameters, "allfiles", false);         if (temp == "not found") { temp = "F"; }
                        allFiles = isTrue(temp);
                        
+                       temp = validParameter.validFile(parameters, "processors", false);       if (temp == "not found"){       temp = "1";                             }
+                       convert(temp, processors); 
+                       
                        if(allFiles && oligoFile == ""){
                                m->mothurOut("You selected allfiles, but didn't enter an oligos file.  Ignoring the allfiles request."); m->mothurOutEndLine();
                        }
@@ -145,7 +158,7 @@ TrimSeqsCommand::TrimSeqsCommand(string option)  {
 void TrimSeqsCommand::help(){
        try {
                m->mothurOut("The trim.seqs command reads a fastaFile and creates .....\n");
-               m->mothurOut("The trim.seqs command parameters are fasta, flip, oligos, maxambig, maxhomop, minlength, maxlength, qfile, qthreshold, qaverage, qtrim and allfiles.\n");
+               m->mothurOut("The trim.seqs command parameters are fasta, flip, oligos, maxambig, maxhomop, minlength, maxlength, qfile, qthreshold, qaverage, diffs, qtrim and allfiles.\n");
                m->mothurOut("The fasta parameter is required.\n");
                m->mothurOut("The flip parameter .... The default is 0.\n");
                m->mothurOut("The oligos parameter .... The default is "".\n");
@@ -153,6 +166,9 @@ void TrimSeqsCommand::help(){
                m->mothurOut("The maxhomop parameter .... The default is 0.\n");
                m->mothurOut("The minlength parameter .... The default is 0.\n");
                m->mothurOut("The maxlength parameter .... The default is 0.\n");
+               m->mothurOut("The tdiffs parameter is used to specify the total number of differences allowed in the sequence. The default is 0.\n");
+               m->mothurOut("The bdiffs parameter is used to specify the number of differences allowed in the barcode. The default is 0.\n");
+               m->mothurOut("The pdiffs parameter is used to specify the number of differences allowed in the primer. The default is 0.\n");
                m->mothurOut("The qfile parameter .....\n");
                m->mothurOut("The qthreshold parameter .... The default is 0.\n");
                m->mothurOut("The qaverage parameter .... The default is 0.\n");
@@ -184,45 +200,172 @@ int TrimSeqsCommand::execute(){
        
                if (abort == true) { return 0; }
                
-               vector<string> outputNames;
-               
                numFPrimers = 0;  //this needs to be initialized
                numRPrimers = 0;
                
-               ifstream inFASTA;
-               openInputFile(fastaFile, inFASTA);
-               
-               ofstream outFASTA;
                string trimSeqFile = outputDir + getRootName(getSimpleName(fastaFile)) + "trim.fasta";
-               openOutputFile(trimSeqFile, outFASTA);
                outputNames.push_back(trimSeqFile);
+               string scrapSeqFile = outputDir + getRootName(getSimpleName(fastaFile)) + "scrap.fasta";
+               outputNames.push_back(scrapSeqFile);
+               string groupFile = outputDir + getRootName(getSimpleName(fastaFile)) + "groups";
                
-               ofstream outGroups;
-               vector<ofstream*> fastaFileNames;
+               vector<string> fastaFileNames;
                if(oligoFile != ""){
-                       string groupFile = outputDir + getRootName(getSimpleName(fastaFile)) + "groups"; 
-                       openOutputFile(groupFile, outGroups);
                        outputNames.push_back(groupFile);
                        getOligos(fastaFileNames);
                }
                
+               if(qFileName != "")     {       setLines(qFileName, qLines);    }
+
+               
+               #if defined (__APPLE__) || (__MACH__) || (linux) || (__linux)
+                               if(processors == 1){
+                                       ifstream inFASTA;
+                                       openInputFile(fastaFile, inFASTA);
+                                       int numSeqs=count(istreambuf_iterator<char>(inFASTA),istreambuf_iterator<char>(), '>');
+                                       inFASTA.close();
+                                       
+                                       lines.push_back(new linePair(0, numSeqs));
+                                       
+                                       driverCreateTrim(fastaFile, qFileName, trimSeqFile, scrapSeqFile, groupFile, fastaFileNames, lines[0], lines[0]);
+                                       
+                                       for (int j = 0; j < fastaFileNames.size(); j++) {
+                                               rename((fastaFileNames[j] + toString(getpid()) + ".temp").c_str(), fastaFileNames[j].c_str());
+                                       }
+
+                               }else{
+                                       setLines(fastaFile, lines);     
+                                       if(qFileName == "")     {       qLines = lines; }       
+                                                               
+                                       createProcessesCreateTrim(fastaFile, qFileName, trimSeqFile, scrapSeqFile, groupFile, fastaFileNames); 
+                                       
+                                       rename((trimSeqFile + toString(processIDS[0]) + ".temp").c_str(), trimSeqFile.c_str());
+                                       rename((scrapSeqFile + toString(processIDS[0]) + ".temp").c_str(), scrapSeqFile.c_str());
+                                       rename((groupFile + toString(processIDS[0]) + ".temp").c_str(), groupFile.c_str());
+                                       for (int j = 0; j < fastaFileNames.size(); j++) {
+                                               rename((fastaFileNames[j] + toString(processIDS[0]) + ".temp").c_str(), fastaFileNames[j].c_str());
+                                       }
+                                       //append files
+                                       for(int i=1;i<processors;i++){
+                                               appendFiles((trimSeqFile + toString(processIDS[i]) + ".temp"), trimSeqFile);
+                                               remove((trimSeqFile + toString(processIDS[i]) + ".temp").c_str());
+                                               appendFiles((scrapSeqFile + toString(processIDS[i]) + ".temp"), scrapSeqFile);
+                                               remove((scrapSeqFile + toString(processIDS[i]) + ".temp").c_str());
+                                               appendFiles((groupFile + toString(processIDS[i]) + ".temp"), groupFile);
+                                               remove((groupFile + toString(processIDS[i]) + ".temp").c_str());
+                                               for (int j = 0; j < fastaFileNames.size(); j++) {
+                                                       appendFiles((fastaFileNames[j] + toString(processIDS[i]) + ".temp"), fastaFileNames[j]);
+                                                       remove((fastaFileNames[j] + toString(processIDS[i]) + ".temp").c_str());
+                                               }
+                                       }
+                               }
+                               
+                               if (m->control_pressed) {  return 0; }
+               #else
+                               ifstream inFASTA;
+                               openInputFile(fastafileNames[s], inFASTA);
+                               numSeqs=count(istreambuf_iterator<char>(inFASTA),istreambuf_iterator<char>(), '>');
+                               inFASTA.close();
+                               
+                               lines.push_back(new linePair(0, numSeqs));
+                               
+                               driverCreateSummary(fastafile, qFileName, trimSeqFile, scrapSeqFile, groupFile, fastaFileNames, lines[0], lines[0]);
+                               
+                               if (m->control_pressed) {  return 0; }
+               #endif
+                                               
+                                                                               
+               for(int i=0;i<fastaFileNames.size();i++){
+                       ifstream inFASTA;
+                       string seqName;
+                       openInputFile(getRootName(fastaFile) + groupVector[i] + ".fasta", inFASTA);
+                       ofstream outGroups;
+                       openOutputFile(outputDir + getRootName(getSimpleName(fastaFile)) + groupVector[i] + ".groups", outGroups);
+                       outputNames.push_back(outputDir + getRootName(getSimpleName(fastaFile)) + groupVector[i] + ".groups");
+                       
+                       while(!inFASTA.eof()){
+                               if(inFASTA.get() == '>'){
+                                       inFASTA >> seqName;
+                                       outGroups << seqName << '\t' << groupVector[i] << endl;
+                               }
+                               while (!inFASTA.eof())  {       char c = inFASTA.get(); if (c == 10 || c == 13){        break;  }       }
+                       }
+                       outGroups.close();
+                       inFASTA.close();
+               }
+               
+               if (m->control_pressed) { 
+                       for (int i = 0; i < outputNames.size(); i++) {  remove(outputNames[i].c_str()); }
+                       return 0;
+               }
+
+               m->mothurOutEndLine();
+               m->mothurOut("Output File Names: "); m->mothurOutEndLine();
+               for (int i = 0; i < outputNames.size(); i++) {  m->mothurOut(outputNames[i]); m->mothurOutEndLine();    }
+               m->mothurOutEndLine();
+               
+               return 0;       
+                       
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimSeqsCommand", "execute");
+               exit(1);
+       }
+}
+               
+/**************************************************************************************/
+int TrimSeqsCommand::driverCreateTrim(string filename, string qFileName, string trimFile, string scrapFile, string groupFile, vector<string> fastaNames, linePair* line, linePair* qline) {    
+       try {
+               
+               ofstream outFASTA;
+               int able = openOutputFile(trimFile, outFASTA);
+               
                ofstream scrapFASTA;
-               string scrapSeqFile = outputDir + getRootName(getSimpleName(fastaFile)) + "scrap.fasta";
-               openOutputFile(scrapSeqFile, scrapFASTA);
-               outputNames.push_back(scrapSeqFile);
+               openOutputFile(scrapFile, scrapFASTA);
+               
+               ofstream outGroups;
+               vector<ofstream*> fastaFileNames;
+               if (oligoFile != "") {          
+                       openOutputFile(groupFile, outGroups);   
+                       for (int i = 0; i < fastaNames.size(); i++) {
+                               fastaFileNames.push_back(new ofstream((fastaNames[i] + toString(getpid()) + ".temp").c_str(), ios::ate)); 
+                       }
+               }
+               
+               ifstream inFASTA;
+               openInputFile(filename, inFASTA);
                
                ifstream qFile;
                if(qFileName != "")     {       openInputFile(qFileName, qFile);        }
                
-               bool success;
-
-               while(!inFASTA.eof()){
+               qFile.seekg(qline->start);
+               inFASTA.seekg(line->start);
+               
+               for(int i=0;i<line->num;i++){
+                               
+                       if (m->control_pressed) { 
+                               inFASTA.close(); 
+                               outFASTA.close();
+                               scrapFASTA.close();
+                               if (oligoFile != "") {   outGroups.close();   }
+                               if(qFileName != "")     {       qFile.close();  }
+                               for(int i=0;i<fastaFileNames.size();i++){
+                                       fastaFileNames[i]->close();
+                                       delete fastaFileNames[i];
+                               }       
+                               for (int i = 0; i < outputNames.size(); i++) {  remove(outputNames[i].c_str()); }
+                               return 0;
+                       }
+                       
+                       bool success = 1;
+                       
                        Sequence currSeq(inFASTA);
 
                        string origSeq = currSeq.getUnaligned();
                        if (origSeq != "") {
                                int group;
                                string trashCode = "";
+                               int currentSeqsDiffs = 0;
                                
                                if(qFileName != ""){
                                        if(qThreshold != 0)             {       success = stripQualThreshold(currSeq, qFile);   }
@@ -236,13 +379,17 @@ int TrimSeqsCommand::execute(){
                                if(barcodes.size() != 0){
                                        success = stripBarcode(currSeq, group);
                                        if(!success){   trashCode += 'b';       }
+                                       else{ currentSeqsDiffs += currentSeqsTdiffs;  }
                                }
                        
                                if(numFPrimers != 0){
                                        success = stripForward(currSeq);
                                        if(!success){   trashCode += 'f';       }
+                                       else{ currentSeqsDiffs += currentSeqsTdiffs;  }
                                }
-                                       
+                               
+                               if (currentSeqsDiffs > tdiffs) { trashCode += 't';   }
+
                                if(numRPrimers != 0){
                                        success = stripReverse(currSeq);
                                        if(!success){   trashCode += 'r';       }
@@ -282,10 +429,11 @@ int TrimSeqsCommand::execute(){
                        }
                        gobble(inFASTA);
                }
+               
                inFASTA.close();
                outFASTA.close();
                scrapFASTA.close();
-               outGroups.close();
+               if (oligoFile != "") {   outGroups.close();   }
                if(qFileName != "")     {       qFile.close();  }
                
                for(int i=0;i<fastaFileNames.size();i++){
@@ -293,40 +441,107 @@ int TrimSeqsCommand::execute(){
                        delete fastaFileNames[i];
                }               
                
-               for(int i=0;i<fastaFileNames.size();i++){
-                       string seqName;
-                       openInputFile(getRootName(fastaFile) + groupVector[i] + ".fasta", inFASTA);
-                       ofstream outGroups;
-                       openOutputFile(outputDir + getRootName(getSimpleName(fastaFile)) + groupVector[i] + ".groups", outGroups);
-                       outputNames.push_back(outputDir + getRootName(getSimpleName(fastaFile)) + groupVector[i] + ".groups");
+               return 0;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimSeqsCommand", "driverCreateTrim");
+               exit(1);
+       }
+}
+/**************************************************************************************************/
+int TrimSeqsCommand::createProcessesCreateTrim(string filename, string qFileName, string trimFile, string scrapFile, string groupFile, vector<string> fastaNames) {
+       try {
+#if defined (__APPLE__) || (__MACH__) || (linux) || (__linux)
+               int process = 0;
+               int exitCommand = 1;
+               processIDS.clear();
+               
+               //loop through and create all the processes you want
+               while (process != processors) {
+                       int pid = fork();
                        
-                       while(!inFASTA.eof()){
-                               if(inFASTA.get() == '>'){
-                                       inFASTA >> seqName;
-                                       outGroups << seqName << '\t' << groupVector[i] << endl;
-                               }
-                               while (!inFASTA.eof())  {       char c = inFASTA.get(); if (c == 10 || c == 13){        break;  }       }
-                       }
-                       outGroups.close();
-                       inFASTA.close();
+                       if (pid > 0) {
+                               processIDS.push_back(pid);  //create map from line number to pid so you can append files in correct order later
+                               process++;
+                       }else if (pid == 0){
+                               driverCreateTrim(filename, qFileName, (trimFile + toString(getpid()) + ".temp"), (scrapFile + toString(getpid()) + ".temp"), (groupFile + toString(getpid()) + ".temp"), fastaNames, lines[process], qLines[process]);
+                               exit(0);
+                       }else { m->mothurOut("unable to spawn the necessary processes."); m->mothurOutEndLine(); exit(0); }
                }
                
-               m->mothurOutEndLine();
-               m->mothurOut("Output File Names: "); m->mothurOutEndLine();
-               for (int i = 0; i < outputNames.size(); i++) {  m->mothurOut(outputNames[i]); m->mothurOutEndLine();    }
-               m->mothurOutEndLine();
-                       
-               return 0;               
+               //force parent to wait until all the processes are done
+               for (int i=0;i<processors;i++) { 
+                       int temp = processIDS[i];
+                       wait(&temp);
+               }
+               
+               return exitCommand;
+#endif         
        }
        catch(exception& e) {
-               m->errorOut(e, "TrimSeqsCommand", "execute");
+               m->errorOut(e, "TrimSeqsCommand", "createProcessesCreateTrim");
                exit(1);
        }
 }
+/**************************************************************************************************/
+
+int TrimSeqsCommand::setLines(string filename, vector<linePair*>& lines) {
+       try {
+               
+               lines.clear();
+               
+               vector<long int> positions;
+               
+               ifstream inFASTA;
+               openInputFile(filename, inFASTA);
+                       
+               string input;
+               while(!inFASTA.eof()){  
+                       input = getline(inFASTA);
+
+                       if (input.length() != 0) {
+                               if(input[0] == '>'){ long int pos = inFASTA.tellg(); positions.push_back(pos - input.length() - 1);     }
+                       }
+               }
+               inFASTA.close();
+               
+               int numFastaSeqs = positions.size();
+       
+               FILE * pFile;
+               long size;
+               
+               //get num bytes in file
+               pFile = fopen (filename.c_str(),"rb");
+               if (pFile==NULL) perror ("Error opening file");
+               else{
+                       fseek (pFile, 0, SEEK_END);
+                       size=ftell (pFile);
+                       fclose (pFile);
+               }
+               
+               int numSeqsPerProcessor = numFastaSeqs / processors;
+               
+               for (int i = 0; i < processors; i++) {
 
+                       long int startPos = positions[ i * numSeqsPerProcessor ];
+                       if(i == processors - 1){
+                               numSeqsPerProcessor = numFastaSeqs - i * numSeqsPerProcessor;
+                       }else{  
+                               long int myEnd = positions[ (i+1) * numSeqsPerProcessor ];
+                       }
+                       lines.push_back(new linePair(startPos, numSeqsPerProcessor));
+               }
+               
+               return numFastaSeqs;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimSeqsCommand", "setLines");
+               exit(1);
+       }
+}
 //***************************************************************************************************************
 
-void TrimSeqsCommand::getOligos(vector<ofstream*>& outFASTAVec){
+void TrimSeqsCommand::getOligos(vector<string>& outFASTAVec){ //vector<ofstream*>& outFASTAVec
        try {
                ifstream inOligos;
                openInputFile(oligoFile, inOligos);
@@ -364,7 +579,9 @@ void TrimSeqsCommand::getOligos(vector<ofstream*>& outFASTAVec){
                                        groupVector.push_back(group);
                                        
                                        if(allFiles){
-                                               outFASTAVec.push_back(new ofstream((outputDir + getRootName(getSimpleName(fastaFile)) + group + ".fasta").c_str(), ios::ate));
+                                               //outFASTAVec.push_back(new ofstream((outputDir + getRootName(getSimpleName(fastaFile)) + group + ".fasta").c_str(), ios::ate));
+                                               outputNames.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + group + ".fasta"));
+                                               outFASTAVec.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + group + ".fasta"));
                                        }
                                }
                        }
@@ -380,9 +597,7 @@ void TrimSeqsCommand::getOligos(vector<ofstream*>& outFASTAVec){
                m->errorOut(e, "TrimSeqsCommand", "getOligos");
                exit(1);
        }
-
 }
-
 //***************************************************************************************************************
 
 bool TrimSeqsCommand::stripBarcode(Sequence& seq, int& group){
@@ -390,6 +605,7 @@ bool TrimSeqsCommand::stripBarcode(Sequence& seq, int& group){
                string rawSequence = seq.getUnaligned();
                bool success = 0;       //guilty until proven innocent
                
+               //can you find the barcode
                for(map<string,int>::iterator it=barcodes.begin();it!=barcodes.end();it++){
                        string oligo = it->first;
                        if(rawSequence.length() < oligo.length()){      //let's just assume that the barcodes are the same length
@@ -404,6 +620,48 @@ bool TrimSeqsCommand::stripBarcode(Sequence& seq, int& group){
                                break;
                        }
                }
+               
+               //if you found the barcode or if you don't want to allow for diffs
+               if ((bdiffs == 0) || (success == 1)) { return success;  }
+               
+               else { //try aligning and see if you can find it
+                       
+                       Alignment* alignment;
+                       if (barcodes.size() > 0) { //assumes barcodes are all the same length
+                               map<string,int>::iterator it=barcodes.begin();
+                               string temp = it->first;
+                               
+                               alignment = new NeedlemanOverlap(-2.0, 1.0, -1.0, (temp.length()+bdiffs+1));  
+                       }else{ alignment = NULL; } 
+                       
+
+                       //can you find the barcode
+                       for(map<string,int>::iterator it=barcodes.begin();it!=barcodes.end();it++){
+                               string oligo = it->first;
+                               int length = oligo.length();
+                               
+                               if(rawSequence.length() < oligo.length()){      //let's just assume that the barcodes are the same length
+                                       success = 0;
+                                       break;
+                               }
+                               
+                               //use needleman to align first barcode.length()+numdiffs of sequence to each barcode
+                               alignment->align(oligo, rawSequence.substr(0,length+bdiffs));
+                               oligo = alignment->getSeqAAln();
+                               string temp = alignment->getSeqBAln();
+               //cout << "barcode = " << oligo << " raw = " << rawSequence.substr(0,oligo.length()) << " raw aligned = " << temp << endl;                      
+                               
+                               int newStart=0;
+                               if(compareDNASeq(oligo, temp, length, newStart, bdiffs)){
+                                       group = it->second;
+                                       seq.setUnaligned(rawSequence.substr(newStart));
+                                       success = 1;
+                                       break;
+                               }
+                       }
+                       
+                       if (alignment != NULL) {  delete alignment;  }
+               }
                return success;
                
        }
@@ -436,6 +694,43 @@ bool TrimSeqsCommand::stripForward(Sequence& seq){
                        }
                }
                
+               //if you found the primer or if you don't want to allow for diffs
+               if ((pdiffs == 0) || (success == 1)) { return success;  }
+               
+               else { //try aligning and see if you can find it
+                       
+                       Alignment* alignment;
+                       if (numFPrimers > 0) {  alignment = new NeedlemanOverlap(-2.0, 1.0, -1.0, (forPrimer[0].length()+pdiffs+1));  } 
+                       else{ alignment = NULL; } 
+                       //can you find the primer
+                       for(int i=0;i<numFPrimers;i++){
+                               string oligo = forPrimer[i];
+                               int length = oligo.length();
+                       
+                               if(rawSequence.length() < oligo.length()){      
+                                       success = 0;
+                                       break;
+                               }
+                       
+                               //resize if neccessary
+                               if ((length+pdiffs+1) > alignment->getnRows()) { alignment->resize(length+pdiffs+1);    }
+                               
+                               //use needleman to align first primer.length()+numdiffs of sequence to each primer
+                               alignment->align(oligo, rawSequence.substr(0,length+pdiffs));
+                               oligo = alignment->getSeqAAln();
+                               string temp = alignment->getSeqBAln();
+                       
+                               int newStart = 0;
+                               if(compareDNASeq(oligo, temp, length, newStart, pdiffs)){
+                                       seq.setUnaligned(rawSequence.substr(newStart));
+                                       success = 1;
+                                       break;
+                               }
+                       }
+                       
+                       if (alignment != NULL) {  delete alignment;  }
+               }
+               
                return success;
                
        }
@@ -545,7 +840,7 @@ bool TrimSeqsCommand::compareDNASeq(string oligo, string seq){
                for(int i=0;i<length;i++){
                        
                        if(oligo[i] != seq[i]){
-                               if(oligo[i] == 'A' || oligo[i] == 'T' || oligo[i] == 'G' || oligo[i] == 'C')    {       success = 0;    }
+                               if(oligo[i] == 'A' || oligo[i] == 'T' || oligo[i] == 'G' || oligo[i] == 'C')    {       success = 0;    }
                                else if((oligo[i] == 'N' || oligo[i] == 'I') && (seq[i] == 'N'))                                {       success = 0;    }
                                else if(oligo[i] == 'R' && (seq[i] != 'A' && seq[i] != 'G'))                                    {       success = 0;    }
                                else if(oligo[i] == 'Y' && (seq[i] != 'C' && seq[i] != 'T'))                                    {       success = 0;    }
@@ -558,7 +853,7 @@ bool TrimSeqsCommand::compareDNASeq(string oligo, string seq){
                                else if(oligo[i] == 'H' && (seq[i] != 'A' && seq[i] != 'T' && seq[i] != 'C'))   {       success = 0;    }
                                else if(oligo[i] == 'V' && (seq[i] != 'A' && seq[i] != 'C' && seq[i] != 'G'))   {       success = 0;    }                       
                                
-                               if(success == 0)        {       break;  }
+                               if(success == 0)        {       break;   }
                        }
                        else{
                                success = 1;
@@ -573,16 +868,71 @@ bool TrimSeqsCommand::compareDNASeq(string oligo, string seq){
        }
 
 }
+//***************************************************************************************************************
 
+bool TrimSeqsCommand::compareDNASeq(string oligo, string seq, int numBases, int& end, int diffs){
+       try {
+               bool success = 1;
+               int length = oligo.length();
+               end = numBases;
+               int countBases = 0;
+               int countDiffs = 0;
+               
+               if (length != 0) {
+                       if ((oligo[0] == '-') || (oligo[0] == '.')) {  success = 0;  return success;  } //no gaps allowed at beginning
+               }
+               
+               for(int i=0;i<length;i++){
+                       
+                       if ((oligo[i] != '-') && (oligo[i] != '.'))  { countBases++; } 
+                                       
+                       if(oligo[i] != seq[i]){
+                               if(oligo[i] == 'A' || oligo[i] == 'T' || oligo[i] == 'G' || oligo[i] == 'C' || oligo[i] == '-' || oligo[i] == '.')      {       countDiffs++;   }
+                               else if((oligo[i] == 'N' || oligo[i] == 'I') && (seq[i] == 'N'))                                {       countDiffs++;   }
+                               else if(oligo[i] == 'R' && (seq[i] != 'A' && seq[i] != 'G'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'Y' && (seq[i] != 'C' && seq[i] != 'T'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'M' && (seq[i] != 'C' && seq[i] != 'A'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'K' && (seq[i] != 'T' && seq[i] != 'G'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'W' && (seq[i] != 'T' && seq[i] != 'A'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'S' && (seq[i] != 'C' && seq[i] != 'G'))                                    {       countDiffs++;   }
+                               else if(oligo[i] == 'B' && (seq[i] != 'C' && seq[i] != 'T' && seq[i] != 'G'))   {       countDiffs++;   }
+                               else if(oligo[i] == 'D' && (seq[i] != 'A' && seq[i] != 'T' && seq[i] != 'G'))   {       countDiffs++;   }
+                               else if(oligo[i] == 'H' && (seq[i] != 'A' && seq[i] != 'T' && seq[i] != 'C'))   {       countDiffs++;   }
+                               else if(oligo[i] == 'V' && (seq[i] != 'A' && seq[i] != 'C' && seq[i] != 'G'))   {       countDiffs++;   }                       
+                               
+                               if(countDiffs > diffs)  {       success = 0; break;      }
+                       }
+                       else{
+                               success = 1;
+                       }
+                       
+                       if (countBases >= numBases) { end = countBases; break; } //stop checking after end of barcode or primer
+               }
+               
+               //if it's a success we want to check for total diffs in driver, so save it.
+               if (success == 1) {  currentSeqsTdiffs = countDiffs; }
+               
+               return success;
+       }
+       catch(exception& e) {
+               m->errorOut(e, "TrimSeqsCommand", "compareDNASeq");
+               exit(1);
+       }
+
+}
 //***************************************************************************************************************
 
 bool TrimSeqsCommand::stripQualThreshold(Sequence& seq, ifstream& qFile){
        try {
                string rawSequence = seq.getUnaligned();
-               int seqLength = rawSequence.length();
-               string name;
+               int seqLength;  // = rawSequence.length();
+               string name, temp, temp2;
                
-               qFile >> name;
+               qFile >> name >> temp;
+       
+               splitAtEquals(temp2, temp); //separates length=242, temp=length, temp2=242
+               convert(temp, seqLength); //converts string to int
+       
                if (name.length() != 0) {  if(name.substr(1) != seq.getName())  {       m->mothurOut("sequence name mismatch btwn fasta and qual file"); m->mothurOutEndLine(); }  } 
                while (!qFile.eof())    {       char c = qFile.get(); if (c == 10 || c == 13){  break;  }       }
                
@@ -621,7 +971,7 @@ bool TrimSeqsCommand::cullQualAverage(Sequence& seq, ifstream& qFile){
                string name;
                
                qFile >> name;
-               if (name[0] == '>') {  if(name.substr(1) != seq.getName())      {       m->mothurOut("sequence name mismatch btwn fasta: " + seq.getName() + " and qual file: " + name); m->mothurOutEndLine(); } }
+               if (name[0] == '>') {  if(name.substr(1) != seq.getName())      {       m->mothurOut("sequence name mismatch btwn fasta: " + seq.getName() + " and qual file: " + name); m->mothurOutEndLine(); }  }
                
                while (!qFile.eof())    {       char c = qFile.get(); if (c == 10 || c == 13){  break;  }       }