CommandParameter pstart("start", "Number", "", "-1", "", "", "","",false,false); parameters.push_back(pstart);
CommandParameter pend("end", "Number", "", "-1", "", "", "","",false,false); parameters.push_back(pend);
CommandParameter pnomatch("nomatch", "Multiple", "reject-keep", "reject", "", "", "","",false,false); parameters.push_back(pnomatch);
+ CommandParameter ppdiffs("pdiffs", "Number", "", "0", "", "", "","",false,false,true); parameters.push_back(ppdiffs);
+
CommandParameter pprocessors("processors", "Number", "", "1", "", "", "","",false,false,true); parameters.push_back(pprocessors);
CommandParameter pkeepprimer("keepprimer", "Boolean", "", "F", "", "", "","",false,false); parameters.push_back(pkeepprimer);
CommandParameter pkeepdots("keepdots", "Boolean", "", "T", "", "", "","",false,false); parameters.push_back(pkeepdots);
try {
string helpString = "";
helpString += "The pcr.seqs command reads a fasta file.\n";
- helpString += "The pcr.seqs command parameters are fasta, oligos, name, group, count, taxonomy, ecoli, start, end, nomatch, processors, keepprimer and keepdots.\n";
+ helpString += "The pcr.seqs command parameters are fasta, oligos, name, group, count, taxonomy, ecoli, start, end, nomatch, pdiffs, processors, keepprimer and keepdots.\n";
helpString += "The ecoli parameter is used to provide a fasta file containing a single reference sequence (e.g. for e. coli) this must be aligned. Mothur will trim to the start and end positions of the reference sequence.\n";
helpString += "The start parameter allows you to provide a starting position to trim to.\n";
helpString += "The end parameter allows you to provide a ending position to trim from.\n";
helpString += "The processors parameter allows you to use multiple processors.\n";
helpString += "The keepprimer parameter allows you to keep the primer, default=false.\n";
helpString += "The keepdots parameter allows you to keep the leading and trailing .'s, default=true.\n";
+ helpString += "The pdiffs parameter is used to specify the number of differences allowed in the primer. The default is 0.\n";
helpString += "Note: No spaces between parameter labels (i.e. fasta), '=' and parameters (i.e.yourFasta).\n";
helpString += "For more details please check out the wiki http://www.mothur.org/wiki/Pcr.seqs .\n";
return helpString;
temp = validParameter.validFile(parameters, "processors", false); if (temp == "not found"){ temp = m->getProcessors(); }
m->setProcessors(temp);
m->mothurConvert(temp, processors);
+
+ temp = validParameter.validFile(parameters, "pdiffs", false); if (temp == "not found") { temp = "0"; }
+ m->mothurConvert(temp, pdiffs);
nomatch = validParameter.validFile(parameters, "nomatch", false); if (nomatch == "not found") { nomatch = "reject"; }
length = 0;
- if(oligosfile != ""){ readOligos(); } if (m->control_pressed) { return 0; }
+ if(oligosfile != ""){ readOligos(); if (m->debug) { m->mothurOut("[DEBUG]: read oligos file. numprimers = " + toString(primers.size()) + ", revprimers = " + toString(revPrimer.size()) + ".\n"); } } if (m->control_pressed) { return 0; }
if(ecolifile != "") { readEcoli(); } if (m->control_pressed) { return 0; }
vector<unsigned long long> positions;
if (i!=0) {extension += toString(i) + ".temp"; processIDS.push_back(i); }
// Allocate memory for thread data.
- pcrData* tempPcr = new pcrData(filename, goodFileName+extension, badFileName+extension, m, oligosfile, ecolifile, primers, revPrimer, nomatch, keepprimer, keepdots, start, end, length, lines[i].start, lines[i].end);
+ pcrData* tempPcr = new pcrData(filename, goodFileName+extension, badFileName+extension, m, oligosfile, ecolifile, primers, revPrimer, nomatch, keepprimer, keepdots, start, end, length, pdiffs, lines[i].start, lines[i].end);
pDataArray.push_back(tempPcr);
//default security attributes, thread function name, argument to thread function, use default creation flags, returns the thread identifier
int count = 0;
set<int> lengths;
+ //pdiffs, bdiffs, primers, barcodes, revPrimers
+ map<string, int> faked;
+ TrimOligos trim(pdiffs, 0, primers, faked, revPrimer);
+
while (!done) {
if (m->control_pressed) { break; }
string trashCode = "";
if (currSeq.getName() != "") {
+ if (m->debug) { m->mothurOut("[DEBUG]: seq name = " + currSeq.getName() + ".\n"); }
+
bool goodSeq = true;
if (oligosfile != "") {
map<int, int> mapAligned;
//process primers
if (primers.size() != 0) {
int primerStart = 0; int primerEnd = 0;
- bool good = findForward(currSeq, primerStart, primerEnd);
+ bool good = trim.findForward(currSeq, primerStart, primerEnd);
if(!good){ if (nomatch == "reject") { goodSeq = false; } trashCode += "f"; }
else{
}
else {
if (keepdots) { currSeq.filterToPos(mapAligned[primerStart]); }
- else { currSeq.setAligned(currSeq.getAligned().substr(mapAligned[primerStart])); }
+ else { currSeq.setAligned(currSeq.getAligned().substr(mapAligned[primerStart])); }
}
+ isAligned(currSeq.getAligned(), mapAligned);
}else {
if (!keepprimer) { currSeq.setAligned(currSeq.getUnaligned().substr(primerEnd)); }
else { currSeq.setAligned(currSeq.getUnaligned().substr(primerStart)); }
//process reverse primers
if (revPrimer.size() != 0) {
int primerStart = 0; int primerEnd = 0;
- bool good = findReverse(currSeq, primerStart, primerEnd);
+ bool good = trim.findReverse(currSeq, primerStart, primerEnd);
if(!good){ if (nomatch == "reject") { goodSeq = false; } trashCode += "r"; }
else{
- //are you aligned
+ //are you aligned
if (aligned) {
if (!keepprimer) {
if (keepdots) { currSeq.filterFromPos(mapAligned[primerStart]); }
}
}
//********************************************************************/
-bool PcrSeqsCommand::findForward(Sequence& seq, int& primerStart, int& primerEnd){
- try {
-
- string rawSequence = seq.getUnaligned();
-
- for(int j=0;j<primers.size();j++){
- string oligo = primers[j];
-
- 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;
- return false;
-
- }
- catch(exception& e) {
- m->errorOut(e, "TrimOligos", "stripForward");
- exit(1);
- }
-}
-//******************************************************************/
-bool PcrSeqsCommand::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);
- }
-}
-//********************************************************************/
bool PcrSeqsCommand::isAligned(string seq, map<int, int>& aligned){
try {
+ aligned.clear();
bool isAligned = false;
int countBases = 0;
m->openInputFile(oligosfile, inOligos);
string type, oligo, group;
+ int primerCount = 0;
while(!inOligos.eof()){
if (c == 10 || c == 13){ break; }
else if (c == 32 || c == 9){;} //space or tab
}
- primers.push_back(oligo);
+ primers[oligo] = primerCount; primerCount++;
}else if(type == "REVERSE"){
string oligoRC = reverseOligo(oligo);
revPrimer.push_back(oligoRC);
exit(1);
}
-}
-//******************************************************************/
-bool PcrSeqsCommand::compareDNASeq(string oligo, string seq){
- try {
- bool success = 1;
- int length = oligo.length();
-
- 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; }
- 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; }
- else if(oligo[i] == 'M' && (seq[i] != 'C' && seq[i] != 'A')) { success = 0; }
- else if(oligo[i] == 'K' && (seq[i] != 'T' && seq[i] != 'G')) { success = 0; }
- else if(oligo[i] == 'W' && (seq[i] != 'T' && seq[i] != 'A')) { success = 0; }
- else if(oligo[i] == 'S' && (seq[i] != 'C' && seq[i] != 'G')) { success = 0; }
- else if(oligo[i] == 'B' && (seq[i] != 'C' && seq[i] != 'T' && seq[i] != 'G')) { success = 0; }
- else if(oligo[i] == 'D' && (seq[i] != 'A' && seq[i] != 'T' && seq[i] != 'G')) { success = 0; }
- 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; }
- }
- else{
- success = 1;
- }
- }
-
- return success;
- }
- catch(exception& e) {
- m->errorOut(e, "PcrSeqsCommand", "compareDNASeq");
- exit(1);
- }
-
}
//***************************************************************************************************************
int PcrSeqsCommand::readName(set<string>& names){