try {
abort = false;
+ comboStarts = 0;
//allow user to run help
if(option == "help") { help(); abort = true; }
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, 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 flip parameter will output the reverse compliment of your trimmed sequence. The default is false.\n");
m->mothurOut("The oligos parameter .... The default is "".\n");
m->mothurOut("The maxambig parameter .... The default is -1.\n");
m->mothurOut("The maxhomop parameter .... The default is 0.\n");
#if defined (__APPLE__) || (__MACH__) || (linux) || (__linux)
if(processors == 1){
ifstream inFASTA;
+ int numSeqs;
openInputFile(fastaFile, inFASTA);
- int numSeqs=count(istreambuf_iterator<char>(inFASTA),istreambuf_iterator<char>(), '>');
+ getNumSeqs(inFASTA, numSeqs);
inFASTA.close();
lines.push_back(new linePair(0, numSeqs));
if (m->control_pressed) { return 0; }
#else
ifstream inFASTA;
- openInputFile(fastafileNames[s], inFASTA);
- numSeqs=count(istreambuf_iterator<char>(inFASTA),istreambuf_iterator<char>(), '>');
+ int numSeqs;
+ openInputFile(fastaFile, inFASTA);
+ getNumSeqs(inFASTA, numSeqs);
inFASTA.close();
lines.push_back(new linePair(0, numSeqs));
- driverCreateSummary(fastafile, qFileName, trimSeqFile, scrapSeqFile, groupFile, fastaFileNames, lines[0], lines[0]);
+ driverCreateTrim(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;
+ if (isBlank(fastaFileNames[i])) { remove(fastaFileNames[i].c_str()); }
+ else if (filesToRemove.count(fastaFileNames[i]) > 0) { remove(fastaFileNames[i].c_str()); }
+ else {
+ ifstream inFASTA;
+ string seqName;
+ //openInputFile(getRootName(fastaFile) + groupVector[i] + ".fasta", inFASTA);
+ openInputFile(fastaFileNames[i], inFASTA);
+ ofstream outGroups;
+ string outGroupFilename = outputDir + getRootName(getSimpleName(fastaFileNames[i])) + "groups";
+ openOutputFile(outGroupFilename, outGroups);
+ //openOutputFile(outputDir + getRootName(getSimpleName(fastaFile)) + groupVector[i] + ".groups", outGroups);
+ outputNames.push_back(outGroupFilename);
+
+ string thisGroup = "";
+ if (i > comboStarts) {
+ map<string, int>::iterator itCombo;
+ for(itCombo=combos.begin();itCombo!=combos.end(); itCombo++){
+ if(itCombo->second == i){ thisGroup = itCombo->first; combos.erase(itCombo); break; }
+ }
+ }else{ thisGroup = groupVector[i]; }
+
+ while(!inFASTA.eof()){
+ if(inFASTA.get() == '>'){
+ inFASTA >> seqName;
+ outGroups << seqName << '\t' << thisGroup << endl;
+ }
+ while (!inFASTA.eof()) { char c = inFASTA.get(); if (c == 10 || c == 13){ break; } }
}
- while (!inFASTA.eof()) { char c = inFASTA.get(); if (c == 10 || c == 13){ break; } }
+ outGroups.close();
+ inFASTA.close();
}
- outGroups.close();
- inFASTA.close();
}
if (m->control_pressed) {
if (oligoFile != "") {
openOutputFile(groupFile, outGroups);
for (int i = 0; i < fastaNames.size(); i++) {
+ #if defined (__APPLE__) || (__MACH__) || (linux) || (__linux)
fastaFileNames.push_back(new ofstream((fastaNames[i] + toString(getpid()) + ".temp").c_str(), ios::ate));
+ #else
+ fastaFileNames.push_back(new ofstream((fastaNames[i] + toString(i) + ".temp").c_str(), ios::ate));
+ #endif
}
}
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];
- }
+ 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;
+ int success = 1;
Sequence currSeq(inFASTA);
+
string origSeq = currSeq.getUnaligned();
if (origSeq != "") {
- int group;
+ int groupBar, groupPrime;
string trashCode = "";
int currentSeqsDiffs = 0;
if(qFileName != ""){
if(qThreshold != 0) { success = stripQualThreshold(currSeq, qFile); }
else if(qAverage != 0) { success = cullQualAverage(currSeq, qFile); }
- if ((!qtrim) && (origSeq.length() != currSeq.getUnaligned().length())) {
+
+ if (qtrim == 1 && (origSeq.length() != currSeq.getUnaligned().length())) {
success = 0; //if you don't want to trim and the sequence does not meet quality requirements, move to scrap
}
+
if(!success) { trashCode += 'q'; }
}
if(barcodes.size() != 0){
- success = stripBarcode(currSeq, group);
-// cout << "here: " << success << endl;
+ success = stripBarcode(currSeq, groupBar);
if(success > bdiffs){ trashCode += 'b'; }
else{ currentSeqsDiffs += success; }
}
if(numFPrimers != 0){
- success = stripForward(currSeq);
+ success = stripForward(currSeq, groupPrime);
if(success > pdiffs){ trashCode += 'f'; }
else{ currentSeqsDiffs += success; }
}
if(flip){ currSeq.reverseComplement(); } // should go last
if(trashCode.length() == 0){
- currSeq.setAligned(currSeq.getUnaligned()); //this is because of a modification we made to the sequence class to fix a bug. all seqs have an aligned version, which is the version that gets printed.
+ currSeq.setAligned(currSeq.getUnaligned());
currSeq.printSequence(outFASTA);
if(barcodes.size() != 0){
- outGroups << currSeq.getName() << '\t' << groupVector[group] << endl;
-
+ string thisGroup = groupVector[groupBar];
+ int indexToFastaFile = groupBar;
+ if (primers.size() != 0){
+ //does this primer have a group
+ if (groupVector[groupPrime] != "") {
+ thisGroup += "." + groupVector[groupPrime];
+ indexToFastaFile = combos[thisGroup];
+ }
+ }
+ outGroups << currSeq.getName() << '\t' << thisGroup << endl;
if(allFiles){
- currSeq.printSequence(*fastaFileNames[group]);
+ currSeq.printSequence(*fastaFileNames[indexToFastaFile]);
}
}
}
else{
currSeq.setName(currSeq.getName() + '|' + trashCode);
currSeq.setUnaligned(origSeq);
+ currSeq.setAligned(origSeq);
currSeq.printSequence(scrapFASTA);
}
}
string type, oligo, group;
int index=0;
+ //int indexPrimer = 0;
while(!inOligos.eof()){
inOligos >> type;
-
+
if(type[0] == '#'){
while (!inOligos.eof()) { char c = inOligos.get(); if (c == 10 || c == 13){ break; } } // get rest of line if there's any crap there
}
else{
+ //make type case insensitive
+ for(int i=0;i<type.length();i++){ type[i] = toupper(type[i]); }
+
inOligos >> oligo;
for(int i=0;i<oligo.length();i++){
if(oligo[i] == 'U') { oligo[i] = 'T'; }
}
- if(type == "forward"){
- forPrimer.push_back(oligo);
+ if(type == "FORWARD"){
+ group = "";
+
+ // get rest of line in case there is a primer name
+ while (!inOligos.eof()) {
+ char c = inOligos.get();
+ if (c == 10 || c == 13){ break; }
+ else if (c == 32 || c == 9){;} //space or tab
+ else { group += c; }
+ }
+
+ //check for repeat barcodes
+ map<string, int>::iterator itPrime = primers.find(oligo);
+ if (itPrime != primers.end()) { m->mothurOut("primer " + oligo + " is in your oligos file already."); m->mothurOutEndLine(); }
+
+ primers[oligo]=index; index++;
+ groupVector.push_back(group);
+
+ if(allFiles){
+ outFASTAVec.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(index) + "." + group + ".fasta"));
+ if (group == "") { //if there is not a group for this primer, then this file will not get written to, but we add it to keep the indexes correct
+ filesToRemove.insert((outputDir + getRootName(getSimpleName(fastaFile)) + toString(index) + "." + group + ".fasta"));
+ }else {
+ outputNames.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(index) + "." + group + ".fasta"));
+ }
+ }
+
}
- else if(type == "reverse"){
+ else if(type == "REVERSE"){
Sequence oligoRC("reverse", oligo);
oligoRC.reverseComplement();
revPrimer.push_back(oligoRC.getUnaligned());
}
- else if(type == "barcode"){
+ else if(type == "BARCODE"){
inOligos >> group;
- barcodes[oligo]=index++;
+
+ //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]=index; index++;
groupVector.push_back(group);
if(allFiles){
- //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"));
+ outputNames.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(index) + "." + group + ".fasta"));
+ outFASTAVec.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(index) + "." + group + ".fasta"));
}
- }
+ }else{ m->mothurOut(type + " is not recognized as a valid type. Choices are forward, reverse, and barcode. Ignoring " + oligo + "."); m->mothurOutEndLine(); }
}
+ gobble(inOligos);
}
inOligos.close();
- numFPrimers = forPrimer.size();
+ //add in potential combos
+ if(allFiles){
+ comboStarts = outFASTAVec.size()-1;
+ for (map<string, int>::iterator itBar = barcodes.begin(); itBar != barcodes.end(); itBar++) {
+ for (map<string, int>::iterator itPrime = primers.begin(); itPrime != primers.end(); itPrime++) {
+ if (groupVector[itPrime->second] != "") { //there is a group for this primer
+ outputNames.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(itBar->second) + "." + groupVector[itBar->second] + "." + toString(itPrime->second) + "." + groupVector[itPrime->second] + ".fasta"));
+ outFASTAVec.push_back((outputDir + getRootName(getSimpleName(fastaFile)) + toString(itBar->second) + "." + groupVector[itBar->second] + "." + toString(itPrime->second) + "." + groupVector[itPrime->second] + ".fasta"));
+ combos[(groupVector[itBar->second] + "." + groupVector[itPrime->second])] = outFASTAVec.size()-1;
+ }
+ }
+ }
+ }
+
+ numFPrimers = primers.size();
numRPrimers = revPrimer.size();
}
int TrimSeqsCommand::stripBarcode(Sequence& seq, int& group){
try {
+
string rawSequence = seq.getUnaligned();
- bool success = bdiffs + 1; //guilty until proven innocent
+ int success = bdiffs + 1; //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
- success = bdiffs + 1;
- break;
+ success = bdiffs + 10; //if the sequence is shorter than the barcode then bail out
+ break;
}
if(compareDNASeq(oligo, rawSequence.substr(0,oligo.length()))){
maxLength = it->first.length();
}
}
- alignment = new NeedlemanOverlap(-2.0, 1.0, -1.0, (maxLength+bdiffs+1));
+ alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxLength+bdiffs+1));
}else{ alignment = NULL; }
// int length = oligo.length();
if(rawSequence.length() < maxLength){ //let's just assume that the barcodes are the same length
- success = bdiffs + 1;
+ success = bdiffs + 10;
break;
}
}
oligo = oligo.substr(0,alnLength);
temp = temp.substr(0,alnLength);
-// cout << "barcode = " << oligo << " raw = " << rawSequence.substr(0,alnLength) << " raw aligned = " << temp << endl;
- cout << seq.getName() << endl;
- cout << temp << endl;
- cout << oligo << endl;
- cout << alnLength << endl;
- cout << endl;
int newStart=0;
int numDiff = countDiffs(oligo, temp);
+
+// cout << oligo << '\t' << temp << '\t' << numDiff << endl;
+
if(numDiff < minDiff){
minDiff = numDiff;
minCount = 1;
}
}
- if(minDiff > bdiffs){ success = bdiffs + 1; }
- else if(minCount > 1) { success = bdiffs + 1; }
- else{
+
+ if(minDiff > bdiffs) { success = minDiff; } //no good matches
+ else if(minCount > 1) { success = bdiffs + 100; } //can't tell the difference between multiple barcodes
+ else{ //use the best match
group = minGroup;
- seq.setUnaligned("*" + rawSequence.substr(minPos));
+ seq.setUnaligned(rawSequence.substr(minPos));
success = minDiff;
}
if (alignment != NULL) { delete alignment; }
}
+// cout << success << endl;
+
return success;
}
//***************************************************************************************************************
-int TrimSeqsCommand::stripForward(Sequence& seq){
+int TrimSeqsCommand::stripForward(Sequence& seq, int& group){
try {
string rawSequence = seq.getUnaligned();
- bool success = pdiffs + 1; //guilty until proven innocent
+ int success = pdiffs + 1; //guilty until proven innocent
//can you find the primer
- for(int i=0;i<numFPrimers;i++){
- string oligo = forPrimer[i];
-
- if(rawSequence.length() < oligo.length()){
- success = pdiffs + 1;
- break;
+ for(map<string,int>::iterator it=primers.begin();it!=primers.end();it++){
+ string oligo = it->first;
+ if(rawSequence.length() < oligo.length()){ //let's just assume that the primers are the same length
+ success = pdiffs + 10; //if the sequence is shorter than the barcode then bail out
+ break;
}
if(compareDNASeq(oligo, rawSequence.substr(0,oligo.length()))){
+ group = it->second;
seq.setUnaligned(rawSequence.substr(oligo.length()));
success = 0;
break;
}
}
-
+
//if you found the barcode or if you don't want to allow for diffs
// cout << success;
if ((pdiffs == 0) || (success == 0)) { return success; }
int maxLength = 0;
Alignment* alignment;
- if (numFPrimers > 0) {
+ if (primers.size() > 0) {
+ map<string,int>::iterator it=primers.begin();
- for(int i=0;i<numFPrimers;i++){
- if(forPrimer[i].length() > maxLength){
- maxLength = forPrimer[i].length();
+ for(it;it!=primers.end();it++){
+ if(it->first.length() > maxLength){
+ maxLength = it->first.length();
}
}
- alignment = new NeedlemanOverlap(-2.0, 1.0, -1.0, (maxLength+pdiffs+1));
+ alignment = new NeedlemanOverlap(-1.0, 1.0, -1.0, (maxLength+pdiffs+1));
}else{ alignment = NULL; }
//can you find the barcode
int minDiff = 1e6;
int minCount = 1;
+ int minGroup = -1;
int minPos = 0;
- for(int i=0;i<numFPrimers;i++){
- string oligo = forPrimer[i];
+ for(map<string,int>::iterator it=primers.begin();it!=primers.end();it++){
+ string oligo = it->first;
+// int length = oligo.length();
if(rawSequence.length() < maxLength){
- success = pdiffs + 1;
+ success = pdiffs + 100;
break;
}
}
oligo = oligo.substr(0,alnLength);
temp = temp.substr(0,alnLength);
-// cout << "barcode = " << oligo << " raw = " << rawSequence.substr(0,alnLength) << " raw aligned = " << temp << endl;
- cout << seq.getName() << endl;
- cout << temp << endl;
- cout << oligo << endl;
- cout << alnLength << endl;
- cout << endl;
int newStart=0;
int numDiff = countDiffs(oligo, temp);
+
+// cout << oligo << '\t' << temp << '\t' << numDiff << endl;
+
if(numDiff < minDiff){
minDiff = numDiff;
minCount = 1;
+ minGroup = it->second;
minPos = 0;
for(int i=0;i<alnLength;i++){
if(temp[i] != '-'){
}
}
- if(minDiff > pdiffs){ success = pdiffs + 1; }
- else if(minCount > 1) { success = pdiffs + 1; }
- else{
- seq.setUnaligned("*" + rawSequence.substr(minPos));
+
+ if(minDiff > pdiffs) { success = minDiff; } //no good matches
+ else if(minCount > 1) { success = pdiffs + 10; } //can't tell the difference between multiple primers
+ else{ //use the best match
+ group = minGroup;
+ seq.setUnaligned(rawSequence.substr(minPos));
success = minDiff;
}
if (alignment != NULL) { delete alignment; }
}
+
return success;
}
bool TrimSeqsCommand::stripQualThreshold(Sequence& seq, ifstream& qFile){
try {
+// string rawSequence = seq.getUnaligned();
+// int seqLength; // = rawSequence.length();
+// string name, temp, temp2;
+//
+// qFile >> name;
+//
+// //get rest of line
+// temp = "";
+// while (!qFile.eof()) {
+// char c = qFile.get();
+// if (c == 10 || c == 13){ break; }
+// else { temp += c; }
+// }
+//
+// int pos = temp.find("length");
+// if (pos == temp.npos) { m->mothurOut("Cannot find length in qfile for " + seq.getName()); m->mothurOutEndLine(); seqLength = 0; }
+// else {
+// string tempLength = temp.substr(pos);
+// istringstream iss (tempLength,istringstream::in);
+// iss >> 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(); } }
+
string rawSequence = seq.getUnaligned();
- int seqLength; // = rawSequence.length();
- string name, temp, temp2;
+ int seqLength = seq.getNumBases();
+ bool success = 0; //guilty until proven innocent
+ 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(); } }
- 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; } }
int score;
for(int i=0;i<seqLength;i++){
qFile >> score;
- if(score <= qThreshold){
+ if(score < qThreshold){
end = i;
break;
}