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/*
* decalc.cpp
* Mothur
*
* Created by Sarah Westcott on 7/22/09.
* Copyright 2009 Schloss Lab UMASS Amherst. All rights reserved.
*
*/
#include "decalc.h"
#include "mothurchimera.h"
#include "calculator.h"
#include "eachgapdist.h"
#include "ignoregaps.h"
#include "eachgapdist.h"
//***************************************************************************************************************
void DeCalculator::setMask(string ms) {
try {
seqMask = ms;
int count = 0;
maskMap.clear();
if (seqMask.length() != 0) {
//whereever there is a base in the mask, save that value is query and subject
for (int i = 0; i < seqMask.length(); i++) {
if (isalpha(seqMask[i])) {
h.insert(i);
maskMap[count] = i;
count++;
}
}
}else {
for (int i = 0; i < alignLength; i++) {
h.insert(i);
maskMap[count] = i;
count++;
}
}
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "setMask");
exit(1);
}
}
//***************************************************************************************************************
void DeCalculator::runMask(Sequence* seq) {
try{
string q = seq->getAligned();
string tempQuery = "";
//whereever there is a base in the mask, save that value is query and subject
set<int>::iterator setit;
for ( setit=h.begin() ; setit != h.end(); setit++ ) {
tempQuery += q[*setit];
}
//save masked values
seq->setAligned(tempQuery);
seq->setUnaligned(tempQuery);
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "runMask");
exit(1);
}
}
//***************************************************************************************************************
//num is query's spot in querySeqs
void DeCalculator::trimSeqs(Sequence* query, Sequence* subject, map<int, int>& trim) {
try {
string q = query->getAligned();
string s = subject->getAligned();
int front = 0;
for (int i = 0; i < q.length(); i++) { if (isalpha(q[i]) && isalpha(s[i])) { front = i; break; } }
int back = 0;
for (int i = q.length(); i >= 0; i--) { if (isalpha(q[i]) && isalpha(s[i])) { back = i; break; } }
trim[front] = back;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "trimSeqs");
exit(1);
}
}
//***************************************************************************************************************
vector<int> DeCalculator::findWindows(Sequence* query, int front, int back, int& size, int increment) {
try {
vector<int> win;
int cutoff = back - front; //back - front
//if window is set to default
if (size == 0) { if (cutoff > 1200) { size = 300; }
else{ size = (cutoff / 4); } }
else if (size > (cutoff / 4)) {
m->mothurOut("[WARNING]: You have selected too large a window size for sequence " + query->getName() + ". I will choose an appropriate window size.\n");
size = (cutoff / 4);
}
//this follows wigeon, but we may want to consider that it chops off the end values if the sequence cannot be evenly divided into steps
for (int i = front; i < (back - size) ; i+=increment) { win.push_back(i); }
return win;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "findWindows");
exit(1);
}
}
//***************************************************************************************************************
vector<float> DeCalculator::calcObserved(Sequence* query, Sequence* subject, vector<int> window, int size) {
try {
if (query->getAligned().length() != subject->getAligned().length()) {
m->mothurOut("[ERROR]: seqLengths must match. queryLength = " + toString(query->getAligned().length()) + ", subjectLength = " + toString(subject->getAligned().length()) +"\n");
m->setControl_pressed(true);
}
vector<float> temp;
//int gaps = 0;
for (int i = 0; i < window.size(); i++) {
if (m->getControl_pressed()) { break; }
string seqFrag = query->getAligned().substr(window[i], size);
string seqFragsub = subject->getAligned().substr(window[i], size);
if (m->getDebug()) { m->mothurOut("[DEBUG]: seqLengths = " + toString(seqFrag.length()) + "," + toString(seqFragsub.length()) + "\t" + toString(window[i]) + "\t" + toString(size) +"\n"); }
int diff = 0;
for (int b = 0; b < seqFrag.length(); b++) {
//if at least one is a base and they are not equal
if( (isalpha(seqFrag[b]) || isalpha(seqFragsub[b])) && (seqFrag[b] != seqFragsub[b]) ) { diff++; }
}
//percentage of mismatched bases
float dist = diff / (float) (seqFrag.length()) * 100;
if (m->getDebug()) { m->mothurOut("[DEBUG]: diffs = " + toString(diff) + ", dist = " + toString(dist) + "\n"); }
temp.push_back(dist);
}
return temp;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "calcObserved");
exit(1);
}
}
//***************************************************************************************************************
float DeCalculator::calcDist(Sequence* query, Sequence* subject, int front, int back) {
try {
//so you only look at the trimmed part of the sequence
int cutoff = back - front;
int gaps = 0;
//from first startpoint with length back-front
string seqFrag = query->getAligned().substr(front, cutoff);
string seqFragsub = subject->getAligned().substr(front, cutoff);
int diff = 0;
for (int b = 0; b < seqFrag.length(); b++) {
//ignore gaps
if((!isalpha(seqFrag[b])) && (!isalpha(seqFragsub[b]))) { gaps++; }
if (seqFrag[b] != seqFragsub[b]) { diff++; }
}
//if the whole fragment is 0 distance = 0
if ((seqFrag.length()-gaps) == 0) { return 0.0; }
//percentage of mismatched bases
float dist = diff / (float) (seqFrag.length()-gaps) * 100;
return dist;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "calcDist");
exit(1);
}
}
//***************************************************************************************************************
vector<float> DeCalculator::calcExpected(vector<float> qav, float coef) {
try {
//for each window
vector<float> queryExpected;
for (int j = 0; j < qav.size(); j++) {
float expected = qav[j] * coef;
queryExpected.push_back(expected);
}
return queryExpected;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "calcExpected");
exit(1);
}
}
//***************************************************************************************************************
float DeCalculator::calcDE(vector<float> obs, vector<float> exp) {
try {
//for each window
float sum = 0.0; //sum = sum from 1 to i of (oi-ei)^2
int numZeros = 0;
for (int j = 0; j < obs.size(); j++) { sum += ((obs[j] - exp[j]) * (obs[j] - exp[j])); }
float de = sqrt((sum / (obs.size() - 1 - numZeros)));
return de;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "calcDE");
exit(1);
}
}
//***************************************************************************************************************
vector<float> DeCalculator::calcFreq(vector<Sequence*> seqs, string filename, string version) {
try {
vector<float> prob;
Utils util;
string freqfile = util.getRootName(filename) + "freq";
ofstream outFreq;
util.openOutputFile(freqfile, outFreq);
outFreq << "#" << version << endl;
string length = toString(seqs.size()); //if there are 5000 seqs in the template then set precision to 3
int precision = length.length() - 1;
//format output
outFreq.setf(ios::fixed, ios::floatfield); outFreq.setf(ios::showpoint);
//at each position in the sequence
for (int i = 0; i < seqs[0]->getAligned().length(); i++) {
vector<int> freq; freq.resize(4,0);
int gaps = 0;
//find the frequency of each nucleotide
for (int j = 0; j < seqs.size(); j++) {
char value = seqs[j]->getAligned()[i];
if(toupper(value) == 'A') { freq[0]++; }
else if(toupper(value) == 'T' || toupper(value) == 'U') { freq[1]++; }
else if(toupper(value) == 'G') { freq[2]++; }
else if(toupper(value) == 'C') { freq[3]++; }
else { gaps++; }
}
//find base with highest frequency
int highest = 0;
for (int j = 0; j < freq.size(); j++) { if (freq[j] > highest) { highest = freq[j]; } }
float highFreq = highest / (float) (seqs.size());
float Pi;
Pi = (highFreq - 0.25) / 0.75;
//cannot have probability less than 0.
if (Pi < 0) { Pi = 0.0; }
//saves this for later
outFreq << setprecision(precision) << i << '\t' << highFreq << endl;
if (h.count(i) > 0) {
prob.push_back(Pi);
}
}
outFreq.close();
return prob;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "calcFreq");
exit(1);
}
}
//***************************************************************************************************************
vector<float> DeCalculator::findQav(vector<int> window, int size, vector<float> probabilityProfile) {
try {
vector<float> averages;
//for each window find average
for (int i = 0; i < window.size(); i++) {
float average = 0.0;
//while you are in the window for this sequence
int count = 0;
for (int j = window[i]; j < (window[i]+size); j++) {
average += probabilityProfile[j];
count++;
}
average = average / count;
//save this windows average
averages.push_back(average);
}
return averages;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "findQav");
exit(1);
}
}
//***************************************************************************************************************
//seqs have already been masked
vector< vector<float> > DeCalculator::getQuantiles(vector<Sequence*> seqs, vector<int> windowSizesTemplate, int window, vector<float> probProfile, int increment, int start, int end) {
try {
vector< vector<float> > quan;
//percentage of mismatched pairs 1 to 100
quan.resize(100);
//for each sequence
for(int i = start; i < end; i++){
m->mothurOut("Processing sequence " + toString(i)+ "\n");
Sequence* query = new Sequence(seqs[i]->getName(), seqs[i]->getAligned());
//compare to every other sequence in template
for(int j = 0; j < i; j++){
Sequence* subject = new Sequence(seqs[j]->getName(), seqs[j]->getAligned());
if (m->getControl_pressed()) { delete query; delete subject; return quan; }
map<int, int> trim;
map<int, int>::iterator it;
trimSeqs(query, subject, trim);
it = trim.begin();
int front = it->first; int back = it->second;
//reset window for each new comparison
windowSizesTemplate[i] = window;
vector<int> win = findWindows(query, front, back, windowSizesTemplate[i], increment);
vector<float> obsi = calcObserved(query, subject, win, windowSizesTemplate[i]);
vector<float> q = findQav(win, windowSizesTemplate[i], probProfile);
float alpha = getCoef(obsi, q);
vector<float> exp = calcExpected(q, alpha);
float de = calcDE(obsi, exp);
float dist = calcDist(query, subject, front, back);
dist = ceil(dist);
quan[dist].push_back(de);
delete subject;
}
delete query;
}
return quan;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "getQuantiles");
exit(1);
}
}
//********************************************************************************************************************
//sorts lowest to highest
inline bool compareQuanMembers(quanMember left, quanMember right){
return (left.score < right.score);
}
//***************************************************************************************************************
//this was going to be used by pintail to increase the sensitivity of the chimera detection, but it wasn't quite right. may want to revisit in the future...
void DeCalculator::removeObviousOutliers(vector< vector<float> >& quantiles, int num) {
try {
for (int i = 0; i < quantiles.size(); i++) {
//find mean of this quantile score
sort(quantiles[i].begin(), quantiles[i].end());
vector<float> temp;
if (quantiles[i].size() != 0) {
float high = quantiles[i][int(quantiles[i].size() * 0.99)];
float low = quantiles[i][int(quantiles[i].size() * 0.01)];
//look at each value in quantiles to see if it is an outlier
for (int j = 0; j < quantiles[i].size(); j++) {
//is this score between 1 and 99%
if ((quantiles[i][j] > low) && (quantiles[i][j] < high)) {
temp.push_back(quantiles[i][j]);
}
}
}
quantiles[i] = temp;
}
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "removeObviousOutliers");
exit(1);
}
}
//***************************************************************************************************************
float DeCalculator::findAverage(vector<float> myVector) {
try{
float total = 0.0;
for (int i = 0; i < myVector.size(); i++) { total += myVector[i]; }
float average = total / (float) myVector.size();
return average;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "findAverage");
exit(1);
}
}
//***************************************************************************************************************
float DeCalculator::getCoef(vector<float> obs, vector<float> qav) {
try {
//find average prob for this seqs windows
float probAverage = findAverage(qav);
//find observed average
float obsAverage = findAverage(obs);
float coef = obsAverage / probAverage;
return coef;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "getCoef");
exit(1);
}
}
//***************************************************************************************************************
//gets closest matches to each end, since chimeras will most likely have different parents on each end
vector<Sequence> DeCalculator::findClosest(Sequence querySeq, vector<Sequence*>& thisTemplate, vector<Sequence*>& thisFilteredTemplate, int numWanted, int minSim) {
try {
//indexes.clear();
vector<Sequence> seqsMatches;
vector<SeqDist> distsLeft;
vector<SeqDist> distsRight;
DistCalc* distcalculator = new eachGapDist(1.0);
string queryUnAligned = querySeq.getUnaligned();
int numBases = int(queryUnAligned.length() * 0.33);
string leftQuery = ""; //first 1/3 of the sequence
string rightQuery = ""; //last 1/3 of the sequence
string queryAligned = querySeq.getAligned();
//left side
bool foundFirstBase = false;
int baseCount = 0;
int leftSpot = 0;
int firstBaseSpot = 0;
for (int i = 0; i < queryAligned.length(); i++) {
//if you are a base
if (isalpha(queryAligned[i])) {
baseCount++;
if (!foundFirstBase) { foundFirstBase = true; firstBaseSpot = i; }
}
//eliminate opening .'s
if (foundFirstBase) { leftQuery += queryAligned[i]; }
//if you have 1/3
if (baseCount >= numBases) { leftSpot = i; break; } //first 1/3
}
//right side - count through another 1/3, so you are at last third
baseCount = 0;
int rightSpot = 0;
for (int i = leftSpot; i < queryAligned.length(); i++) {
//if you are a base
if (isalpha(queryAligned[i])) { baseCount++; }
//if you have 1/3
if (baseCount > numBases + 1) { rightSpot = i; break; } //last 1/3
}
//trim end
//find last position in query that is a non gap character
int lastBaseSpot = queryAligned.length()-1;
for (int j = queryAligned.length()-1; j >= 0; j--) {
if (isalpha(queryAligned[j])) {
lastBaseSpot = j;
break;
}
}
rightQuery = queryAligned.substr(rightSpot, (lastBaseSpot-rightSpot+1)); //sequence from pos spot to end
Sequence queryLeft(querySeq.getName(), leftQuery);
Sequence queryRight(querySeq.getName(), rightQuery);
for(int j = 0; j < thisFilteredTemplate.size(); j++){
string dbAligned = thisFilteredTemplate[j]->getAligned();
string leftDB = dbAligned.substr(firstBaseSpot, (leftSpot-firstBaseSpot+1)); //first 1/3 of the sequence
string rightDB = dbAligned.substr(rightSpot, (lastBaseSpot-rightSpot+1)); //last 1/3 of the sequence
Sequence dbLeft(thisFilteredTemplate[j]->getName(), leftDB);
Sequence dbRight(thisFilteredTemplate[j]->getName(), rightDB);
double distLeft = distcalculator->calcDist(queryLeft, dbLeft);
double distRight = distcalculator->calcDist(queryRight, dbRight);
SeqDist subjectLeft;
subjectLeft.seq = nullptr;
subjectLeft.dist = distLeft;
subjectLeft.index = j;
distsLeft.push_back(subjectLeft);
SeqDist subjectRight;
subjectRight.seq = nullptr;
subjectRight.dist = distRight;
subjectRight.index = j;
distsRight.push_back(subjectRight);
}
delete distcalculator;
//sort by smallest distance
sort(distsRight.begin(), distsRight.end(), compareSeqDist);
sort(distsLeft.begin(), distsLeft.end(), compareSeqDist);
//merge results
map<string, string> seen;
map<string, string>::iterator it;
vector<SeqDist> dists;
float lastRight = distsRight[0].dist;
float lastLeft = distsLeft[0].dist;
float maxDist = 1.0 - (minSim / 100.0);
for (int i = 0; i < numWanted+1; i++) {
if (m->getControl_pressed()) { return seqsMatches; }
//add left if you havent already
it = seen.find(thisTemplate[distsLeft[i].index]->getName());
if (it == seen.end() && distsLeft[i].dist <= maxDist) {
dists.push_back(distsLeft[i]);
seen[thisTemplate[distsLeft[i].index]->getName()] = thisTemplate[distsLeft[i].index]->getName();
lastLeft = distsLeft[i].dist;
}
//add right if you havent already
it = seen.find(thisTemplate[distsRight[i].index]->getName());
if (it == seen.end() && distsRight[i].dist <= maxDist) {
dists.push_back(distsRight[i]);
seen[thisTemplate[distsRight[i].index]->getName()] = thisTemplate[distsRight[i].index]->getName();
lastRight = distsRight[i].dist;
}
if (i == numWanted) { break; }
}
//are we still above the minimum similarity cutoff
if ((lastLeft >= minSim) || (lastRight >= minSim)) {
//add in ties from left
int i = numWanted;
while (i < distsLeft.size()) {
if (distsLeft[i].dist == lastLeft) { dists.push_back(distsLeft[i]); }
else { break; }
i++;
}
//add in ties from right
i = numWanted;
while (i < distsRight.size()) {
if (distsRight[i].dist == lastRight) { dists.push_back(distsRight[i]); }
else { break; }
i++;
}
}
for (int i = 0; i < dists.size(); i++) {
if ((thisTemplate[dists[i].index]->getName() != querySeq.getName()) && (((1.0-dists[i].dist)*100) >= minSim)) {
Sequence temp(thisTemplate[dists[i].index]->getName(), thisTemplate[dists[i].index]->getAligned()); //have to make a copy so you can trim and filter without stepping on eachother.
seqsMatches.push_back(temp);
}
}
return seqsMatches;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "findClosest");
exit(1);
}
}
//***************************************************************************************************************
Sequence* DeCalculator::findClosest(Sequence* querySeq, vector<Sequence*> db) {
try {
Sequence* seqsMatch;
DistCalc* distcalculator = new eachGapDist(1.0);
int index = 0;
int smallest = 1000000;
for(int j = 0; j < db.size(); j++){
double dist = distcalculator->calcDist(*querySeq, *db[j]);
if (dist < smallest) {
smallest = dist;
index = j;
}
}
delete distcalculator;
seqsMatch = new Sequence(db[index]->getName(), db[index]->getAligned()); //have to make a copy so you can trim and filter without stepping on eachother.
return seqsMatch;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "findClosest");
exit(1);
}
}
/***************************************************************************************************************/
map<int, int> DeCalculator::trimSeqs(Sequence& query, vector<Sequence>& topMatches) {
try {
int frontPos = 0; //should contain first position in all seqs that is not a gap character
int rearPos = query.getAligned().length();
//********find first position in topMatches that is a non gap character***********//
//find first position all query seqs that is a non gap character
for (int i = 0; i < topMatches.size(); i++) {
string aligned = topMatches[i].getAligned();
int pos = 0;
//find first spot in this seq
for (int j = 0; j < aligned.length(); j++) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos > frontPos) { frontPos = pos; }
}
string aligned = query.getAligned();
int pos = 0;
//find first position in query that is a non gap character
for (int j = 0; j < aligned.length(); j++) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos > frontPos) { frontPos = pos; }
//********find last position in topMatches that is a non gap character***********//
for (int i = 0; i < topMatches.size(); i++) {
string aligned = topMatches[i].getAligned();
int pos = aligned.length();
//find first spot in this seq
for (int j = aligned.length()-1; j >= 0; j--) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos < rearPos) { rearPos = pos; }
}
aligned = query.getAligned();
pos = aligned.length();
//find last position in query that is a non gap character
for (int j = aligned.length()-1; j >= 0; j--) {
if (isalpha(aligned[j])) {
pos = j;
break;
}
}
//save this spot if it is the farthest
if (pos < rearPos) { rearPos = pos; }
map<int, int> trimmedPos;
//check to make sure that is not whole seq
if ((rearPos - frontPos - 1) <= 0) {
query.setAligned("");
//trim topMatches
for (int i = 0; i < topMatches.size(); i++) {
topMatches[i].setAligned("");
}
}else {
//trim query
string newAligned = query.getAligned();
newAligned = newAligned.substr(frontPos, (rearPos-frontPos+1));
query.setAligned(newAligned);
//trim topMatches
for (int i = 0; i < topMatches.size(); i++) {
newAligned = topMatches[i].getAligned();
newAligned = newAligned.substr(frontPos, (rearPos-frontPos+1));
topMatches[i].setAligned(newAligned);
}
for (int i = 0; i < newAligned.length(); i++) {
trimmedPos[i] = i+frontPos;
}
}
return trimmedPos;
}
catch(exception& e) {
m->errorOut(e, "DeCalculator", "trimSequences");
exit(1);
}
}
//***************************************************************************************************************
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