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#include "matepair.h"
//nextera mp adapters
string adapter1 = "CTGTCTCTTATACACATCT";
string adapter2 = "AGATGTGTATAAGAGACAG";
string adapterj = adapter1+adapter2;
//EXTERNAL adapters. this are used to clip very short dna fragments where R1 goes into R2
// string r1_external_adapter = "GTGACTGGAGTTCAGACGTGTGCTCTTCCGATC";
// string r2_external_adapter = "ACACTCTTTCCCTACACGACGCTCTTCCGATC";
string r1_external_adapter = "GATCGGAAGAGCACACGTCTGAACTCCAGTCAC";
string r2_external_adapter = "GATCGGAAGAGCGTCGTGTAGGGAAAGAGTGT";
#define DEBUG 0
#define MIN3(a, b, c) ((a) < (b) ? ((a) < (c) ? (a) : (c)) : ((b) < (c) ? (b) : (c)))
//only handles substitution errors in adapter (faster)
int hamming_match(string & target,string & query,int minoverlap,float similarity)
{
int L1=target.size();
int L2=query.size();
if(L2>=L1)
{
return(L1);
}
assert((int)L1>=minoverlap);
//check for full query matches.
int maxdist = ceil ( (1.-similarity) * L2);
int mini=L1,mind=L2;
for(int i=0;i<=(L1-L2);i++)
{
int d=0,j=0;
while(j<L2&&d<maxdist)
{
d += target[i+j]!=query[j];
j++;
}
if(d<mind)
{
mini=i;
mind=d;
}
}
if(mind<maxdist)
{
return(mini);
}
//no full match. check the edges.
mini=L1,mind=L2;
for(int i=L2-1;i>=minoverlap;i--)
{
maxdist = ceil ( (1.-similarity) * i);
//check the front of target
int j=L2-i,d=0;
while(j<L2&&d<maxdist)
{
d+=target[j-L2+i]!=query[j];
j++;
}
if(d<mind)
{
mini=i-L2;
mind=d;
}
//check the back of target
int d_back=0;
j=L1-i;
while(j<L1&&d_back<maxdist)
{
d_back+=target[j]!=query[j-L1+i];
j++;
}
if(d_back<mind)
{
mini=L1-i;
mind=d_back;
}
}
if(mind<maxdist)
{
return(mini);
}
return L1;
}
int matePair::findAdapter(string & s,int minoverlap,float similarity,bool use_hamming)
{
unsigned int L1 = s.size();
unsigned int L2 = adapter1.size();
int a;//this is the start location of the adapter
// match to entire adapter
if(use_hamming)
{
a = hamming_match(s,adapterj,minoverlap,similarity);
}
else
{
die("only hamming distance available");
}
if(a<(int)L1)
{
return a;
}
// match to first half
if(use_hamming)
{
a = hamming_match(s,adapter1,minoverlap,similarity);
}
else
{
die("only hamming distance available");
}
if(a<(int)L1)
{
return a;
}
//second half
if(use_hamming)
{
a = hamming_match(s,adapter2,minoverlap,similarity);
}
else
{
die("only hamming distance available");
}
if(a<(int)L1)
{
return(a-L2);
}
//check for shredded junction adapter (aggressive detection mode)
if(_aggressive)
{
for(int i=0;i<nseed;i++)
{
int start=L1;
if(similarity<1)
{
start=hamming_match(s,seeds[i],seedsize,similarity);
}
else
{
start = s.find(seeds[i]);
}
if(start<L1)
{//found a seed
return(start-i);
}
}
}
//ok nothing found. return end of string.
return(L1);
}
//returns min( hamming( s1[offset1,offset+L], s2[offset2,offset2+L] ) , maxd )
int hamming(string & s1,string & s2,int offset1, int offset2,int L,int maxd) {
int d = 0;
for(int i=0;i<L;i++) {
int j1=offset1+i;
int j2=offset2+i;
if(j1>=0 && j1<(int)s1.size() && j2>=0 && j2<(int)s2.size() ) {
d+=(s1[j1]!=s2[j2]);
if(d>maxd) {
d=s1.length();
break;
}
}
}
return(d);
}
int overlap(string & s1,string & s2,int minoverlap,float similarity) {
int mini=0,mind,minL;
if(s1.size()<s2.size()) minL=s1.size();
else minL=s2.size();
mind=minL;
if(minL<minoverlap)
return(0);
for(int i=minoverlap;i<minL;i++) {
int maxdist = ceil( (1. - similarity) * i );
int d = hamming(s1,s2,s1.size()-i,0,i,maxdist);
if(d<mind && d<maxdist) {
mind=d;
mini=i;
}
}
if(DEBUG>1) cerr << "mind = "<<mind<<"\tmini = "<<mini<<endl;
return(mini);
}
//checks for an overlap between r1(suffix) and r2(prefix). if there is overlap create new single read in output return 1
//if no overlap return 0
int matePair::joinReads(fqread & r1,fqread & r2,fqread & output) {
if(r1.l<minoverlap || r2.l<minoverlap || !joinreads)
return(0);
int w = overlap(r1.s,r2.s,minoverlap,similarity);
if(DEBUG>0) cerr << "w = "<<w<<endl;
if(w==0)
return(0);
else {
output.h=r1.h;
output.l3=r1.l3;
output.s = r1.s + r2.s.substr(w);
output.q = r1.q + r2.q.substr(w);
int offset = r1.s.size() - w;
for(int i=0;i<w;i++) {//takes highest quality base.
int j = offset+w;
if( (uint8_t)r1.q[j] < (uint8_t)r2.q[i] ) {
output.s[j] = r2.s[i];
output.q[j] = r2.q[i];
}
}
output.l = output.s.size();
return(1);
}
}
int matePair::resolve_overhang(fqread & r1, fqread & r2,int a,int b) {
if(DEBUG>2)
{
cerr << "Resolving overhang"<<endl;
}
fqread tmp1 = r1.window(b,r1.l);
fqread tmp2 = r1.window(b,r1.l).rc();
if(DEBUG>2)
{
cerr << r2.s <<endl;;
cerr << tmp2.s << endl;
}
if(a<minlen)
{//preceeding dna is too small.
//TODO:could possibly merge for a big read here
if(_justmp)
{
mp.r1 = r1.mask();
mp.r2 = r2;
}
else
{
pe.r1 = tmp1;
pe.r2 = r2;
}
}
else if(joinReads(r2,tmp2,mp.r2))
{
mp.r1=r1.window(0,a);
}
else if(r1.notN(b,r1.l)>r1.notN(0,a) && !preserve_mp)
{
pe.r1=tmp1;
pe.r2=r2;
if(a>=minlen)
se = r1.window(0,a);
}
else
{
if(tmp1.l>=minlen)
se = tmp1;
mp.r1=r1.window(0,a);
mp.r2=r2;
}
return(0);
}
matePair::matePair()
{
_aggressive=false;
//build seeds;
seedsize=adapter1.length();
nseed=0;
for(int i=0;i<adapterj.length()-seedsize;i++)
{
seeds.push_back(adapterj.substr(i,seedsize));
nseed++;
}
}
matePair::~matePair()
{
}
int matePair::clear()
{
mp.r1.clear();
mp.r2.clear();
pe.r1.clear();
pe.r2.clear();
unknown.r1.clear();
unknown.r2.clear();
se.clear();
return(0);
}
//this is just a simple routine to trim edges off matepairs where the Nx adapter was not detected
//removes perfect adapter matches on edges that are < minoverlap
int matePair::trimUnknown()
{
//start
int a1=0,a2=0,b1=unknown.r1.l,b2=unknown.r2.l;
for(int i=3;i<=minoverlap;i++)
{
int offset = unknown.r1.l-i;
int maxd = ceil ( (1.-similarity) * i);
if(hamming(unknown.r1.s,adapter1,offset,0,i,maxd)<maxd)
a1=offset;
offset = unknown.r2.l-i;
if(hamming(unknown.r2.s,adapter1,offset,0,i,maxd)<maxd)
a2=offset;
}
if(DEBUG>0)
if(a1>0||a2>0)
cerr << "trimUnknown: " << a1 << " " << b1 << " " << a2 << " " << b2 << endl;
if(a1>0)
unknown.r1 = unknown.r1.window(0,a1);
if(a2>0)
unknown.r2 = unknown.r2.window(0,a2);
return(0);
}
//gets rid of the rare case where external adapters are present (isize < 2*L)
bool matePair::trimExternal(readPair& rp)
{
bool found = false;
int a,b;
unsigned int tmp = rp.r1.s.find(r1_external_adapter);//PERFECT MATCH?
if(tmp>=rp.r1.s.size()) //PARTIAL MATCH?
a = hamming_match(rp.r1.s,r1_external_adapter,minoverlap,similarity);
else a = (int)tmp;
tmp = rp.r2.s.find(r2_external_adapter);//PERFECT MATCH?
if(tmp>=rp.r1.s.size()) //PARTIAL MATCH?
b = hamming_match(rp.r2.s,r2_external_adapter,minoverlap,similarity);
else
b = (int)tmp;
if(DEBUG>1) {
if((a>0 && a<rp.r1.l)||(b>0 && b<rp.r2.l)) {
cerr << "EXTERNAL ADAPTER DETECTED " << a << " " << b << endl;
if(a>0 && a<rp.r1.l) {
rp.r1.window(a,rp.r1.l).print();
}
if(b>0 && b<rp.r2.l) {
rp.r2.window(b,rp.r2.l).print();
}
// rp.r1.print();
// rp.r2.print();
}
}
// OK NO ADAPTERS FOUND, LETS TRY LOOKING FOR AN OVERLAP -> PAIRED END FRAG
if(!(a>0 && a<rp.r1.l)&&!(b>0 && b<rp.r2.l)) {
fqread rc2 = rp.r2.rc();
int mini=rp.r1.l,mind=rp.r1.l;
for(int i=0;i<(rp.r1.l-minlen);i++)
{
int compare_length = rp.r2.l-i;
int maxdist = ceil ( (1.-similarity) * compare_length);
int d = hamming(rp.r1.s,rc2.s,0,i,rp.r2.l-i,maxdist);
if(d<mind&&d<maxdist)
{
mini = i;
mind = d;
}
}
if(mini<rp.r1.l)
{
a=rp.r1.l-mini;
b=rp.r2.l-mini;
if(DEBUG>1) cerr <<"OVERLAP FOUND "<< a <<" " << b<<endl;
}
}
if((a>0 && a<rp.r1.l)||(b>0 && b<rp.r2.l))
{
found = true;
if(_justmp)
{
mp.r1 = rp.r1;
mp.r2 = rp.r2;
if(a<rp.r1.l)
mp.r1 = rp.r1.mask();
if(b<rp.r2.l)
mp.r2 = rp.r2.mask();
}
else
{
if(a<rp.r1.l)
pe.r1 = rp.r1.window(0,a);
else
pe.r1 = rp.r1;
if(b<rp.r2.l)
pe.r2 = rp.r2.window(0,b);
else
pe.r2 = rp.r2;
}
}
return(found);
}
//aligns s2 to s1 with sim>=sim. returns s1.size() if no alignment found
unsigned int matePair::ham_align(string & s1,string & s2)
{
if(s1.size()<s2.size())
return(s1.size());
int L1 = s1.size();
int L2 = s2.size();
assert(L2>=minoverlap);
int maxd = ceil ( (1.-similarity) * L2);
int mind=maxd,mini=L1;
int d;
for(int i=0;i<(L1-L2);i++)
{
d = hamming(s1,s2,L1-i-L2,0,L2,maxd);
if(d<mind)
{
mind=d;
mini=L1-i;
}
}
if(d>maxd)//hit wasnt good enough
mini=L1;
return(mini);
}
//checks the right end of a read for partial adapter hit
int checkRight(string & s1,string & adapter,int offset,int minoverlap,float similarity)
{
assert(offset <= (s1.size()-minoverlap));
int a=s1.size();
int mind = s1.size();
for(int i=offset;i<(s1.size()-minoverlap);i++)
{
int compare_len = (s1.size() - i);
int maxdist = ceil(compare_len * (1. - similarity));
int d = hamming(s1,adapter,i,0,compare_len,maxdist);
if(d<mind&&d<maxdist)
{
a=i;
mind=d;
}
}
return(a);
}
//int matePair::build(readPair & readpair,int minoverlap,float similarity,int minlen,bool joinreads,bool use_hamming) {
int matePair::build(readPair& readpair,int minovl,float sim,int ml,bool jr,bool uh,bool pmp,bool jmp)
{
clear();
_justmp=jmp;
preserve_mp=pmp;
minoverlap=minovl;
similarity=sim;
minlen=ml;
joinreads=jr;
use_hamming=uh;
int L1 = readpair.r1.l;
int L2 = readpair.r2.l;
if(L1<minoverlap||L2<minoverlap)
{
cerr << "WARNING: read with length < minimum overlap length ("<<minoverlap<<"). Will discard this read pair:"<<endl;
cerr << "@" << readpair.r1.h <<endl;
cerr << readpair.r1.s <<endl;
cerr << readpair.r1.l3 <<endl;
cerr << readpair.r1.q <<endl;
cerr << "@" << readpair.r2.h <<endl;
cerr << readpair.r2.s <<endl;
cerr << readpair.r2.l3 <<endl;
cerr << readpair.r2.q <<endl;
return(0);
}
int a1 = findAdapter(readpair.r1.s, minoverlap, similarity,use_hamming);
int a2 = findAdapter(readpair.r2.s, minoverlap, similarity,use_hamming);
int b1 = a1+adapterj.size();
int b2 = a2+adapterj.size();
if(DEBUG>1)
{
cerr << "read L1 = "<<L1<<endl;
cerr << "read L2 = "<<L2<<endl;
cerr << "adapter locations (first pass): "<<a1 << " " << b1 << " " << a2 << " " << b2 << endl;
}
//check for extra unexpected adapter copies (entire read pair is discarded in this case)
if(a1<L1)
{
fqread tmp = readpair.r1.mask(max(0,a1),min(b1,L1));
if(findAdapter(tmp.s, minoverlap, similarity,use_hamming) < L1)
{
return(1);
}
}
if(a2<L2)
{
fqread tmp = readpair.r2.mask(max(0,a2),min(b2,L2));
if(findAdapter(tmp.s, minoverlap, similarity,use_hamming) < L2)
{
return(1);
}
}
fqread rc1 = readpair.r1.rc();
fqread rc2 = readpair.r2.rc();
if(a1==L1&&b2<(L2-minoverlap))
{//try to overlay the r2 overhang to r1 -> finds adapter on r1
string overhang = rc2.s.substr(0,rc2.l-b2);
a1 = ham_align(readpair.r1.s,overhang);
b1 = a1+adapterj.size();
}
if(a2==L2&&b1<(L1-minoverlap))
{//vice-versa
string overhang = rc1.s.substr(0,rc1.l-b1);
a2 = ham_align(readpair.r2.s,overhang);
b2 = a2+adapterj.size();
}
if(DEBUG>1)
{
cerr << "adapter locations (second pass): "<<a1 << " " << b1 << " " << a2 << " " << b2 << endl;
}
int minoverlap2 = 1; //final attempt to find unidentified adapters
if(a1<L1&&a2==L2)//we know R1 has adapter. try check R2 for adapter with more liberal thresholds
a2 = checkRight(readpair.r2.s,adapter1, L2-minoverlap, minoverlap2, similarity);
if(a2<L2&&a1==L1)//vice-versa
a1 = checkRight(readpair.r1.s,adapter1, L1-minoverlap, minoverlap2, similarity);
if(DEBUG>1)
{
cerr << "adapter locations (third pass): "<<a1 << " " << b1 << " " << a2 << " " << b2 << endl;
}
if(a1==L1 && a2==L2) {//no adapter found
// we could potentially run if(!joinReads(readpair.r1,rc2,se)) but this tends to give a lot of false joins
// possible improvement: check for r1/r2 overlap in absence of adapter -> overlap implies PE
if(!trimExternal(readpair)) {
unknown=readPair(readpair.r1,readpair.r2);
trimUnknown();
}
if(DEBUG>1) {
cerr << "CASE A"<<endl;
cerr<<"UNKNOWN: "<<endl;
unknown.print();
cerr<<"PE: "<<endl;
pe.print();
}
}
else {//adapter found.
bool both_have_adapter = a1<L1 && a2<L2;
bool R1_has_adapter_at_end = a1<L1 && b1>=(L1-minlen);
bool R2_has_adapter_at_end = a2<L2 && b2>=(L2-minlen);
if(a1<minlen&&a2<minlen)
{//very short template. discard
return(0);
}
else if(a1<(L1-minoverlap) && a2<minlen)
{//r2 redundant
if(_justmp)
mp=readPair(readpair.r1.window(0,a1),readpair.r2.mask()) ;
else
se = readpair.r1.window(0,a1);
if(DEBUG>1) cerr << "CASE B"<<endl;
}
else if(a2<(L2-minoverlap) && a1<minlen)
{//r1 redundant
if(_justmp)
mp=readPair(readpair.r1.mask(),readpair.r2.window(0,a2));
else
se = readpair.r2.window(0,a2);
if(DEBUG>1) cerr << "CASE C"<<endl;
}
else if(a1>=(L1-minoverlap) && a2<minlen)
{//obvious PE
if(a1>=minlen && (L2-b2)>=minlen)
{
if(_justmp)
{
mp=readPair(readpair.r1.window(0,a1),readpair.r2.mask()) ;
}
else
{
pe.r1 = readpair.r1.window(0,a1);
pe.r2 = readpair.r2.window(b2,b2+a1);
}
}
if(DEBUG>1) cerr << "CASE D"<<endl;
}
else if(a2>=(L2-minoverlap) && a1<minlen)
{//obvious PE
if(a2>=minlen && (L1-b1)>=minlen)
{
if(_justmp)
{
mp=readPair(readpair.r1.mask(),readpair.r2.window(0,a2));
}
else
{
pe.r1 = readpair.r1.window(b1,b1+a2);
pe.r2 = readpair.r2.window(0,a2);
}
}
if(DEBUG>1) cerr << "CASE E"<<endl;
}
else if(both_have_adapter||R1_has_adapter_at_end||R2_has_adapter_at_end)
{
//standard mp
mp.r1=readpair.r1.window(0,a1);
mp.r2=readpair.r2.window(0,a2);
if(DEBUG>1) cerr << "CASE F"<<endl;
/*
if((L1-b1)>minlen && b1<=b2)
se = readpair.r1.window(b1);
if((L2-b2)>minlen && b2<b1)
se = readpair.r2.window(b2);
*/
}
else if(b1<L1 && a2==L2)
{
resolve_overhang(readpair.r1,readpair.r2,a1,b1);
if(DEBUG>1) cerr << "CASE G"<<endl;
}
else if(b2<L2 && a1==L1)
{
resolve_overhang(readpair.r2,readpair.r1,a2,b2);
fqread swap1 = pe.r1;
pe.r1 = pe.r2;
pe.r2 = swap1;
fqread swap2 = mp.r1;
mp.r1 = mp.r2;
mp.r2 = swap2;
if(DEBUG>1) cerr << "CASE H"<<endl;
}
}
return(0);
}
nxtrimWriter::nxtrimWriter(string prefix,bool jmp,bool separate) {
open(prefix,jmp,separate);
}
int nxtrimWriter::open(string prefix,bool jmp,bool separate) {
if(prefix=="-")
die("bad output file name: "+prefix);
n_mp=0;
n_pe=0;
n_se=0;
n_unk=0;
_justmp = jmp;
_write_un=_write_mp=_write_se=_write_pe=true;
if(separate) {
mp_out.open(prefix+"_R1.mp.fastq.gz", prefix+"_R2.mp.fastq.gz");
unknown_out.open(prefix+"_R1.unknown.fastq.gz",prefix+"_R2.unknown.fastq.gz");
}
else {
mp_out.open(prefix+".mp.fastq.gz");
unknown_out.open(prefix+".unknown.fastq.gz");
}
if(!_justmp) {
if(separate) pe_out.open(prefix+"_R1.pe.fastq.gz",prefix+"_R2.pe.fastq.gz");
else pe_out.open(prefix+".pe.fastq.gz");
se_out.open(prefix+".se.fastq.gz");
}
else {
_write_se=_write_pe=false;
}
return(0);
}
nxtrimWriter::nxtrimWriter() {}
int nxtrimWriter::open()
{
n_mp=0;
n_pe=0;
n_se=0;
n_unk=0;
_justmp = true;
_write_un=_write_mp=true;
_write_se=_write_pe=false;
mp_out.open("-");
unknown_out.open("-");
return(0);
}
int nxtrimWriter::write(matePair & m) {
if(_write_mp) n_mp+=mp_out.write(m.mp);
if(_write_un) n_unk+=unknown_out.write(m.unknown);
if(_write_pe) n_pe+=pe_out.write(m.pe);
if(_write_se) n_se+=se_out.write(m.se);
if(DEBUG>0)
cerr << "Wrote: n_mp="<<n_mp<<" n_unk="<<n_unk<<" n_pe="<<n_pe<<" n_se="<<n_se<<endl;
return(0);
}
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