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#include <cmath>
#include "Genome.h"
#include "IncludeDefine.h"
#include "Parameters.h"
#include "SuffixArrayFuns.h"
#include "PackedArray.h"
#include "TimeFunctions.h"
#include "ErrorWarning.h"
#include "GTF.h"
#include "SjdbClass.h"
#include "sjdbLoadFromFiles.h"
#include "sjdbPrepare.h"
#include "genomeParametersWrite.h"
#include "sjdbInsertJunctions.h"
#include "genomeScanFastaFiles.h"
#include "genomeSAindex.h"
#include "serviceFuns.cpp"
#include "streamFuns.h"
#include "SequenceFuns.h"
char* globalG;
uint globalL;
inline int funCompareSuffixes ( const void *a, const void *b){
uint *ga=(uint*)((globalG-7LLU)+(*((uint*)a)));
uint *gb=(uint*)((globalG-7LLU)+(*((uint*)b)));
uint jj=0;
int ii=0;
uint va=0,vb=0;
uint8 *va1, *vb1;
while (jj < globalL) {
va=*(ga-jj);
vb=*(gb-jj);
#define has5(v) ((((v)^0x0505050505050505) - 0x0101010101010101) & ~((v)^0x0505050505050505) & 0x8080808080808080)
if (has5(va) && has5(vb))
{//there is 5 in the sequence - only compare bytes before 5
va1=(uint8*) &va;
vb1=(uint8*) &vb;
for (ii=7;ii>=0;ii--)
{
if (va1[ii]>vb1[ii])
{
return 1;
} else if (va1[ii]<vb1[ii])
{
return -1;
} else if (va1[ii]==5)
{//va=vb at the end of chr
if ( *((uint*)a) > *((uint*)b) )
{//anti-stable order,since indexes are sorted in the reverse order
return -1;
} else
{//a cannot be equal to b
return 1;
};
};
};
} else
{//no 5, simple comparison
if (va>vb)
{
return 1;
} else if (va<vb)
{
return -1;
};
};
jj++;
};
//suffixes are equal up to globalL, no simply compare the indexes
if ( *((uint*)a) > *((uint*)b) )
{//anti-stable order,since indexes are sorted in the reverse order
return -1;
} else
{//a cannot be equal to b
return 1;
};
};
inline uint funG2strLocus (uint SAstr, uint const N, char const GstrandBit, uint const GstrandMask) {
bool strandG = (SAstr>>GstrandBit) == 0;
SAstr &= GstrandMask;
if ( !strandG ) SAstr += N;
return SAstr;
};
void Genome::genomeGenerate() {
//check parameters
createDirectory(pGe.gDir, P.runDirPerm, "--genomeDir", P);
{//move Log.out file into genome directory
string logfn=pGe.gDir+"Log.out";
if ( rename( P.outLogFileName.c_str(), logfn.c_str() ) ) {
warningMessage("Could not move Log.out file from " + P.outLogFileName + " into " + logfn + ". Will keep " + P.outLogFileName +"\n", \
std::cerr, P.inOut->logMain, P);
} else {
P.outLogFileName=logfn;
};
};
if (sjdbOverhang<=0 && (pGe.sjdbFileChrStartEnd.at(0)!="-" || pGe.sjdbGTFfile!="-")) {
ostringstream errOut;
errOut << "EXITING because of FATAL INPUT PARAMETER ERROR: for generating genome with annotations (--sjdbFileChrStartEnd or --sjdbGTFfile options)\n";
errOut << "you need to specify >0 --sjdbOverhang\n";
errOut << "SOLUTION: re-run genome generation specifying non-zero --sjdbOverhang, which ideally should be equal to OneMateLength-1, or could be chosen generically as ~100\n";
exitWithError(errOut.str(),std::cerr, P.inOut->logMain, EXIT_CODE_INPUT_FILES, P);
};
if (pGe.sjdbFileChrStartEnd.at(0)=="-" && pGe.sjdbGTFfile=="-") {
if (P.parArray.at(P.pGe.sjdbOverhang_par)->inputLevel>0 && sjdbOverhang>0) {
ostringstream errOut;
errOut << "EXITING because of FATAL INPUT PARAMETER ERROR: when generating genome without annotations (--sjdbFileChrStartEnd or --sjdbGTFfile options)\n";
errOut << "do not specify >0 --sjdbOverhang\n";
errOut << "SOLUTION: re-run genome generation without --sjdbOverhang option\n";
exitWithError(errOut.str(),std::cerr, P.inOut->logMain, EXIT_CODE_INPUT_FILES, P);
};
sjdbOverhang=0;
};
//time
time_t rawTime;
string timeString;
time(&rawTime);
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... starting to generate Genome files\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... starting to generate Genome files\n" <<flush;
//define some parameters from input parameters
genomeChrBinNbases=1LLU << pGe.gChrBinNbits;
nGenome = genomeScanFastaFiles(P,NULL,false,*this);//first scan the fasta file to find all the sizes
genomeSequenceAllocate(nGenome, nG1alloc, G, G1);
genomeScanFastaFiles(P,G,true,*this); //load the genome sequence
uint64 nGenomeTrue=0;
for (auto &cl : chrLength)
nGenomeTrue += cl; //nGenomeTrue = sum of chr lengths
P.inOut->logMain <<"Genome sequence total length = " << nGenomeTrue << "\n";
P.inOut->logMain <<"Genome size with padding = "<< nGenome <<"\n";
//consensusSequence(); //replace with consensus allele DEPRECATED
SjdbClass sjdbLoci; //will be filled in transcriptGeneSJ below
GTF mainGTF(*this, P, pGe.gDir, sjdbLoci); //this loads exonLoci and gene/transcript metadata only, sjdbLoci is not filled
Genome::transformGenome(&mainGTF);
mainGTF.superTranscript(); //this may change the genome into (Super)Transcriptome
chrBinFill();//chrBin is first used in the transcriptGeneSJ below
mainGTF.transcriptGeneSJ(pGe.gDir);
sjdbLoadFromFiles(P,sjdbLoci);//this will not be transformed. TODO prevent this parameter combination
if (pGe.gSAindexNbases > log2(nGenomeTrue)/2-1) {
ostringstream warnOut;
warnOut << "--genomeSAindexNbases " << pGe.gSAindexNbases << " is too large for the genome size=" << nGenomeTrue;
warnOut << ", which may cause seg-fault at the mapping step. Re-run genome generation with recommended --genomeSAindexNbases " << int(log2(nGenomeTrue)/2-1);
warningMessage(warnOut.str(),P.inOut->logMain,std::cerr,P);
};
//output genome metadata
writeChrInfo(pGe.gDir);
//preparing to generate SA
for (uint ii=0;ii<nGenome;ii++) {//- strand
//if (G[ii]>5)
// cerr << ii <<" "<< G[ii]<<"\n";
G[2*nGenome-1-ii]=G[ii]<4 ? 3-G[ii] : G[ii];
};
nSA=0;
for (uint ii=0;ii<2*nGenome;ii+=pGe.gSAsparseD) {
if (G[ii]<4) {
nSA++;
};
};
// GstrandBit
GstrandBit = (char) (uint) floor(log(nGenome+P.limitSjdbInsertNsj*sjdbLength)/log(2))+1; //GstrandBit uses P.limitSjdbInsertNsj even if no insertion requested, in case it will be requested at the mapping stage
if (GstrandBit<32) GstrandBit=32; //TODO: should not this be 31? Need to test for small genomes. TODO: use simple access function for SA
P.inOut->logMain <<"Estimated genome size with padding and SJs: total=genome+SJ="<<nGenome+P.limitSjdbInsertNsj*sjdbLength<<" = "<<nGenome<<" + "<<P.limitSjdbInsertNsj*sjdbLength<<"\n";
P.inOut->logMain << "GstrandBit=" << int(GstrandBit) <<"\n";
GstrandMask = ~(1LLU<<GstrandBit);
SA.defineBits(GstrandBit+1,nSA);
if (P.sjdbInsert.yes) {//reserve space for junction insertion
SApass1.defineBits( GstrandBit+1, nSA+2*sjdbLength*min((uint64)sjdbLoci.chr.size(),P.limitSjdbInsertNsj) );//TODO: this allocation is wasteful, get a better estimate of the number of junctions
} else {//same as SA
SApass1.defineBits(GstrandBit+1,nSA);
};
P.inOut->logMain << "Number of SA indices: "<< nSA << "\n"<<flush;
//sort SA
time ( &rawTime );
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... starting to sort Suffix Array. This may take a long time...\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... starting to sort Suffix Array. This may take a long time...\n" <<flush;
// if (false)
{//sort SA chunks
for (uint ii=0;ii<nGenome;ii++) {//re-fill the array backwards for sorting
swap(G[2*nGenome-1-ii],G[ii]);
};
globalG=G;
globalL=pGe.gSuffixLengthMax/sizeof(uint);
//count the number of indices with 4nt prefix
uint indPrefN=1LLU << 16;
uint* indPrefCount = new uint [indPrefN];
memset(indPrefCount,0,indPrefN*sizeof(indPrefCount[0]));
nSA=0;
for (uint ii=0;ii<2*nGenome;ii+=pGe.gSAsparseD) {
if (G[ii]<4) {
uint p1=(G[ii]<<12) + (G[ii-1]<<8) + (G[ii-2]<<4) + G[ii-3];
indPrefCount[p1]++;
nSA++;
};
};
uint saChunkSize=(P.limitGenomeGenerateRAM-nG1alloc)/8/P.runThreadN; //number of SA indexes per chunk
saChunkSize=saChunkSize*6/10; //allow extra space for qsort
//uint saChunkN=((nSA/saChunkSize+1)/P.runThreadN+1)*P.runThreadN;//ensure saChunkN is divisible by P.runThreadN
//saChunkSize=nSA/saChunkN+100000;//final chunk size
if (P.runThreadN>1) saChunkSize=min(saChunkSize,nSA/(P.runThreadN-1));
uint saChunkN = nSA / saChunkSize + 1;//estimate
uint* indPrefStart = new uint [saChunkN*2]; //start and stop, *2 just in case
uint* indPrefChunkCount = new uint [saChunkN*2];
indPrefStart[0]=0;
saChunkN=0;//start counting chunks
uint chunkSize1=indPrefCount[0];
for (uint ii=1; ii<indPrefN; ii++) {
chunkSize1 += indPrefCount[ii];
if (chunkSize1 > saChunkSize) {
saChunkN++;
indPrefStart[saChunkN]=ii;
indPrefChunkCount[saChunkN-1]=chunkSize1-indPrefCount[ii];
chunkSize1=indPrefCount[ii];
};
};
saChunkN++;
indPrefStart[saChunkN]=indPrefN+1;
indPrefChunkCount[saChunkN-1]=chunkSize1;
P.inOut->logMain << "Number of chunks: " << saChunkN <<"; chunks size limit: " << saChunkSize*8 <<" bytes\n" <<flush;
time ( &rawTime );
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... sorting Suffix Array chunks and saving them to disk...\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... sorting Suffix Array chunks and saving them to disk...\n" <<flush;
#pragma omp parallel for num_threads(P.runThreadN) ordered schedule(dynamic,1)
for (int iChunk=0; iChunk < (int) saChunkN; iChunk++) {//start the chunk cycle: sort each chunk with qsort and write to a file
uint* saChunk=new uint [indPrefChunkCount[iChunk]];//allocate local array for each chunk
for (uint ii=0,jj=0;ii<2*nGenome;ii+=pGe.gSAsparseD) {//fill the chunk with SA indices
if (G[ii]<4) {
uint p1=(G[ii]<<12) + (G[ii-1]<<8) + (G[ii-2]<<4) + G[ii-3];
if (p1>=indPrefStart[iChunk] && p1<indPrefStart[iChunk+1]) {
saChunk[jj]=ii;
jj++;
};
//TODO: if (jj==indPrefChunkCount[iChunk]) break;
};
};
//sort the chunk
qsort(saChunk,indPrefChunkCount[iChunk],sizeof(saChunk[0]),funCompareSuffixes);
for (uint ii=0;ii<indPrefChunkCount[iChunk];ii++) {
saChunk[ii]=2*nGenome-1-saChunk[ii];
};
//write files
string chunkFileName=pGe.gDir+"/SA_"+to_string( (uint) iChunk);
ofstream & saChunkFile = ofstrOpen(chunkFileName,ERROR_OUT, P);
fstreamWriteBig(saChunkFile, (char*) saChunk, sizeof(saChunk[0])*indPrefChunkCount[iChunk],chunkFileName,ERROR_OUT,P);
saChunkFile.close();
delete [] saChunk;
saChunk=NULL;
};
time ( &rawTime );
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... loading chunks from disk, packing SA...\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... loading chunks from disk, packing SA...\n" <<flush;
//read chunks and pack into full SA
SApass1.allocateArray();
SA.pointArray(SApass1.charArray + SApass1.lengthByte-SA.lengthByte); //SA is shifted to have space for junction insertion
uint N2bit= 1LLU << GstrandBit;
uint packedInd=0;
#define SA_CHUNK_BLOCK_SIZE 10000000
uint* saIn=new uint[SA_CHUNK_BLOCK_SIZE]; //TODO make adjustable
#ifdef genenomeGenerate_SA_textOutput
ofstream SAtxtStream ((pGe.gDir + "/SAtxt").c_str());
#endif
for (uint iChunk=0;iChunk<saChunkN;iChunk++) {//load files one by one and convert to packed
ostringstream saChunkFileNameStream("");
saChunkFileNameStream<< pGe.gDir << "/SA_" << iChunk;
ifstream saChunkFile(saChunkFileNameStream.str().c_str());
while (! saChunkFile.eof()) {//read blocks from each file
uint chunkBytesN=fstreamReadBig(saChunkFile,(char*) saIn,SA_CHUNK_BLOCK_SIZE*sizeof(saIn[0]));
for (uint ii=0;ii<chunkBytesN/sizeof(saIn[0]);ii++) {
SA.writePacked( packedInd+ii, (saIn[ii]<nGenome) ? saIn[ii] : ( (saIn[ii]-nGenome) | N2bit ) );
#ifdef genenomeGenerate_SA_textOutput
SAtxtStream << saIn[ii] << "\n";
#endif
};
packedInd += chunkBytesN/sizeof(saIn[0]);
};
saChunkFile.close();
remove(saChunkFileNameStream.str().c_str());//remove the chunk file
};
#ifdef genenomeGenerate_SA_textOutput
SAtxtStream.close();
#endif
delete [] saIn;
if (packedInd != nSA ) {//
ostringstream errOut;
errOut << "EXITING because of FATAL problem while generating the suffix array\n";
errOut << "The number of indices read from chunks = "<<packedInd<<" is not equal to expected nSA="<<nSA<<"\n";
errOut << "SOLUTION: try to re-run suffix array generation, if it still does not work, report this problem to the author\n"<<flush;
exitWithError(errOut.str(),std::cerr, P.inOut->logMain, EXIT_CODE_INPUT_FILES, P);
};
//DONE with suffix array generation
for (uint ii=0;ii<nGenome;ii++) {//return to normal order for future use
swap(G[2*nGenome-1-ii],G[ii]);
};
delete [] indPrefCount;
delete [] indPrefStart;
delete [] indPrefChunkCount;
};
time ( &rawTime );
timeString=asctime(localtime ( &rawTime ));
timeString.erase(timeString.end()-1,timeString.end());
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... finished generating suffix array\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... finished generating suffix array\n" <<flush;
genomeSAindex(G, SA, P, SAi, *this);
sjdbN=0;
if (P.sjdbInsert.yes) {//insert junctions
P.sjdbInsert.outDir=pGe.gDir;
P.twoPass.pass2=false;
Genome genome1(*this); //create copy here, *this will be changed by sjdbInsertJunctions
sjdbInsertJunctions(P, *this, genome1, sjdbLoci);
};
pGe.gFileSizes.clear();
pGe.gFileSizes.push_back(nGenome);
pGe.gFileSizes.push_back(SA.lengthByte);
//write genome parameters file
genomeParametersWrite(pGe.gDir+("/genomeParameters.txt"), P, ERROR_OUT, *this);
//write genome to disk
time ( &rawTime );
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... writing Genome to disk ...\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... writing Genome to disk ...\n" <<flush;
writeGenomeSequence(pGe.gDir);
//write SA
time ( &rawTime );
P.inOut->logMain << "SA size in bytes: "<<SA.lengthByte << "\n"<<flush;
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... writing Suffix Array to disk ...\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... writing Suffix Array to disk ...\n" <<flush;
ofstream & SAout = ofstrOpen(pGe.gDir+"/SA",ERROR_OUT, P);
fstreamWriteBig(SAout,(char*) SA.charArray, (streamsize) SA.lengthByte,pGe.gDir+"/SA",ERROR_OUT,P);
SAout.close();
//write SAi
time(&rawTime);
P.inOut->logMain << timeMonthDayTime(rawTime) <<" ... writing SAindex to disk\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) <<" ... writing SAindex to disk\n" <<flush;
//write SAi to disk
ofstream & SAiOut = ofstrOpen(pGe.gDir+"/SAindex",ERROR_OUT, P);
fstreamWriteBig(SAiOut, (char*) &pGe.gSAindexNbases, sizeof(pGe.gSAindexNbases),pGe.gDir+"/SAindex",ERROR_OUT,P);
fstreamWriteBig(SAiOut, (char*) genomeSAindexStart, sizeof(genomeSAindexStart[0])*(pGe.gSAindexNbases+1),pGe.gDir+"/SAindex",ERROR_OUT,P);
fstreamWriteBig(SAiOut, SAi.charArray, SAi.lengthByte,pGe.gDir+"/SAindex",ERROR_OUT,P);
SAiOut.close();
SApass1.deallocateArray();
time(&rawTime);
timeString=asctime(localtime ( &rawTime ));
timeString.erase(timeString.end()-1,timeString.end());
time(&rawTime);
P.inOut->logMain << timeMonthDayTime(rawTime) << " ..... finished successfully\n" <<flush;
*P.inOut->logStdOut << timeMonthDayTime(rawTime) << " ..... finished successfully\n" <<flush;
};
void Genome::writeChrInfo(const string dirOut)
{//write chr information
ofstream & chrN = ofstrOpen(dirOut+"/chrName.txt",ERROR_OUT, P);
ofstream & chrS = ofstrOpen(dirOut+"/chrStart.txt",ERROR_OUT, P);
ofstream & chrL = ofstrOpen(dirOut+"/chrLength.txt",ERROR_OUT, P);
ofstream & chrNL = ofstrOpen(dirOut+"/chrNameLength.txt",ERROR_OUT, P);
for (uint ii=0;ii<nChrReal;ii++) {//output names, starts, lengths
chrN<<chrName[ii]<<"\n";
chrS<<chrStart[ii]<<"\n";
chrL<<chrLength.at(ii)<<"\n";
chrNL<<chrName[ii]<<"\t"<<chrLength.at(ii)<<"\n";
};
chrS<<chrStart[nChrReal]<<"\n";//size of the genome
chrN.close();chrL.close();chrS.close(); chrNL.close();
};
void Genome::writeGenomeSequence(const string dirOut)
{//write genome sequence
ofstream &genomeOut = ofstrOpen(dirOut+"/Genome",ERROR_OUT, P);
fstreamWriteBig(genomeOut,G,nGenome,dirOut+"/Genome",ERROR_OUT,P);
genomeOut.close();
};
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