1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165 166 167 168 169 170 171 172 173 174 175 176 177 178 179 180 181 182 183 184 185 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 202 203 204 205 206 207 208 209 210 211 212 213 214 215 216 217 218 219 220 221 222 223 224 225 226 227 228 229 230 231 232 233 234 235 236 237 238 239 240 241 242 243 244 245 246 247 248 249 250 251 252 253 254 255 256 257 258
|
#include <sys/types.h>
#include <sys/stat.h>
#include "IncludeDefine.h"
#include "Parameters.h"
#include "SequenceFuns.h"
#include "Genome.h"
#include "Chain.h"
#include "ReadAlignChunk.h"
#include "ReadAlign.h"
#include "Stats.h"
#include "genomeGenerate.h"
#include "outputSJ.h"
#include "ThreadControl.h"
#include "GlobalVariables.h"
#include "TimeFunctions.h"
#include "ErrorWarning.h"
#include "sysRemoveDir.h"
#include "BAMfunctions.h"
#include "bamSortByCoordinate.h"
#include "Transcriptome.h"
#include "signalFromBAM.h"
#include "mapThreadsSpawn.h"
#include "SjdbClass.h"
#include "sjdbInsertJunctions.h"
#include "Variation.h"
#include "Solo.h"
#include "samHeaders.h"
#include "twoPassRunPass1.h"
#include <htslib/sam.h>
#include "parametersDefault.xxd"
void usage(int usageType) {
cout << "Usage: STAR [options]... --genomeDir /path/to/genome/index/ --readFilesIn R1.fq R2.fq\n";
cout <<"Spliced Transcripts Alignment to a Reference (c) Alexander Dobin, 2009-2020\n\n";
cout << "STAR version=" << STAR_VERSION << "\n";
cout << "STAR compilation time,server,dir=" << COMPILATION_TIME_PLACE << "\n";
cout << "For more details see:\n";
cout << "<https://github.com/alexdobin/STAR>\n";
cout << "<https://github.com/alexdobin/STAR/blob/master/doc/STARmanual.pdf>\n";
if (usageType==0) {//brief
cout << "\nTo list all parameters, run STAR --help\n";
} else if (usageType==1) {//full
cout.write(reinterpret_cast<char *> (parametersDefault),
parametersDefault_len);
};
exit(0);
};
int main(int argInN, char* argIn[]) {
// If no argument is given, or the first argument is either '-h' or '--help', run usage()
if (argInN == 1) {
usage(0);
} else if (argInN == 2 && (strcmp("-h",argIn[1]) == 0 || strcmp ("--help",argIn[1]) == 0 )) {
usage(1);
};
time(&g_statsAll.timeStart);
///////////////////////////////////////////////////////////////////////
///////////////////////////////////////////// Parameters
Parameters P; //all parameters
P.inputParameters(argInN, argIn);
*(P.inOut->logStdOut) << timeMonthDayTime(g_statsAll.timeStart) << " ..... started STAR run\n" <<flush;
//runMode
if ( P.runMode == "alignReads" || P.runMode == "soloCellFiltering" ){
//continue
} else if ( P.runMode=="genomeGenerate" ) {
{//normal genome generation
Genome genomeMain(P, P.pGe);
genomeMain.genomeGenerate();
};
if (P.pGe.transform.type>0) {//generate original genome, in addition to the transfomed generated above
P.pGe.transform.type = 0;
P.pGe.transform.typeString = "None";
P.pGe.transform.vcfFile = "-";
P.pGe.gDir += "/OriginalGenome/";
Genome genomeOrig(P, P.pGe);
genomeOrig.genomeGenerate();
};
sysRemoveDir (P.outFileTmp);
P.inOut->logMain << "DONE: Genome generation, EXITING\n" << flush;
exit(0);
} else if ( P.runMode == "liftOver" ) {
for (uint ii=0; ii<P.pGe.gChainFiles.size();ii++) {
Chain chain(P,P.pGe.gChainFiles.at(ii));
chain.liftOverGTF(P.pGe.sjdbGTFfile,P.outFileNamePrefix+"GTFliftOver_"+to_string(ii+1)+".gtf");
P.inOut->logMain << "DONE: lift-over of GTF file, EXITING\n" << flush;
exit(0);
};
} else {
P.inOut->logMain << "EXITING because of INPUT ERROR: unknown value of input parameter runMode=" <<P.runMode<<endl<<flush;
exit(1);
};
//transcripome placeholder
Transcriptome *transcriptomeMain=NULL;
//this will execute --runMode soloCellFiltering and exit
Solo soloCellFilter(P, *transcriptomeMain);
////////////////////////////////////////////////////////////////////////
///////////////////////////////// Genome
Genome genomeMain (P, P.pGe);
genomeMain.genomeLoad();
genomeMain.Var=new Variation(P, genomeMain.chrStart, genomeMain.chrNameIndex);
SjdbClass sjdbLoci;
if (P.sjdbInsert.pass1) {
Genome genomeMain1=genomeMain;//not sure if I need to create the copy - genomeMain1 below should not be changed
sjdbInsertJunctions(P, genomeMain, genomeMain1, sjdbLoci);
};
/////////////////////////////////////////////////////////////////////////////////////////////////START
if (P.runThreadN>1) {
g_threadChunks.threadArray=new pthread_t[P.runThreadN];
pthread_mutex_init(&g_threadChunks.mutexInRead, NULL);
pthread_mutex_init(&g_threadChunks.mutexOutSAM, NULL);
pthread_mutex_init(&g_threadChunks.mutexOutBAM1, NULL);
pthread_mutex_init(&g_threadChunks.mutexOutUnmappedFastx, NULL);
pthread_mutex_init(&g_threadChunks.mutexOutFilterBySJout, NULL);
pthread_mutex_init(&g_threadChunks.mutexStats, NULL);
pthread_mutex_init(&g_threadChunks.mutexBAMsortBins, NULL);
pthread_mutex_init(&g_threadChunks.mutexError, NULL);
};
g_statsAll.progressReportHeader(P.inOut->logProgress);
/////////////////////////////////////////////////////////////////////////////
//////////////////////////////////// 2-pass 1st pass
twoPassRunPass1(P, genomeMain, transcriptomeMain, sjdbLoci);
if ( P.quant.yes ) {//load transcriptome
transcriptomeMain=new Transcriptome(P);
};
//initialize Stats
g_statsAll.resetN();
time(&g_statsAll.timeStartMap);
*P.inOut->logStdOut << timeMonthDayTime(g_statsAll.timeStartMap) << " ..... started mapping\n" <<flush;
g_statsAll.timeLastReport=g_statsAll.timeStartMap;
//SAM headers
samHeaders(P, *genomeMain.genomeOut.g, *transcriptomeMain);
//initialize chimeric parameters here - note that chimeric parameters require samHeader
P.pCh.initialize(&P);
// this does not seem to work at the moment
// P.inOut->logMain << "mlock value="<<mlockall(MCL_CURRENT|MCL_FUTURE) <<"\n"<<flush;
// prepare chunks and spawn mapping threads
ReadAlignChunk *RAchunk[P.runThreadN];
for (int ii=0;ii<P.runThreadN;ii++) {
RAchunk[ii]=new ReadAlignChunk(P, genomeMain, transcriptomeMain, ii);
};
if (P.runRestart.type!=1)
mapThreadsSpawn(P, RAchunk);
if (P.outFilterBySJoutStage==1) {//completed stage 1, go to stage 2
P.inOut->logMain << "Completed stage 1 mapping of outFilterBySJout mapping\n"<<flush;
outputSJ(RAchunk,P);//collapse novel junctions
P.readFilesIndex=-1;
P.outFilterBySJoutStage=2;
if (P.outBAMcoord) {
for (int it=0; it<P.runThreadN; it++) {//prepare the unmapped bin
RAchunk[it]->chunkOutBAMcoord->coordUnmappedPrepareBySJout();
};
};
mapThreadsSpawn(P, RAchunk);
};
//close some BAM files
if (P.inOut->outBAMfileUnsorted!=NULL) {
bgzf_flush(P.inOut->outBAMfileUnsorted);
bgzf_close(P.inOut->outBAMfileUnsorted);
};
if (P.inOut->outQuantBAMfile!=NULL) {
bgzf_flush(P.inOut->outQuantBAMfile);
bgzf_close(P.inOut->outQuantBAMfile);
};
if (P.outBAMcoord && P.limitBAMsortRAM==0) {//make it equal ot the genome size
P.limitBAMsortRAM=genomeMain.nGenome+genomeMain.SA.lengthByte+genomeMain.SAi.lengthByte;
};
time(&g_statsAll.timeFinishMap);
*P.inOut->logStdOut << timeMonthDayTime(g_statsAll.timeFinishMap) << " ..... finished mapping\n" <<flush;
//no need for genome anymore, free the memory
genomeMain.freeMemory();
//aggregate output junctions
//collapse splice junctions from different threads/chunks, and output them
if (P.runRestart.type!=1)
outputSJ(RAchunk,P);
//solo counts
Solo soloMain(RAchunk,P,*RAchunk[0]->chunkTr);
soloMain.processAndOutput();
if ( P.quant.geCount.yes ) {//output gene quantifications
for (int ichunk=1; ichunk<P.runThreadN; ichunk++) {//sum counts from all chunks into 0th chunk
RAchunk[0]->chunkTr->quants->addQuants(*(RAchunk[ichunk]->chunkTr->quants));
};
RAchunk[0]->chunkTr->quantsOutput();
};
if (P.runThreadN>1 && P.outSAMorder=="PairedKeepInputOrder") {//concatenate Aligned.* files
RAchunk[0]->chunkFilesCat(P.inOut->outSAM, P.outFileTmp + "/Aligned.out.sam.chunk", g_threadChunks.chunkOutN);
};
bamSortByCoordinate(P, RAchunk, *genomeMain.genomeOut.g, soloMain);
//wiggle output
if (P.outWigFlags.yes) {
*(P.inOut->logStdOut) << timeMonthDayTime() << " ..... started wiggle output\n" <<flush;
P.inOut->logMain << timeMonthDayTime() << " ..... started wiggle output\n" <<flush;
string wigOutFileNamePrefix=P.outFileNamePrefix + "Signal";
signalFromBAM(P.outBAMfileCoordName, wigOutFileNamePrefix, P);
};
g_statsAll.writeLines(P.inOut->outChimJunction, P.pCh.outJunctionFormat, "#", STAR_VERSION + string(" ") + P.commandLine);
g_statsAll.progressReport(P.inOut->logProgress);
P.inOut->logProgress << "ALL DONE!\n"<<flush;
P.inOut->logFinal.open((P.outFileNamePrefix + "Log.final.out").c_str());
g_statsAll.reportFinal(P.inOut->logFinal);
*P.inOut->logStdOut << timeMonthDayTime(g_statsAll.timeFinish) << " ..... finished successfully\n" <<flush;
P.inOut->logMain << "ALL DONE!\n" << flush;
if (P.outTmpKeep=="None") {
sysRemoveDir (P.outFileTmp);
};
P.closeReadsFiles();//this will kill the readFilesCommand processes if necessary
//genomeMain.~Genome(); //need explicit call because of the 'delete P.inOut' below, which will destroy P.inOut->logStdOut
if (genomeMain.sharedMemory != NULL) {//need explicit call because this destructor will write to files which are deleted by 'delete P.inOut' below
delete genomeMain.sharedMemory;
genomeMain.sharedMemory = NULL;
};
delete P.inOut; //to close files
return 0;
};
|