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package pacbio;
import java.io.File;
import dna.Data;
import fileIO.ReadWrite;
import fileIO.TextFile;
import shared.Parse;
import shared.Shared;
/**
* @author Brian Bushnell
* @date Oct 2, 2012
*
*/
public class MakePacBioScript {
/**
Be sure to replace:
@BUILDNUM with a number
@DIRTY_INPUT with the PacBio file
@CLEAN_INPUT_1 with the Illumina file
@ORGANISM with the name of the organism (or whatever)
@NUMSLOTS with the number of slots requested
@TARGET_SIZE with an estimate of the genome size, in bases. Examples: 160000000 or 160m or 0.16g are equivalent.
@RAM with e.g. Xmx31g
@MAXRAM with e.g. Xmx220g
@SCRIPT with the output file, e.g. run.sh
@MERGEREF with a list of reference files, e.g. chrom1.fa,chrom2.fa,chrom3.fa
@MERGEDIRTY with a list of dirty files, e.g. subreads1.fa,subreads2.fa,subreads3.fa
@MERGECLEAN with a list of clean files, e.g. illumina1.fq,illumina2.fq,illumina3.fq
@EXTRA with extra files for Illumina error correction. e.g. extra=a.fq,b.fq,c.fq
Optional:
@MAXREADS with the max number of clean reads to use in phase 1 (the slowest phase)
@REFERENCE with a reference file (optional)
@REFBUILD with a number
*/
public static void main(String[] args){
if(args==null || args.length<1){
System.out.println("\nThis program generates a script for error-correcting PacBio reads using Illumina reads.\nSample command line:\n");
// System.out.println("java -ea -Xmx64m"+(Shared.WINDOWS ? "" : " -cp "+Data.ROOT)+" jgi.MakePacBioScript " +
// "dirty=subreads.fa clean=illumina.fq ref=ecoliRef.fa name=ecoli " +
// "out=run.sh template="+(Shared.WINDOWS ? "" : "/house/homedirs/b/bushnell/template/")+"cleanPacbioTemplate.sh " +
// "targetsize=5.4m threads=24 ram=31 maxram=100 noderam=256 build=-1 refbuild=-1 maxreads=-1");
// System.out.println("java -ea -Xmx64m"+(Shared.WINDOWS ? "" : " -cp "+Data.ROOT)+" jgi.MakePacBioScript " +
// "dirty=subreads.fa clean=illumina.fq ref=ecoliRef.fa name=ecoli " +
// "out=run.sh template="+(Shared.WINDOWS ? "" : "/house/homedirs/b/bushnell/template/")+"cleanPacbioTemplate.sh " +
// "targetsize=5.4m threads=24 noderam=256");
// System.out.println("\n\nOr to be concise:");
System.out.println("java -ea -Xmx64m"+(Shared.WINDOWS ? "" : " -cp "+Data.ROOT())+" jgi.MakePacBioScript " +
"d=subreads.fa c=illumina.fq tpl=template.sh ts=5.4m t=24 nm=256");
System.out.println("\n\nInput files can optionally be comma-separated lists of files, and absolute pathing can be used.");
System.out.println("All input files may be raw, gzipped, or bzipped as long as they have the correct file extension.");
System.out.println();
System.out.println("\n***** Required Parameters *****\n");
System.out.println("d=, dirty= \tPath to dirty (PacBio) reads. May be comma-delimited for multiple files.");
System.out.println("c=, clean= \tPath to clean (Illumina) reads. May be comma-delimited for multiple files.");
System.out.println("t=, threads= \tNumber of threads. Should equal the number of qsub slots or cores on the target machine.");
System.out.println("nm=, nodemem= \tPhysical memory (RAM) of target machine, in gigabytes.");
System.out.println("ts=, targetsize= \tEstimated size of target genome, in bases (k, m, or g may be used). Optional ONLY if a reference is supplied.");
System.out.println("\n***** Optional Parameters *****\n");
System.out.println("tpl=, template= \tPath to template for this script. Default is /house/homedirs/b/bushnell/template/cleanPacbioTemplate_ecc.sh");
System.out.println("mode= \tCan be specified instead of 'template='. Values are 'pacbio', 'assembly', or 'ccs'");
System.out.println("sort= \tTrue or false. Determines whether clean reads are sorted (alphabetically) and duplicates are removed.");
System.out.println("r=, ref= \tPath to reference fasta. May be comma-delimited for multiple files.");
System.out.println("o=, out= \tName of output script. Default is 'run.sh'.");
System.out.println("name= \tName of organism. Default is 'organism'.");
System.out.println("h=, hours= \tTime limit (in hours) for autogenerated qsub command line.");
System.out.println("m=,mem= \tAmount of heap memory for Java to use. Default is 31g; must be at least 10x number input PacBio bases." +
"\n \tNote! Two steps, Illumina error correction and site stacking, will ignore this and use all physical memory.");
System.out.println("b=,build= \tPrefix for index build number. Default is 2, yielding successively improved builds 2, 200, 201, 202, ... 208");
System.out.println("rb=,refbuild= \tReference build number. Default is 1.");
System.out.println("cp=,classpath= \tClasspath to the program. If unspecified, will be autodetected as "+
(Shared.WINDOWS ? "/house/homedirs/b/bushnell/beta19/" : Data.ROOT()));
// r=ref.fa o=run.sh
System.exit(0);
}
String dirty=null;
String clean=null;
String name="organism";
String targetsize=null;
String ref=null;
String template=null;
String output="run.sh";
String extra="";
String classpath=(Shared.WINDOWS ? "/house/homedirs/b/bushnell/beta19/" : Data.ROOT());
String sort_in="";
String sorted="sorted_topo#.txt.gz";
String sorted_out="sorted_topo1.txt.gz";
String mergeref=null;
String mergedirty=null;
String mergeclean=null;
String qsub=null;
String cleanecc="@ORGANISM_ecc_1.txt.gz";
String cleanbadecc="@ORGANISM_ecc_1_BAD.txt.gz";
String cleanallecc="@ORGANISM_ecc_1_ALL.txt.gz";
String mode="pacbio";
int build=2;
int threads=24;
int ram=31;
int maxram=-1;
int refbuild=-1;
int noderam=-1;
long maxReads=-1;
int runtime=499;
boolean ecc=false;
boolean sort=true;
for(int i=0; i<args.length; i++){
final String arg=args[i];
final String[] split=arg.split("=");
if(split.length!=2){
System.out.println("Wrong number of arguments for variable "+split[0]);
System.exit(0);
}
String a=split[0].toLowerCase();
String b=split[1];
if(b.equalsIgnoreCase("null")){b=null;}
if(a.equals("threads") || a.startsWith("slots") || a.equals("t")){
threads=Integer.parseInt(b);
}else if(a.equals("mode")){
mode=b;
}else if(a.startsWith("reads") || a.startsWith("maxreads") || a.equals("rd")){
maxReads=Parse.parseKMG(b);
}else if(a.startsWith("build") || a.startsWith("genome") || a.equals("b")){
build=Integer.parseInt(b);
String s=Data.chromFname(1, build);
if((new File(s)).exists()){System.out.println("Warning! Genome build "+b+" already exists at "+s);}
}else if(a.startsWith("refbuild") || a.startsWith("refgenome") || a.equals("rb")){
refbuild=Integer.parseInt(b);
String s=Data.chromFname(1, refbuild);
if((new File(s)).exists()){System.out.println("Warning! Genome build "+b+" already exists at "+s);}
}else if(a.startsWith("ram") || a.startsWith("mem") || a.equals("m")){
ram=Integer.parseInt(b.toLowerCase().replaceAll("g", ""));
}else if(a.startsWith("maxram") || a.startsWith("maxmem") || a.equals("mm")){
maxram=Integer.parseInt(b.toLowerCase().replaceAll("g", ""));
}else if(a.startsWith("noderam") || a.startsWith("nodemem") || a.equals("nm")){
if(b!=null){noderam=Integer.parseInt(b.toLowerCase().replaceAll("g", ""));}
}else if(a.equals("runtime") || a.equals("hours") || a.equals("time") || a.equals("h")){
runtime=Integer.parseInt(b.toLowerCase().replaceAll("h", ""));
}else if(a.startsWith("dirty") || a.startsWith("pacbio") || a.equals("d")){
dirty=b;
if(dirty.contains(",")){
mergedirty=dirty;
dirty="concatenatedDirtyFiles.fa.gz";
if((new File(dirty)).exists()){System.out.println("Warning! file already exists: "+dirty);}
}else{
if(!(new File(b)).exists()){System.out.println("Warning! No such file: "+b);}
}
}else if(a.startsWith("clean") || a.startsWith("illumina") || a.equals("c")){
clean=b;
if(clean.contains(",")){
String ext="fq";
if(clean.contains(".fasta,") || clean.contains(".fa,") || clean.contains(".fasta.gz,") || clean.contains(".fa.gz,")){ext="fa";}
else if(clean.contains(".txt,") || clean.contains(".txt.gz,")){ext="txt";}
mergeclean=clean;
clean="concatenatedCleanFiles."+ext+".gz";
if((new File(clean)).exists()){System.out.println("Warning! file already exists: "+clean);}
}else{
if(!(new File(b)).exists()){System.out.println("Warning! No such file: "+b);}
}
}else if(a.startsWith("name") || a.startsWith("organism")){
name=b;
if(name==null){name="organism";}
}else if(a.startsWith("size") || a.startsWith("targetsize") || a.equals("ts")){
targetsize=b;
}else if(a.equals("path") || a.equals("classpath") || a.equals("cp")){
classpath=b;
}else if(a.startsWith("template") || a.equals("tpl")){
if(b!=null){
template=b;
if(!(new File(b)).exists()){System.out.println("Warning! No such file: "+b);}
}
}else if(a.startsWith("extra") || a.equals("ex")){
extra=("extra="+b);
if(!b.contains(",") && !(new File(b)).exists()){System.out.println("Warning! No such file: "+b);}
}else if(a.startsWith("ref") || a.equals("r")){
ref=b;
if(ref.contains(",")){
mergeref=ref;
ref="concatenatedReferenceFiles.fa.gz";
if((new File(ref)).exists()){System.out.println("Warning! file already exists: "+ref);}
}else{
if(!(new File(b)).exists()){System.out.println("Warning! No such file: "+b);}
}
}else if(a.startsWith("out") || a.equals("o")){
output=b;
if((new File(b)).exists()){System.out.println("Warning! Outfile already exists: "+b);}
}else if(a.startsWith("ecc")){
ecc=Parse.parseBoolean(b);
}else if(a.equals("sort")){
sort=Parse.parseBoolean(b);
}else{
throw new RuntimeException("Unknown parameter "+args[i]);
}
}
if(template==null){
if(mode==null){mode="pacbio";}
mode=mode.toLowerCase();
if(mode.equals("pacbio") || mode.equals("pacbio_illumina")){
template=(Shared.WINDOWS ? "C:/workspace/prune/cleanPacbioTemplate_ecc.sh" : "/house/homedirs/b/bushnell/template/cleanPacbioTemplate_ecc_maxram.sh");
}else if(mode.equals("assembly") || mode.equals("assembly_illumina")
|| mode.equals("reference") || mode.equals("reference_illumina")){
template=(Shared.WINDOWS ? "C:/workspace/prune/correctReference.sh" : "/house/homedirs/b/bushnell/template/correctReference_maxram.sh");
}else if(mode.equals("ccs") || mode.startsWith("ccs_")){
throw new RuntimeException("TODO: Mode "+mode+" is unfinished.");
}else if(mode.equals("pacbio_ccs") || mode.endsWith("_ccs")){
throw new RuntimeException("TODO: Mode "+mode+" is unfinished.");
}
}
// assert(false) : mode+", "+template;
if(ecc){
if(sort){
sort_in=cleanecc;
sorted_out=sorted.replaceFirst("#", "1");
}else{
sorted_out=sorted=clean;
}
}else{
if(sort){
sort_in=clean;
sorted_out=sorted.replaceFirst("#", "1");
cleanecc=sorted_out;
cleanbadecc=sorted_out;
cleanallecc=sorted_out;
}else{
sorted_out=sorted=clean;
cleanecc=clean;
cleanbadecc=clean;
cleanallecc=clean;
}
}
assert(threads>0);
if(noderam<1){
if(threads<9){noderam=144;}
else if(threads<25){noderam=252;}//Changed due to crash at 217 GB on 24-core nodes.
else if(threads<33){noderam=512;}
else if(threads<41){noderam=1024;}
else{noderam=2048;}
System.out.println("Set noderam at "+noderam+"g");
}
String slotram;
if(noderam%threads==0){slotram=(noderam/threads)+"G";}
else{slotram=((noderam*990)/threads)+"M";}
if(noderam>0){
if(maxram<1){
maxram=(int)(noderam*(noderam>256 ? 0.83 : 0.85f));
System.out.println("Set maxram at "+maxram+"g");
}
}
if(ram>maxram){
ram=maxram;
System.out.println("Set ram at "+maxram+"g");
}
if("auto".equalsIgnoreCase(targetsize) || (targetsize==null && ref!=null)){
if(ref==null){throw new RuntimeException("Ref file must be specified for auto targetsize.");}
File f=new File(ref);
if(!f.exists()){throw new RuntimeException("Ref file must exist for auto targetsize.");}
if(f.exists()){
targetsize=""+new File(ref).length();
if(ref.endsWith(".gz") || ref.endsWith(".gzip") || ref.endsWith(".zip") || ref.endsWith(".bz2")){
TextFile tf=new TextFile(ref, false);
long x=1;
for(String s=tf.nextLine(); s!=null; s=tf.nextLine()){x+=s.length();}
tf.close();
targetsize=""+x;
}
}
}
if(ref!=null && refbuild<1){
if(build==1){refbuild=2;}
else{refbuild=1;}
}
if(dirty==null){throw new RuntimeException("No dirty file specified.");}
if(clean==null){throw new RuntimeException("No clean file specified.");}
if(targetsize==null){throw new RuntimeException("No targetsize specified.");}
if(template==null){throw new RuntimeException("No template file specified.");}
if(!new File(template).exists()){throw new RuntimeException("Template file "+template+" does not exist; please specify a different template.");}
if(build==refbuild){throw new RuntimeException("Build id and ref build id must differ.");}
if(build<1){throw new RuntimeException("No build id.");}
if(ref!=null && refbuild<1){throw new RuntimeException("No ref build id.");}
if(ref==null && refbuild>0 && !(new File(Data.chromFname(1, refbuild))).exists()){throw new RuntimeException("Ref build id specified, but no reference file.");}
String[] lines;
{
TextFile tf=new TextFile(template, false);
lines=tf.toStringLines();
}
StringBuilder sb=new StringBuilder();
for(int i=0; i<lines.length; i++){
String s=lines[i];
boolean eccline=s.contains("?ecc?");
boolean sortline=s.contains("?sort?");
boolean refline=s.contains("?ref?");
boolean mergeline=s.contains("?ref?");
boolean optional=(!eccline && !sortline && !refline && !mergeline && s.startsWith("#?")); //Optional for some other reason
if(eccline){
s=s.replaceAll("\\?ecc\\?", "");
while(ecc && s.startsWith("#")){s=s.substring(1);}
}
if(sortline){
s=s.replaceAll("\\?sort\\?", "");
while(sort && s.startsWith("#")){s=s.substring(1);}
}
if(refline){
s=s.replaceAll("\\?ref\\?", "");
while(refbuild>0 && s.startsWith("#")){s=s.substring(1);}
}
if(!s.startsWith("#")){
if((eccline && !ecc) || (sortline && !sort) || (refline && refbuild<1)){s="#"+s;}
}
if(optional){
optional=true;
s=s.substring(2);
}
if((s.contains("@MAXRAM") && maxram>31) || (s.contains("@RAM") && ram>31)){
s=s.replace("-XX:+UseCompressedOops ", "");
}
s=s.replace("@CLEAN_ECC_1", cleanecc);
s=s.replace("@CLEAN_BAD_ECC_1", cleanbadecc);
s=s.replace("@CLEAN_ALL_ECC_1", cleanallecc);
s=s.replace("@SORT_IN", sort_in);
s=s.replace("@SORTED_OUT", sorted_out);
s=s.replace("@SORTED", sorted);
s=s.replace("@SLOTRAM", slotram);
s=s.replace("@BUILDNUM", ""+build);
s=s.replace("@DIRTY_INPUT", dirty);
s=s.replace("@CLEAN_INPUT_1", clean);
s=s.replace("@ORGANISM", name);
s=s.replace("@NUMSLOTS", ""+threads);
s=s.replace("@TARGET_SIZE", targetsize);
s=s.replace("@RAM", "-Xmx"+ram+"g");
s=s.replace("@MAXRAM", "-Xmx"+maxram+"g");
s=s.replace("@MAXREADS", ""+maxReads);
s=s.replace("@SCRIPT", (output==null ? "run.sh" : output));
s=s.replace("@EXTRA", extra);
s=s.replace("@RUNTIME", ""+runtime);
s=s.replace("@CLASSPATH", classpath);
if(s.contains("@REFBUILD")){
if(refbuild<1){
s="#"+s;
}else{
s=s.replace("@REFBUILD", ""+refbuild);
}
}
if(s.contains("@REFERENCE")){
if(ref==null){
s="#"+s;
}else{
s=s.replace("@REFERENCE", ref);
}
}
if(s.contains("@MERGECLEAN")){
if(mergeclean==null){
s="#"+s;
}else{
s=s.replace("@MERGECLEAN", mergeclean);
}
}
if(s.contains("@MERGEDIRTY")){
if(mergedirty==null){
s="#"+s;
}else{
s=s.replace("@MERGEDIRTY", mergedirty);
}
}
if(s.contains("@MERGEREF")){
if(mergeref==null){
s="#"+s;
}else{
s=s.replace("@MERGEREF", mergeref);
}
}
while(s.startsWith("##")){s=s.substring(1);}
assert(s==null || s.length()<1 || s.startsWith("#") || !s.contains("@")) : s;
if(s!=null && !s.startsWith("#//")){sb.append(s).append('\n');}
if(qsub==null && s.contains("export task") && s.contains("qsub")){
qsub=s;
}
}
if(output==null){
System.out.println(sb);
}else{
ReadWrite.writeString(sb, output, false);
System.out.println("Wrote "+output);
if(qsub!=null){
while(qsub.startsWith("#")){qsub=qsub.substring(1);}
System.out.println("The script can be executed on Genepool with the following command:\n\n"+qsub.trim());
}
}
}
}
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