package org.broadinstitute.sting.analyzecovariates; import org.broadinstitute.sting.gatk.walkers.recalibration.*; import org.broadinstitute.sting.utils.PackageUtils; import org.broadinstitute.sting.utils.xReadLines; import org.broadinstitute.sting.utils.cmdLine.CommandLineProgram; import org.broadinstitute.sting.utils.cmdLine.Argument; import java.util.ArrayList; import java.util.List; import java.util.Map; import java.util.regex.Pattern; import java.io.*; /** * Created by IntelliJ IDEA. * User: rpoplin * Date: Dec 1, 2009 * * Create collapsed versions of the recal csv file and call R scripts to plot residual error versus the various covariates. */ class AnalyzeCovariatesCLP extends CommandLineProgram { ///////////////////////////// // Command Line Arguments ///////////////////////////// @Argument(fullName = "recal_file", shortName = "recalFile", doc = "The input recal csv file to analyze", required = false) private String RECAL_FILE = "output.recal_data.csv"; @Argument(fullName = "output_dir", shortName = "outputDir", doc = "The directory in which to output all the plots and intermediate data files", required = false) private String OUTPUT_DIR = "analyzeCovariates/"; @Argument(fullName = "path_to_Rscript", shortName = "Rscript", doc = "The path to your implementation of Rscript. For Broad users this is probably /broad/tools/apps/R-2.6.0/bin/Rscript", required = false) private String PATH_TO_RSCRIPT = "/broad/tools/apps/R-2.6.0/bin/Rscript"; @Argument(fullName = "path_to_resources", shortName = "resources", doc = "Path to resources folder holding the Sting R scripts.", required = false) private String PATH_TO_RESOURCES = "R/"; @Argument(fullName = "ignoreQ", shortName = "ignoreQ", doc = "Ignore bases with reported quality less than this number.", required = false) private int IGNORE_QSCORES_LESS_THAN = 5; @Argument(fullName = "numRG", shortName = "numRG", doc = "Only process N read groups. Default value: -1 (process all read groups)", required = false) private int NUM_READ_GROUPS_TO_PROCESS = -1; // -1 means process all read groups ///////////////////////////// // Private Member Variables ///////////////////////////// private AnalysisDataManager dataManager; // Holds the data HashMap, mostly used by TableRecalibrationWalker to create collapsed data hashmaps private ArrayList requestedCovariates; // List of covariates to be used in this calculation private final Pattern COMMENT_PATTERN = Pattern.compile("^#.*"); private final Pattern OLD_RECALIBRATOR_HEADER = Pattern.compile("^rg,.*"); private final Pattern COVARIATE_PATTERN = Pattern.compile("^ReadGroup,QualityScore,.*"); protected int execute() { // create the output directory where all the data tables and plots will go try { Process p = Runtime.getRuntime().exec("mkdir " + OUTPUT_DIR); } catch (IOException e) { throw new RuntimeException("Couldn't create directory: " + OUTPUT_DIR); } if( !OUTPUT_DIR.endsWith("/") ) { OUTPUT_DIR = OUTPUT_DIR + "/"; } if( !PATH_TO_RESOURCES.endsWith("/") ) { PATH_TO_RESOURCES = PATH_TO_RESOURCES + "/"; } // initialize all the data from the csv file and allocate the list of covariates System.out.println("Reading in input csv file..."); initializeData(); System.out.println("...Done!"); // output data tables for Rscript to read in System.out.println("Writing out intermediate tables for R..."); writeDataTables(); System.out.println("...Done!"); // perform the analysis using Rscript and output the plots System.out.println("Calling analysis R scripts and writing out figures..."); callRScripts(); System.out.println("...Done!"); return 1; } private void initializeData() { // Get a list of all available covariates List> classes = PackageUtils.getClassesImplementingInterface(Covariate.class); int lineNumber = 0; boolean foundAllCovariates = false; // Read in the covariates that were used from the input file requestedCovariates = new ArrayList(); try { for ( String line : new xReadLines(new File( RECAL_FILE )) ) { lineNumber++; if( COMMENT_PATTERN.matcher(line).matches() || OLD_RECALIBRATOR_HEADER.matcher(line).matches()) { ; // Skip over the comment lines, (which start with '#') } else if( COVARIATE_PATTERN.matcher(line).matches() ) { // The line string is either specifying a covariate or is giving csv data if( foundAllCovariates ) { throw new RuntimeException( "Malformed input recalibration file. Found covariate names intermingled with data in file: " + RECAL_FILE ); } else { // Found the covariate list in input file, loop through all of them and instantiate them String[] vals = line.split(","); for( int iii = 0; iii < vals.length - 3; iii++ ) { // There are n-3 covariates. The last three items are nObservations, nMismatch, and Qempirical boolean foundClass = false; for( Class covClass : classes ) { if( (vals[iii] + "Covariate").equalsIgnoreCase( covClass.getSimpleName() ) ) { foundClass = true; try { Covariate covariate = (Covariate)covClass.newInstance(); requestedCovariates.add( covariate ); } catch ( InstantiationException e ) { throw new RuntimeException( String.format("Can not instantiate covariate class '%s': must be concrete class.", covClass.getSimpleName()) ); } catch ( IllegalAccessException e ) { throw new RuntimeException( String.format("Can not instantiate covariate class '%s': must have no-arg constructor.", covClass.getSimpleName()) ); } } } if( !foundClass ) { throw new RuntimeException( "Malformed input recalibration file. The requested covariate type (" + (vals[iii] + "Covariate") + ") isn't a valid covariate option." ); } } } } else { // Found a line of data if( !foundAllCovariates ) { foundAllCovariates = true; // At this point all the covariates should have been found and initialized if( requestedCovariates.size() < 2 ) { throw new RuntimeException( "Malformed input recalibration file. Covariate names can't be found in file: " + RECAL_FILE ); } // Initialize any covariate member variables using the shared argument collection for( Covariate cov : requestedCovariates ) { cov.initialize( new RecalibrationArgumentCollection() ); } // Initialize the data hashMaps dataManager = new AnalysisDataManager( requestedCovariates.size() ); } addCSVData(line); // Parse the line and add the data to the HashMap } } } catch ( FileNotFoundException e ) { throw new RuntimeException("Can not find input file: " + RECAL_FILE); } catch ( NumberFormatException e ) { throw new RuntimeException("Error parsing recalibration data at line " + lineNumber + ". Perhaps your table was generated by an older version of CovariateCounterWalker."); } } private void addCSVData(String line) { String[] vals = line.split(","); // Check if the data line is malformed, for example if the read group string contains a comma then it won't be parsed correctly if( vals.length != requestedCovariates.size() + 3 ) { // +3 because of nObservations, nMismatch, and Qempirical throw new RuntimeException("Malformed input recalibration file. Found data line with too many fields: " + line + " --Perhaps the read group string contains a comma and isn't being parsed correctly."); } Object[] key = new Object[requestedCovariates.size()]; Covariate cov; int iii; for( iii = 0; iii < requestedCovariates.size(); iii++ ) { cov = requestedCovariates.get( iii ); key[iii] = cov.getValue( vals[iii] ); } // Create a new datum using the number of observations, number of mismatches, and reported quality score RecalDatum datum = new RecalDatum( Long.parseLong( vals[iii] ), Long.parseLong( vals[iii + 1] ), Double.parseDouble( vals[1] ), 0.0 ); // Add that datum to all the collapsed tables which will be used in the sequential calculation dataManager.addToAllTables( key, datum, IGNORE_QSCORES_LESS_THAN ); } private void writeDataTables() { int numReadGroups = 0; // for each read group for( Object readGroupKey : dataManager.getCollapsedTable(0).data.keySet() ) { if(NUM_READ_GROUPS_TO_PROCESS == -1 || ++numReadGroups <= NUM_READ_GROUPS_TO_PROCESS) { String readGroup = readGroupKey.toString(); RecalDatum readGroupDatum = (RecalDatum) dataManager.getCollapsedTable(0).data.get(readGroupKey); System.out.print("Writing out data tables for read group: " + readGroup + "\twith " + readGroupDatum.getNumObservations() + " observations" ); System.out.println("\tand aggregate residual error = " + String.format("%.3f", readGroupDatum.empiricalQualDouble(0) - readGroupDatum.getEstimatedQReported())); // for each covariate for( int iii = 1; iii < requestedCovariates.size(); iii++ ) { Covariate cov = requestedCovariates.get(iii); // Create a PrintStream PrintStream output = null; try { output = new PrintStream(new FileOutputStream(OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat")); } catch (FileNotFoundException e) { System.err.println("Can't create file: " + OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat"); System.exit(-1); } // Output the header output.println("Covariate\tQreported\tQempirical\tnMismatches\tnBases"); for( Object covariateKey : ((Map)dataManager.getCollapsedTable(iii).data.get(readGroupKey)).keySet()) { output.print( covariateKey.toString() + "\t" ); // Covariate RecalDatum thisDatum = (RecalDatum)((Map)dataManager.getCollapsedTable(iii).data.get(readGroupKey)).get(covariateKey); output.print( String.format("%.3f", thisDatum.getEstimatedQReported()) + "\t" ); // Qreported output.print( String.format("%.3f", thisDatum.empiricalQualDouble(0)) + "\t" ); // Qempirical output.print( thisDatum.getNumMismatches() + "\t" ); // nMismatches output.println( thisDatum.getNumObservations() ); // nBases } // Close the PrintStream output.close(); } } else { break; } } } private void callRScripts() { int numReadGroups = 0; // for each read group for( Object readGroupKey : dataManager.getCollapsedTable(0).data.keySet() ) { if(NUM_READ_GROUPS_TO_PROCESS == -1 || ++numReadGroups <= NUM_READ_GROUPS_TO_PROCESS) { String readGroup = readGroupKey.toString(); System.out.println("Analyzing read group: " + readGroup); Process p = null; // for each covariate for( int iii = 1; iii < requestedCovariates.size(); iii++ ) { Covariate cov = requestedCovariates.get(iii); try { if( iii == 1 ) { // Analyze reported quality p = Runtime.getRuntime().exec(PATH_TO_RSCRIPT + " " + PATH_TO_RESOURCES + "plot_residualError_QualityScoreCovariate.R" + " " + OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat" + " " + IGNORE_QSCORES_LESS_THAN); // The third argument is the Q scores that should be turned pink in the plot because they were ignored } else { // Analyze all other covariates p = Runtime.getRuntime().exec(PATH_TO_RSCRIPT + " " + PATH_TO_RESOURCES + "plot_residualError_OtherCovariate.R" + " " + OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat" + " " + cov.getClass().getSimpleName().split("Covariate")[0]); // The third argument is the name of the covariate in order to make the plots look nice } } catch (IOException ex) { try { Thread.sleep(1600); // wait for 1.6 seconds and then try to spawn the process again if( iii == 1 ) { // Analyze reported quality p = Runtime.getRuntime().exec(PATH_TO_RSCRIPT + " " + PATH_TO_RESOURCES + "plot_residualError_QualityScoreCovariate.R" + " " + OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat" + " " + IGNORE_QSCORES_LESS_THAN); // The third argument is the Q scores that should be turned pink in the plot because they were ignored } else { // Analyze all other covariates p = Runtime.getRuntime().exec(PATH_TO_RSCRIPT + " " + PATH_TO_RESOURCES + "plot_residualError_OtherCovariate.R" + " " + OUTPUT_DIR + readGroup + "." + cov.getClass().getSimpleName()+ ".dat" + " " + cov.getClass().getSimpleName().split("Covariate")[0]); // The third argument is the name of the covariate in order to make the plots look nice } } catch (InterruptedException e) { e.printStackTrace(); System.exit(-1); } catch (IOException e) { e.printStackTrace(); System.exit(-1); } } } } else { break; } } } } public class AnalyzeCovariates { public static void main(String args[]) { AnalyzeCovariatesCLP clp = new AnalyzeCovariatesCLP(); CommandLineProgram.start( clp, args ); System.exit(0); } }