/* * Copyright (c) 2010 The Broad Institute * * Permission is hereby granted, free of charge, to any person * obtaining a copy of this software and associated documentation * files (the "Software"), to deal in the Software without * restriction, including without limitation the rights to use, * copy, modify, merge, publish, distribute, sublicense, and/or sell * copies of the Software, and to permit persons to whom the * Software is furnished to do so, subject to the following * conditions: * * The above copyright notice and this permission notice shall be * included in all copies or substantial portions of the Software. * * THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, * EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES * OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND * NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT * HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, * WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING * FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR * THE USE OR OTHER DEALINGS IN THE SOFTWARE. */ package org.broadinstitute.sting.analyzecovariates; import org.broadinstitute.sting.commandline.Argument; import org.broadinstitute.sting.commandline.CommandLineProgram; import org.broadinstitute.sting.commandline.Input; import org.broadinstitute.sting.gatk.walkers.recalibration.Covariate; import org.broadinstitute.sting.gatk.walkers.recalibration.RecalDatum; import org.broadinstitute.sting.gatk.walkers.recalibration.RecalibrationArgumentCollection; import org.broadinstitute.sting.utils.classloader.PluginManager; import org.broadinstitute.sting.utils.exceptions.DynamicClassResolutionException; import org.broadinstitute.sting.utils.text.XReadLines; import java.io.*; import java.util.ArrayList; import java.util.Collection; import java.util.Map; import java.util.regex.Pattern; /** * 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. */ public class AnalyzeCovariates extends CommandLineProgram { ///////////////////////////// // Command Line Arguments ///////////////////////////// @Input(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 maybe /broad/tools/apps/R-2.6.0/bin/Rscript", required = false) private String PATH_TO_RSCRIPT = "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 = "public/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 @Argument(fullName="max_quality_score", shortName="maxQ", required = false, doc="The integer value at which to cap the quality scores, default is 50") private int MAX_QUALITY_SCORE = 50; @Argument(fullName="max_histogram_value", shortName="maxHist", required = false, doc="If supplied, this value will be the max value of the histogram plots") private int MAX_HISTOGRAM_VALUE = 0; @Argument(fullName="do_indel_quality", shortName="indels", required = false, doc="If supplied, this value will be the max value of the histogram plots") private boolean DO_INDEL_QUALITY = false; ///////////////////////////// // 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 static final String EOF_MARKER = "EOF"; 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) { System.out.println("Couldn't create directory: " + OUTPUT_DIR); System.out.println("User is responsible for making sure the output directory exists."); } 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 0; } private void initializeData() { // Get a list of all available covariates Collection> classes = new PluginManager(Covariate.class).getPlugins(); 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() || line.equals(EOF_MARKER) ) { ; // 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 (Exception e) { throw new DynamicClassResolutionException(covClass, e); } } } 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, MAX_QUALITY_SCORE) - 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, MAX_QUALITY_SCORE)) + "\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() ) { Process p; if(++numReadGroups <= NUM_READ_GROUPS_TO_PROCESS || NUM_READ_GROUPS_TO_PROCESS == -1) { String readGroup = readGroupKey.toString(); System.out.println("Analyzing read group: " + readGroup); // for each covariate for( int iii = 1; iii < requestedCovariates.size(); iii++ ) { Covariate cov = requestedCovariates.get(iii); try { if (DO_INDEL_QUALITY) { p = Runtime.getRuntime().exec(PATH_TO_RSCRIPT + " " + PATH_TO_RESOURCES + "plot_indelQuality.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 p.waitFor(); } else { 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 + " " + MAX_QUALITY_SCORE + " " + MAX_HISTOGRAM_VALUE); // The third argument is the Q scores that should be turned pink in the plot because they were ignored p.waitFor(); } 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 p.waitFor(); } } } catch (InterruptedException e) { e.printStackTrace(); System.exit(-1); } catch (IOException e) { System.out.println("Fatal Exception: Perhaps RScript jobs are being spawned too quickly? One work around is to process fewer read groups using the -numRG option."); e.printStackTrace(); System.exit(-1); } } } else { // at the maximum number of read groups so break out break; } } } public static void main(String args[]) { try { AnalyzeCovariates clp = new AnalyzeCovariates(); start(clp, args); System.exit(CommandLineProgram.result); } catch (Exception e) { exitSystemWithError(e); } } }