/***************************************************************************** genomeCoverageMain.cpp (c) 2009 - Aaron Quinlan Hall Laboratory Department of Biochemistry and Molecular Genetics University of Virginia aaronquinlan@gmail.com Licenced under the GNU General Public License 2.0 license. ******************************************************************************/ #include "genomeCoverageBed.h" #include "version.h" using namespace std; // define our program name #define PROGRAM_NAME "bedtools genomecov" // define our parameter checking macro #define PARAMETER_CHECK(param, paramLen, actualLen) (strncmp(argv[i], param, min(actualLen, paramLen))== 0) && (actualLen == paramLen) // function declarations void genomecoverage_help(void); int genomecoverage_main(int argc, char* argv[]) { // our configuration variables bool showHelp = false; // input files string bedFile; string genomeFile; int max = INT_MAX; float scale = 1.0; float fragmentSize = 146; //Nucleosome :) bool haveBed = false; bool bamInput = false; bool haveGenome = false; bool startSites = false; bool bedGraph = false; bool bedGraphAll = false; bool eachBase = false; bool eachBaseZeroBased = false; bool obeySplits = false; bool haveScale = false; bool filterByStrand = false; bool pair_chip = false; bool haveSize = false; bool dUTP = false; bool only_5p_end = false; bool only_3p_end = false; bool add_gb_track_line = false; string gb_track_opts; string requestedStrand = "X"; // check to see if we should print out some help if(argc <= 1) showHelp = true; for(int i = 1; i < argc; i++) { int parameterLength = (int)strlen(argv[i]); if((PARAMETER_CHECK("-h", 2, parameterLength)) || (PARAMETER_CHECK("--help", 5, parameterLength))) { showHelp = true; } } if(showHelp) genomecoverage_help(); // do some parsing (all of these parameters require 2 strings) for(int i = 1; i < argc; i++) { int parameterLength = (int)strlen(argv[i]); if(PARAMETER_CHECK("-i", 2, parameterLength)) { if ((i+1) < argc) { haveBed = true; bedFile = argv[i + 1]; i++; } } else if(PARAMETER_CHECK("-ibam", 5, parameterLength)) { if ((i+1) < argc) { haveBed = true; bamInput = true; bedFile = argv[i + 1]; i++; } } else if(PARAMETER_CHECK("-g", 2, parameterLength)) { if ((i+1) < argc) { haveGenome = true; genomeFile = argv[i + 1]; i++; } } else if(PARAMETER_CHECK("-d", 2, parameterLength)) { eachBase = true; } else if(PARAMETER_CHECK("-dz", 3, parameterLength)) { eachBase = true; eachBaseZeroBased = true; } else if(PARAMETER_CHECK("-bg", 3, parameterLength)) { bedGraph = true; } else if(PARAMETER_CHECK("-bga", 4, parameterLength)) { bedGraphAll = true; } else if(PARAMETER_CHECK("-max", 4, parameterLength)) { if ((i+1) < argc) { max = atoi(argv[i + 1]); i++; } } else if(PARAMETER_CHECK("-scale", 6, parameterLength)) { if ((i+1) < argc) { haveScale = true; scale = atof(argv[i + 1]); i++; } } else if(PARAMETER_CHECK("-split", 6, parameterLength)) { obeySplits = true; } else if(PARAMETER_CHECK("-strand", 7, parameterLength)) { if ((i+1) < argc) { filterByStrand = true; requestedStrand = argv[i+1][0]; if (!(requestedStrand == "-" || requestedStrand == "+")) { cerr << "*****ERROR: invalid -strand value (" << requestedStrand << "). Allowed options are + or -" << endl; showHelp = true; } i++; } else { cerr << "*****ERROR: -strand options requires a value: + or -" << endl; showHelp = true; } } else if(PARAMETER_CHECK("-3", 2, parameterLength)) { only_3p_end = true; } else if(PARAMETER_CHECK("-5", 2, parameterLength)) { only_5p_end = true; } else if(PARAMETER_CHECK("-pc", 3, parameterLength)) { pair_chip = true; } else if(PARAMETER_CHECK("-fs", 3, parameterLength)) { if ((i+1) < argc) { haveSize = true; fragmentSize = atoi(argv[i + 1]); i++; } } else if(PARAMETER_CHECK("-du", 3, parameterLength)) { dUTP = true; } else if(PARAMETER_CHECK("-trackline", 10, parameterLength)) { add_gb_track_line = true; } else if(PARAMETER_CHECK("-trackopts", 10, parameterLength)) { if ((i+1) < argc) { add_gb_track_line = true; gb_track_opts = argv[i+1]; i++; } else { cerr << "*****ERROR: -trackopts options requires a value (UCSC/GB track definition parameters)" << endl; showHelp = true; } } else { cerr << endl << "*****ERROR: Unrecognized parameter: " << argv[i] << " *****" << endl << endl; showHelp = true; } } // make sure we have both input files if (!haveBed && !haveGenome && !bamInput) { cerr << endl << "*****" << endl << "*****ERROR: Need both a BED (-i) and a genome (-g) file. " << endl << "*****" << endl; showHelp = true; } // make sure we have both input files if (bamInput && haveGenome) { cerr << endl << "*****" << endl << "*****WARNING: Genome (-g) files are ignored when BAM input is provided. " << endl << "*****" << endl; } if (bedGraph && eachBase) { cerr << endl << "*****" << endl << "*****ERROR: Use -d/-dz or -bg, not both" << endl << "*****" << endl; showHelp = true; } if (bedGraphAll && eachBase) { cerr << endl << "*****" << endl << "*****ERROR: Use -d/-dz or -bga, not both" << endl << "*****" << endl; showHelp = true; } if (only_3p_end && only_5p_end) { cerr << endl << "*****" << endl << "*****ERROR: Use -3 or -5, not both " << endl << "*****" << endl; showHelp = true; } if ( (only_3p_end||only_5p_end) && obeySplits) { cerr << endl << "*****" << endl << "*****ERROR: Use -split can't be used with -3 or -5." << endl << "*****" << endl; showHelp = true; } if (add_gb_track_line && !(bedGraph||bedGraphAll)) { cerr << endl << "*****" << endl << "*****ERROR: Using -trackline requires bedGraph output (use -bg or -bga)." << endl << "*****" << endl; showHelp = true; } if (haveScale && !(bedGraph||bedGraphAll||eachBase)) { cerr << endl << "*****" << endl << "*****ERROR: Using -scale requires bedGraph output (use -bg or -bga) or per base depth (-d)." << endl << "*****" << endl; showHelp = true; } if (!showHelp) { BedGenomeCoverage *bc = new BedGenomeCoverage(bedFile, genomeFile, eachBase, startSites, bedGraph, bedGraphAll, max, scale, bamInput, obeySplits, filterByStrand, requestedStrand, only_5p_end, only_3p_end, pair_chip, haveSize, fragmentSize, dUTP, eachBaseZeroBased, add_gb_track_line, gb_track_opts); delete bc; } else { genomecoverage_help(); } return 0; } void genomecoverage_help(void) { cerr << "\nTool: bedtools genomecov (aka genomeCoverageBed)" << endl; cerr << "Version: " << VERSION << "\n"; cerr << "Summary: Compute the coverage of a feature file among a genome." << endl << endl; cerr << "Usage: " << PROGRAM_NAME << " [OPTIONS] -i -g " << endl << endl; cerr << "Options: " << endl; cerr << "\t-ibam\t\t" << "The input file is in BAM format." << endl; cerr << "\t\t\tNote: BAM _must_ be sorted by position" << endl << endl; cerr << "\t-d\t\t" << "Report the depth at each genome position (with one-based coordinates)." << endl; cerr << "\t\t\tDefault behavior is to report a histogram." << endl << endl; cerr << "\t-dz\t\t" << "Report the depth at each genome position (with zero-based coordinates)." << endl; cerr << "\t\t\tReports only non-zero positions." << endl; cerr << "\t\t\tDefault behavior is to report a histogram." << endl << endl; cerr << "\t-bg\t\t" << "Report depth in BedGraph format. For details, see:" << endl; cerr << "\t\t\tgenome.ucsc.edu/goldenPath/help/bedgraph.html" << endl << endl; cerr << "\t-bga\t\t" << "Report depth in BedGraph format, as above (-bg)." << endl; cerr << "\t\t\tHowever with this option, regions with zero " << endl; cerr << "\t\t\tcoverage are also reported. This allows one to" << endl; cerr << "\t\t\tquickly extract all regions of a genome with 0 " << endl; cerr << "\t\t\tcoverage by applying: \"grep -w 0$\" to the output." << endl << endl; cerr << "\t-split\t\t" << "Treat \"split\" BAM or BED12 entries as distinct BED intervals." << endl; cerr << "\t\t\twhen computing coverage." << endl; cerr << "\t\t\tFor BAM files, this uses the CIGAR \"N\" and \"D\" operations " << endl; cerr << "\t\t\tto infer the blocks for computing coverage." << endl; cerr << "\t\t\tFor BED12 files, this uses the BlockCount, BlockStarts, and BlockEnds" << endl; cerr << "\t\t\tfields (i.e., columns 10,11,12)." << endl << endl; cerr << "\t-strand\t\t" << "Calculate coverage of intervals from a specific strand." << endl; cerr << "\t\t\tWith BED files, requires at least 6 columns (strand is column 6). " << endl; cerr << "\t\t\t- (STRING): can be + or -" << endl << endl; cerr << "\t-pc\t\t" << "Calculate coverage of pair-end fragments." << endl; cerr << "\t\t\tWorks for BAM files only" << endl; cerr << "\t-fs\t\t" << "Force to use provided fragment size instead of read length" << endl; cerr << "\t\t\tWorks for BAM files only" << endl; cerr << "\t-du\t\t" << "Change strand af the mate read (so both reads from the same strand) useful for strand specific" << endl; cerr << "\t\t\tWorks for BAM files only" << endl; cerr << "\t-5\t\t" << "Calculate coverage of 5\" positions (instead of entire interval)." << endl << endl; cerr << "\t-3\t\t" << "Calculate coverage of 3\" positions (instead of entire interval)." << endl << endl; cerr << "\t-max\t\t" << "Combine all positions with a depth >= max into" << endl; cerr << "\t\t\ta single bin in the histogram. Irrelevant" << endl; cerr << "\t\t\tfor -d and -bedGraph" << endl; cerr << "\t\t\t- (INTEGER)" << endl << endl; cerr << "\t-scale\t\t" << "Scale the coverage by a constant factor." << endl; cerr << "\t\t\tEach coverage value is multiplied by this factor before being reported." << endl; cerr << "\t\t\tUseful for normalizing coverage by, e.g., reads per million (RPM)." << endl; cerr << "\t\t\t- Default is 1.0; i.e., unscaled." << endl; cerr << "\t\t\t- (FLOAT)" << endl << endl; cerr << "\t-trackline\t" << "Adds a UCSC/Genome-Browser track line definition in the first line of the output." << endl; cerr <<"\t\t\t- See here for more details about track line definition:" << endl; cerr <<"\t\t\t http://genome.ucsc.edu/goldenPath/help/bedgraph.html" << endl; cerr <<"\t\t\t- NOTE: When adding a trackline definition, the output BedGraph can be easily" << endl; cerr <<"\t\t\t uploaded to the Genome Browser as a custom track," << endl; cerr <<"\t\t\t BUT CAN NOT be converted into a BigWig file (w/o removing the first line)." << endl << endl; cerr << "\t-trackopts\t"<<"Writes additional track line definition parameters in the first line." << endl; cerr <<"\t\t\t- Example:" << endl; cerr <<"\t\t\t -trackopts 'name=\"My Track\" visibility=2 color=255,30,30'" << endl; cerr <<"\t\t\t Note the use of single-quotes if you have spaces in your parameters." << endl; cerr <<"\t\t\t- (TEXT)" << endl << endl; cerr << "Notes: " << endl; cerr << "\t(1) The genome file should tab delimited and structured as follows:" << endl; cerr << "\t " << endl << endl; cerr << "\tFor example, Human (hg19):" << endl; cerr << "\tchr1\t249250621" << endl; cerr << "\tchr2\t243199373" << endl; cerr << "\t..." << endl; cerr << "\tchr18_gl000207_random\t4262" << endl << endl; cerr << "\t(2) The input BED (-i) file must be grouped by chromosome." << endl; cerr << "\t A simple \"sort -k 1,1 > .sorted\" will suffice."<< endl << endl; cerr << "\t(3) The input BAM (-ibam) file must be sorted by position." << endl; cerr << "\t A \"samtools sort \" should suffice."<< endl << endl; cerr << "Tips: " << endl; cerr << "\tOne can use the UCSC Genome Browser's MySQL database to extract" << endl; cerr << "\tchromosome sizes. For example, H. sapiens:" << endl << endl; cerr << "\tmysql --user=genome --host=genome-mysql.cse.ucsc.edu -A -e \\" << endl; cerr << "\t\"select chrom, size from hg19.chromInfo\" > hg19.genome" << endl << endl; // end the program here exit(1); }