/* bigWigAverageOverBed - Compute average score of big wig over each bed, which may have introns. */ #include "common.h" #include "linefile.h" #include "hash.h" #include "localmem.h" #include "options.h" #include "verbose.h" #include "basicBed.h" #include "bigWig.h" #include "bits.h" char *bedOut = NULL; int sampleAroundCenter = 0; void usage() /* Explain usage and exit. */ { errAbort( "bigWigAverageOverBed - Compute average score of big wig over each bed, which may have introns.\n" "usage:\n" " bigWigAverageOverBed in.bw in.bed out.tab\n" "The output columns are:\n" " name - name field from bed, which should be unique\n" " size - size of bed (sum of exon sizes\n" " covered - # bases within exons covered by bigWig\n" " sum - sum of values over all bases covered\n" " mean0 - average over bases with non-covered bases counting as zeroes\n" " mean - average over just covered bases\n" "Options:\n" " -bedOut=out.bed - Make output bed that is echo of input bed but with mean column appended\n" " -sampleAroundCenter=N - Take sample at region N bases wide centered around bed item, rather\n" " than the usual sample in the bed item.\n" ); } static struct optionSpec options[] = { {"bedOut", OPTION_STRING}, {"sampleAroundCenter", OPTION_INT}, {NULL, 0}, }; void checkUniqueNames(struct bed *bedList) /* Make sure all names in bedList are unique */ { struct hash *hash = hashNew(16); struct bed *bed; for (bed = bedList; bed != NULL; bed = bed->next) { char *name = bed->name; if (hashLookup(hash, name) != NULL) errAbort("%s duplicated in input bed", name); else hashAdd(hash, name, NULL); } hashFree(&hash); } void addBigWigIntervalInfo(struct bbiFile *bbi, struct lm *lm, char *chrom, int start, int end, int *pSumSize, int *pSumCoverage, double *pSumVal) /* Read in interval from bigBed and add it sums. */ { struct bbiInterval *iv, *ivList = bigWigIntervalQuery(bbi, chrom, start, end, lm); *pSumSize += (end - start); for (iv = ivList; iv != NULL; iv = iv->next) { int cov1 = rangeIntersection(iv->start, iv->end, start, end); if (cov1 > 0) { *pSumCoverage += cov1; *pSumVal += cov1 * iv->val; } } } int countBlocks(struct bed *bedList, int fieldCount) /* Return the number of blocks in list, or if non-blocked beds, just number of beds. */ { if (fieldCount < 12) return slCount(bedList); int blockCount = 0; struct bed *bed; for (bed = bedList; bed != NULL; bed = bed->next) blockCount += bed->blockCount; return blockCount; } void optionallyPrintBedPlus(FILE *f, struct bed *bed, int fieldCount, double extra) /* Print BED to tab separated file plus an extra double-format column. */ { if (f != NULL) { bedOutputN(bed, fieldCount, f, '\t', '\t'); fprintf(f, "%g\n", extra); } } void averageFetchingEachBlock(struct bbiFile *bbi, struct bed *bedList, int fieldCount, FILE *f, FILE *bedF) /* Do the averaging fetching each block from bedList from bigWig. Fastest for short bedList. */ { struct lm *lm = lmInit(0); struct bed *bed; for (bed = bedList; bed != NULL; bed = bed->next) { int coverage = 0; double sum = 0.0; int size = 0; if (sampleAroundCenter > 0) { int center = (bed->chromStart + bed->chromEnd)/2; int left = center - (sampleAroundCenter/2); addBigWigIntervalInfo(bbi, lm, bed->chrom, left, left+sampleAroundCenter, &size, &coverage, &sum); } else { if (fieldCount < 12) addBigWigIntervalInfo(bbi, lm, bed->chrom, bed->chromStart, bed->chromEnd, &size, &coverage, &sum); else { int i; for (i=0; iblockCount; ++i) { int start = bed->chromStart + bed->chromStarts[i]; int end = start + bed->blockSizes[i]; addBigWigIntervalInfo(bbi, lm, bed->chrom, start, end, &size, &coverage, &sum); } } } /* Print out result, fudging mean to 0 if no coverage at all. */ double mean = 0; if (coverage > 0) mean = sum/coverage; fprintf(f, "%s\t%d\t%d\t%g\t%g\t%g\n", bed->name, size, coverage, sum, sum/size, mean); optionallyPrintBedPlus(bedF, bed, fieldCount, mean); } } int bedCmpChrom(const void *va, const void *vb) /* Compare strings such as chromosome names that may have embedded numbers, * so that chr4 comes before chr14 */ { const struct bed *a = *((struct bed **)va); const struct bed *b = *((struct bed **)vb); return cmpStringsWithEmbeddedNumbers(a->chrom, b->chrom); } struct bed *nextChromInList(struct bed *bedList) /* Return first bed in list that starts with another chromosome, or NULL if none. */ { char *chrom = bedList->chrom; struct bed *bed; for (bed = bedList->next; bed != NULL; bed = bed->next) if (!sameString(bed->chrom, chrom)) break; return bed; } void addBufIntervalInfo(double *valBuf, Bits *covBuf, int start, int end, int *pSumSize, int *pSumCoverage, double *pSumVal) /* Look at interval in buffers and add result to sums. */ { int size1 = end - start; *pSumSize += size1; int cov1 = bitCountRange(covBuf, start, size1); *pSumCoverage += cov1; int i; double sum1 = 0; for (i=start; ichrom; nextChrom = nextChromInList(bedList); verbose(2, "Processing %s\n", chrom); if (bigWigValsOnChromFetchData(chromVals, chrom, bbi)) { double *valBuf = chromVals->valBuf; Bits *covBuf = chromVals->covBuf; /* Loop through beds doing sums and outputting. */ for (bed = bedList; bed != nextChrom; bed = bed->next) { int size = 0, coverage = 0; double sum = 0.0; if (sampleAroundCenter > 0) { int center = (bed->chromStart + bed->chromEnd)/2; int left = center - (sampleAroundCenter/2); addBufIntervalInfo(valBuf, covBuf, left, left+sampleAroundCenter, &size, &coverage, &sum); } else { if (fieldCount < 12) { addBufIntervalInfo(valBuf, covBuf, bed->chromStart, bed->chromEnd, &size, &coverage, &sum); } else { int i; for (i=0; iblockCount; ++i) { int start = bed->chromStart + bed->chromStarts[i]; int end = start + bed->blockSizes[i]; addBufIntervalInfo(valBuf, covBuf, start, end, &size, &coverage, &sum); } } } /* Print out result, fudging mean to 0 if no coverage at all. */ double mean = 0; if (coverage > 0) mean = sum/coverage; fprintf(f, "%s\t%d\t%d\t%g\t%g\t%g\n", bed->name, size, coverage, sum, sum/size, mean); optionallyPrintBedPlus(bedF, bed, fieldCount, mean); } verboseDot(); } else { /* If no bigWig data on this chromosome, just output as if coverage is 0 */ for (bed = bedList; bed != nextChrom; bed = bed->next) { fprintf(f, "%s\t%d\t0\t0\t0\t0\n", bed->name, bedTotalBlockSize(bed)); optionallyPrintBedPlus(bedF, bed, fieldCount, 0); } } } verbose(1, "\n"); } void bigWigAverageOverBed(char *inBw, char *inBed, char *outTab) /* bigWigAverageOverBed - Compute average score of big wig over each bed, which may have introns. */ { struct bed *bedList; int fieldCount; bedLoadAllReturnFieldCount(inBed, &bedList, &fieldCount); checkUniqueNames(bedList); struct bbiFile *bbi = bigWigFileOpen(inBw); FILE *f = mustOpen(outTab, "w"); FILE *bedF = NULL; if (bedOut != NULL) bedF = mustOpen(bedOut, "w"); /* Count up number of blocks in file. It takes about 1/100th of of second to * look up a single block in a bigWig. On the other hand to stream through * the whole file setting a array of doubles takes about 30 seconds, so we change * strategy at 3,000 blocks. * I (Jim) usually avoid having two paths through the code like this, and am tempted * to always go the ~30 second chromosome-at-a-time way. On the other hand the block-way * was developed first, and it was useful to have both ways to test against each other. * (This found a bug where the chromosome way wasn't handling beds in chromosomes not * covered by the bigWig for instance). Since this code is not likely to change too * much, keeping both implementations in seems reasonable. */ int blockCount = countBlocks(bedList, fieldCount); verbose(2, "Got %d blocks, if >= 3000 will use chromosome-at-a-time method\n", blockCount); if (blockCount < 3000) averageFetchingEachBlock(bbi, bedList, fieldCount, f, bedF); else averageFetchingEachChrom(bbi, &bedList, fieldCount, f, bedF); carefulClose(&bedF); carefulClose(&f); } int main(int argc, char *argv[]) /* Process command line. */ { optionInit(&argc, argv, options); if (argc != 4) usage(); bedOut = optionVal("bedOut", bedOut); sampleAroundCenter = optionInt("sampleAroundCenter", sampleAroundCenter); bigWigAverageOverBed(argv[1], argv[2], argv[3]); return 0; }