Entering edit mode
fawazfebin
▴
60
@fawazfebin-14053
Last seen 4.3 years ago
Hi
I was doing a gene ontology analysis on a set of differentially expressed genes. The goana function returns an error saying ," No genes found in universe". Here is the code I ran for differential expression analysis ('countdata' represents the dataframe containing counts of each gene in each of the samples) :
> gns <- select(org.Hs.eg.db, row.names(countdata), c("ENTREZID","SYMBOL"),"ENSEMBL") 'select()' returned 1:many mapping between keys and columns > gns <- gns[!duplicated(gns[,1]),] > all.equal(gns$ENSEMBL, row.names(countdata)) [1] TRUE >condition <- c(rep("Control",3),rep("Resistant",2)) y <- DGEList(countdata, genes = gns,group=condition) > countsPerMillion <- cpm(y) > keep <- which(rowSums(countCheck) >= 2) > y <- y[keep,] > y <- calcNormFactors(y, method="TMM") > plotMDS(y) > design=model.matrix(~condition,y$samples) > y <- estimateDisp(y,design) > plotBCV(y) > fit <- glmFit(y,design) > lrt <- glmLRT(fit,coef=2) > edgeR_result = topTags(lrt, n=24000, adjust.method="BH", sort.by="PValue", p.value=0.05) > deGenes=decideTestsDGE(lrt, adjust.method="BH", p.value=0.05, lfc=1) > summary(deGenes) [,1] -1 353 0 13399 1 348 > deGenes=row.names(lrt)[as.logical(deGenes)] > plotSmear(lrt,de.tags=deGenes) > go <- goana(lrt) Error in goana.default(de = DEGenes, universe = universe, ...) : No genes found in universe. > traceback() 5: stop("No genes found in universe") 4: goana.default(de = DEGenes, universe = universe, ...) 3: goana(de = DEGenes, universe = universe, ...) 2: goana.DGELRT(lrt) 1: goana(lrt) > sessionInfo() R version 3.3.2 (2016-10-31) Platform: x86_64-w64-mingw32/x64 (64-bit) Running under: Windows 7 x64 (build 7600) locale: [1] LC_COLLATE=English_United States.1252 LC_CTYPE=English_United States.1252 [3] LC_MONETARY=English_United States.1252 LC_NUMERIC=C [5] LC_TIME=English_United States.1252 attached base packages: [1] grid stats4 parallel stats graphics grDevices utils datasets [9] methods base other attached packages: [1] org.Hs.eg.db_3.4.0 AnnotationDbi_1.36.2 Biobase_2.34.0 [4] BiocInstaller_1.24.0 edgeR_3.16.5 limma_3.30.13 [7] rtracklayer_1.34.2 GenomicRanges_1.26.4 GenomeInfoDb_1.10.3 [10] IRanges_2.8.2 S4Vectors_0.12.2 fastcluster_1.1.24 [13] reshape2_1.4.2 ggplot2_2.2.1 RSQLite_2.0 [16] BiocGenerics_0.20.0 loaded via a namespace (and not attached): [1] bitops_1.0-6 matrixStats_0.52.2 bit64_0.9-7 [4] RColorBrewer_1.1-2 httr_1.3.1 tools_3.3.2 [7] R6_2.2.2 DBI_0.7 lazyeval_0.2.0 [10] colorspace_1.3-2 bit_1.1-12 cummeRbund_2.16.0 [13] scales_0.5.0 stringr_1.2.0 digest_0.6.12 [16] Rsamtools_1.26.2 XVector_0.14.1 pkgconfig_2.0.1 [19] htmltools_0.3.6 BSgenome_1.42.0 rlang_0.1.2 [22] BiocParallel_1.8.2 VariantAnnotation_1.20.3 RCurl_1.95-4.8 [25] magrittr_1.5 GO.db_3.4.0 Matrix_1.2-11 [28] Rcpp_0.12.13 munsell_0.4.3 stringi_1.1.5 [31] SummarizedExperiment_1.4.0 zlibbioc_1.20.0 plyr_1.8.4 [34] blob_1.1.0 lattice_0.20-35 Biostrings_2.42.1 [37] splines_3.3.2 GenomicFeatures_1.26.4 locfit_1.5-9.1 [40] biomaRt_2.30.0 XML_3.98-1.9 latticeExtra_0.6-28 [43] data.table_1.10.4 gtable_0.2.0 tibble_1.3.4 [46] GenomicAlignments_1.10.1 memoise_1.1.0
Kindly support.
Yes, goana needs to know where to find the Entrez Gene IDs, so you have to tell it the column name. By default it tries to use the row names which, in this case, aren't the right IDs.