I am attempting to run DEXSeq on an RNASeq data set to determine if there is differential splicing due to treatment condition. However, during the estimate dispersions step I get the following error: simpleLoess(y, x, w, span, degree, parametric, drop.square, normalize, : NA/NaN/Inf in foreign function call (arg 1) What would cause such an error given the experimental design and provided design formulas?
My experimental set up has 23 patients, and each patient has two conditions - time 0 (t0) and time 1 (t1).The sample metadata looks like the following:
PatientID SampleID Condition p1 p1-t0 t0 p1 p1-t1 t1 p2 p2-t0 t0 p2 p2-t1 t1 … … ... p23 p23-t0 t0 p23 p23-t1 t1
Due to patient variability, I would like to block by PatientID. Following the DEXSeq vignette example, I set up my design formula and null model as the following:
> design.dex <- ~ sample + exon + PatientID:exon + Condition:exon > null.model <- ~ sample + exon + PatientID:exon
I am using the wrapper function to complete the entire analysis with the following command:
> output <- DEXSeq(dexSeqdataset,fullModel = design.dex, reducedModel = null.model, BPPARAM = MulticoreParam(workers=8),fitExpToVar="Condition",quiet=FALSE)
> sessionInfo() R version 3.1.0 (2014-04-10) Platform: x86_64-unknown-linux-gnu (64-bit) locale: [1] LC_CTYPE=en_US.UTF-8 LC_NUMERIC=C [3] LC_TIME=en_US.UTF-8 LC_COLLATE=en_US.UTF-8 [5] LC_MONETARY=en_US.UTF-8 LC_MESSAGES=en_US.UTF-8 [7] LC_PAPER=en_US.UTF-8 LC_NAME=C [9] LC_ADDRESS=C LC_TELEPHONE=C [11] LC_MEASUREMENT=en_US.UTF-8 LC_IDENTIFICATION=C attached base packages: [1] parallel stats graphics grDevices utils datasets methods [8] base other attached packages: [1] DEXSeq_1.10.6 BiocParallel_0.6.1 [3] DESeq2_1.4.5 RcppArmadillo_0.4.300.8.0 [5] Rcpp_0.11.2 GenomicRanges_1.16.3 [7] GenomeInfoDb_1.0.2 IRanges_1.22.9 [9] Biobase_2.24.0 BiocGenerics_0.10.0 [11] BiocInstaller_1.14.2 loaded via a namespace (and not attached): [1] annotate_1.42.0 AnnotationDbi_1.26.0 BatchJobs_1.2 [4] BBmisc_1.7 biomaRt_2.20.0 Biostrings_2.32.0 [7] bitops_1.0-6 brew_1.0-6 checkmate_1.0 [10] codetools_0.2-8 DBI_0.2-7 digest_0.6.4 [13] fail_1.2 foreach_1.4.2 genefilter_1.46.1 [16] geneplotter_1.42.0 grid_3.1.0 hwriter_1.3 [19] iterators_1.0.7 lattice_0.20-29 locfit_1.5-9.1 [22] plyr_1.8.1 RColorBrewer_1.0-5 RCurl_1.95-4.1 [25] Rsamtools_1.16.1 RSQLite_0.11.4 sendmailR_1.1-2 [28] splines_3.1.0 statmod_1.4.20 stats4_3.1.0 [31] stringr_0.6.2 survival_2.37-7 tools_3.1.0 [34] XML_3.98-1.1 xtable_1.7-3 XVector_0.4.0 [37] zlibbioc_1.10.0
Thank you for your help Alejandro.
I just want to check before I attempt to run this again. The development version of DEXSeq would be version 1.11.14?
Just to follow up, I am attempting to use the development version of DEXSeq (version 1.11.14). I used the commands provided on the DEXSeq developmental version bioconductor page (http://bioconductor.jp/packages/3.0/bioc/html/DEXSeq.html) :
However, this still installs version 1.10.8. Furthermore, I attempted to download and install from source, but when I click on the source link I get the following message: "Forbidden - You don't have permission to access /packages/3.0/bioc/src/contrib/DEXSeq_1.11.14.tar.gz on this server."
How would you suggest I upgrade to this development version?
There are instructions for how to use the devel version of Bioconductor; be sure to keep your 'release' and 'devel' libraries separate, and be aware that you're making a more than trivial change to your set-up so expect the worst (though the update may well go smoothly). The next release of Bioconductor is scheduled for October 14.
And the link you provide is to a Japanese mirror; try visiting http://bioconductor.org/packages/3.0/bioc/html/DEXSeq.html (Bioconductor uses a service from Amazon to distribute packages globally, so there is less need than in the past to use a mirror).
Martin,
Thank you for your help. Following your instructions, I installed and loaded the developmental version of DEXSeq:
However, when I make the DEXSeq count data set I get the following error:
> dsds.IFNstim <- DEXSeqDataSetFromHTSeq(sample.metadata.DEX$countfile,sampleData=sample.metadata.DEX,design=design.dex,flattenedfile="~/Projects/External/IFNL3-HCV/Ensembl66_standard_genes+IFNL4_chrDEXSeq.gtf")
Error in ncol(mcols(dds)) :
error in evaluating the argument 'x' in selecting a method for function 'ncol': Error in (function (classes, fdef, mtable) :
unable to find an inherited method for function ‘mcols’ for signature ‘"DESeqDataSet"’
'mcols' is a method from S4Vectors which is loaded. Do you think this is an error due to using the developmental versions or is this something more systematic with my set up?
'Release' versions of Bioconductor have even numbers in the second field, e.g., the '22' in IRanges 1.22.10, whereas 'devel' versions are odd-numbered, '11' in DEXSeq. So you have a mixture of release and devel packages.
biocLite()
should update to devel;biocValid()
should confirm that things are ok.Martin,
Thanks again for your help. That worked great!
cfreije