@Matt I've now included the cuff output in the post - I think this is what you were looking for.
Hello,
I am having an issue with gene tracking in cummeRbund. Here is the cuff output:
> cuff
CuffSet instance with:
4 samples
23285 genes
30073 isoforms
25872 TSS
24748 CDS
139440 promoters
155232 splicing
122340 relCDS
But when I try to run gene tracking, the following happens:
> myGeneId <- "ENSMUSG00000045440"
> myGene <- getGene(cuff,myGeneId)
> myGene
CuffGene instance for gene ENSMUSG00000045440
Short name: ENSMUSG00000045440
Slots:
annotation
features
fpkm
repFpkm
diff
count
isoforms CuffFeature instance of size 1
TSS CuffFeature instance of size 0
CDS CuffFeature instance of size 0
> genetrack <- makeGeneRegionTrack(myGene)
Error in `[.data.frame`(features(object), , featCols) :
undefined columns selected
Therefore, I'm unable to visualize transcript-level features of my gene of interest. I am using an Ensembl gtf and have set options(ucscChromosomeNames=FALSE), but no luck. I have also tried using the cummeRbund cufflinks data from the sample directory a<-readCufflinks(system.file("extdata", package="cummeRbund")) but still no luck. I have no issue analyzing 'myGene' or any other cummeRbund feature, so I assume I've accessed the data correctly. Any ideas on what the problem might be?
Thanks.
1 answer
You might take a look at this post on SEQanswers. You still did not post your entire R script, as we do not know what your cuff data frame looks like, and I suspect that your issue arises because your are not reading into R the proper GTF output from cufflinks.
Yes, for me this was the solution: Make sure to pass "merged.gtf" to cummeRbund.readCufflinks(), if that GTF-file was also used in the previous analysis with CuffDiff.
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If you post your entire R script as well as explain what you expected to happen, that would be helpful.
have you tried using the recommended genes.gtf file downloable from the Tuxedo package website?