Question: countsFromAbundance for gene-level RSEM import in tximport
0
gravatar for celi0001
16 months ago by
celi00010
celi00010 wrote:

Hi all,

I’ve used tximport to import RSEM's “*.genes.results”  files and I’ve found that tximport uses the values from RSEM’s "expected counts" regardless of the value specified to the argument “countsFromAbundance” (i.e. “lengthScaledTPM”, “scaledTPM” or “no”). Would I have to correct the expected count values for the average transcript length per gene before I perform a differential gene expression analysis with limma-voom

Thanks, 

Celine

tximport rsem • 290 views
ADD COMMENTlink modified 16 months ago by Michael Love25k • written 16 months ago by celi00010
Answer: countsFromAbundance for gene-level RSEM import in tximport
0
gravatar for Michael Love
16 months ago by
Michael Love25k
United States
Michael Love25k wrote:

Thanks for noting this. If you use the latest version of tximport you can provide the isoform files to tximport(), and then you can specify countsFromAbundance of "lengthScaledTPM" or "scaledTPM". I guess I could build out more support to do this from the gene-level files specifically for RSEM, but I like having the clean interface that is mostly built around importing transcript-level data (and we now support RSEM).

I've added a warning to tximport() now if countsFromAbundance != "no" and txIn=FALSE, because it is just importing the data from the gene-level files, and not replacing the counts that RSEM computed.

ADD COMMENTlink written 16 months ago by Michael Love25k

I see! Thanks for the quick reply!

ADD REPLYlink written 16 months ago by celi00010

Just realised that, anyway, I should have in fact been using the counts without gene length normalisation for the DEG analysis.

ADD REPLYlink modified 16 months ago • written 16 months ago by celi00010

Hi Love,

I've used tximport to import Salmon "quant.sf" file, and I found there are all "no" at line "countsFromAbundance" in the result file. But I don't know the meaning about "no" at line "countsFromAbundance". Could you please tell me some details? Furthermore, I also found the line "abundance" and "counts" in the result file. What's the exact meaning of "abundance"?

Thanks,

Pan

ADD REPLYlink written 15 months ago by melodypluto950

For the meaning of "no", you can check the help page, and see the argument countsFromAbundance

?tximport

Abundance is the relative abundance estimated by the tool. This is almost always in units of TPM. For more details on scaledTPM and lengthScaledTPM, take a look at the reference publication listed in ?tximport.

ADD REPLYlink modified 15 months ago • written 15 months ago by Michael Love25k
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