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SoupX tutorial: meta.data nCount_RNA contains information from previous RNA assay #18

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KriBaLin opened this issue Aug 8, 2023 · 0 comments

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@KriBaLin
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KriBaLin commented Aug 8, 2023

Hi,
thank you a lot for your SoupX tutorial. I find the video and the code very helpful during my own analysis since I am new to scRNAseq analysis. One thing that I stumbled upon is that the nCount_RNA is not "updated" when I adjust the RNA assay of the seurat object with the SoupX-filtered counts. I'm referring to line 94 (sobj@assays$RNA@counts <- out) of your .Rmd file in this repository.
I noticed this when I tried to check whether counts actually changed by
summary(sobj@meta.data$nCount_RNA == sobj@meta.data$nCount_original.counts)
and nothing had changed.
When I tried to overwrite the RNA assay with
sobj[["RNA"]] <- CreateAssayObject(counts = out) nothing changed. When I do this giving a new name to the assay, e.g. with
sobj[["RNAsoupx"]] <- CreateAssayObject(counts = out) it works.

I'm not sure if this is the right place to post my observation/question, maybe you have an idea how the sobj@meta.data$nCount_RNA information can be updated by maintaining the name "RNA" for this assay. I'm not sure whether giving the assay a new name could cause any problems during downstream analysis.

Thank you a lot :)

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