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FindMarkers on scATAC-seq using normalized Tn5 cuts? #1119

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ccshao opened this issue May 10, 2022 · 3 comments
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FindMarkers on scATAC-seq using normalized Tn5 cuts? #1119

ccshao opened this issue May 10, 2022 · 3 comments
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@ccshao
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ccshao commented May 10, 2022

I have a quite naive but anyway would like to confirm that the FindMarkers in Signac using the TF-IDF normlized Tn5 Cuts counts from Peak/Cell matrix provided by 10x, but NOT based on the fragment file?

Actually I saw a related note here:
#373 (comment)

The reason why I asked is that I think ArchR is using the fragments as the basic mearements (converted to tilematrix) and then performed the TF-IDF normalization. Differentially accessible peaks are computated subsequently. In the vignette I didn't find how they use Tn5 cuts at all. To me Signac and ArchR are quite different from the very beginning.

These two approaches also give us quite different results on clustering an UMAP.

Thanks!

@ccshao ccshao added the documentation Documentation help label May 10, 2022
@timoast
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timoast commented May 15, 2022

I think the issue you've linked to gives the answer to this question

Whether cut sites or fragments are counted to create the matrix depends on the method used. Cellranger counts cut sites, Signac currently counts fragments.

@timoast timoast closed this as completed May 15, 2022
@ccshao
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ccshao commented May 15, 2022

Cellranger counts cut sites, Signac currently counts fragments.

Now I get more confused. Wouldn't the data used in Signac be the cut sites from Peak/Cell matrix?
I mean, GetAssayData(object = pbmc, slot = "counts") will give the cuts in peak region?

@timoast
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timoast commented May 15, 2022

I'm referring to the FeatureMatrix() function in Signac counting fragments rather than cut sites

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