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"The following N features requested have not been scaled" warning after running SCTransform
I have the following Seurat v5 object:
An object of class Seurat
15130 features across 215733 samples within 1 assay
Active assay: RNA (15130 features, 0 variable features)
130 layers present: counts.1.1, counts.10.1, counts.11.2, counts.12.2, counts.13.2, counts.14.2, counts.15.2, counts.16.2, counts.17.2, ... , counts.9.2,
I then run:
sub.seurat <- SCTransform(sub.seurat,
assay = "RNA",
new.assay.name = "SCT",
variable.features.n = 1500,
vars.to.regress = c("pMitochondrial_RNA", "Cycle.Score"))
And when I want to run RunPCA():
Warning message:
In PrepDR(object = object, features = features, verbose = verbose) :
The following 577 features requested have not been scaled (running reduction without them): CCL4, CHI3L1, SERPINE1, CCL5, CCL3, FN1, CNTNAP2, SKAP1, CLDN5, [... truncated]
The object that comes out of SCTransform()
:
An object of class Seurat
30260 features across 215733 samples within 2 assays
Active assay: SCT (15130 features, 1500 variable features)
3 layers present: counts, data, scale.data
1 other assay present: RNA
I can run:
>table(sub.seurat[["SCT"]]@var.features %in% rownames(sub.seurat[["SCT"]]@scale.data))
FALSE TRUE
577 923
> nrow(sub.seurat@[email protected])
[1] 6711
Is this a bugor intented behaviour?
Why is the scale.data
object 6711 genes in size and not 1500 like the requested variable.feature.n
?
Why are the Layers joined instead of separate?
Why are the 1500 variable genes not all included in the scale.data
?