Entering edit mode
4 months ago
# After creating a merged seurat object using this code:
OVLT_merged <- merge(M_OVLT_Saline, y = c(M_OVLT_LPS, M_OVLT_PolyIC, F_OVLT_Saline, F_OVLT_LPS, F_OVLT_PolyIC), add.cell.ids = c("M_OVLT_Saline", "M_OVLT_LPS", "M_OVLT_PolyIC", "F_OVLT_Saline", "F_OVLT_LPS", "F_OVLT_PolyIC"), project = "OVLT")
# I then added the percent mitochondrial and RBC genes to the object,
OVLT_merged <- PercentageFeatureSet(OVLT_merged , pattern = "^mt-", col.name = "MT")
OVLT_merged <- PercentageFeatureSet(OVLT_merged , pattern = "^Hba", col.name = "RBC")
OVLT_merged <- PercentageFeatureSet(OVLT_merged , pattern = "^Hbb", col.name = "RBC")
# So I then could filter the data using:
OVLT_filtered <- subset(OVLT_merged, subset = RBC < 0.01 & MT < 5 & nFeature_RNA < 6000 & nFeature_RNA > 300)
# Which I then ran SCTransform v2 to normalize the data:
OVLT <- SCTransform(OVLT_filtered, vars.to.regress=c("MT","RBC"), vst.flavor = "v2", return.only.var.genes = FALSE, verbose = FALSE)
In return, I get 50 warning messages the first of which is specified below:
1: useNames = NA is deprecated. Instead, specify either useNames = TRUE or useNames = FALSE. Error in cat("1: useNames = NA is deprecated. Instead, specify either useNames = TRUE or useNames = FALSE.", : argument 2 (type 'S4') cannot be handled by 'cat'
Does anyone know what maybe going wrong?
Thank you so much!
Michelle
Please use the formatting bar (especially the
code
option) to present your post better. You can use backticks for inline code (`text` becomestext
), or use one of (a) the option highlighted in the image below/ (b) fenced code blocks for multi-line code. Fenced code blocks are useful in syntax highlighting. If your code has long lines with a single command, break those lines into multiple lines with proper escape sequences so they're easier to read and still run when copy-pasted. I've done it for you this time.Nothing to worry about. Seurat v5 release has introduced several versioning issues.