sci-RNA-seq
Hello,
I have sci-RNA-seq data. I applied standart Seuat pipeline. However, I have ‘dirty’ clusters on UMAP. So some clusters are mixed. What can I do to solve it? Can I use doublet removers for sci-RNA-seq data?
library(Seurat)
# Load the control dataset
ctrl.data <- Read10X(data.dir = "ctrl//")
# Initialize the Seurat object
ctrl <- CreateSeuratObject(counts = ctrl.data, project = "control", min.cells = 3, min.features = 200)
# filtering
ctrl <- subset(ctrl, subset = nFeature_RNA > 100 & nFeature_RNA < 5000)
# Normalizing the data
ctrl <- NormalizeData(ctrl, normalization.method = "LogNormalize", scale.factor = 10000)
# Identification of highly variable features (feature selection)
ctrl <- FindVariableFeatures(ctrl, selection.method = "vst", nfeatures = 2000)
# Scaling the data
ctrl <- ScaleData(ctrl)
# Perform linear dimensional reduction
ctrl <- RunPCA(ctrl)
# Determine the ‘dimensionality’ of the dataset
ElbowPlot(ctrl)
# Cluster cells
ctrl <- FindNeighbors(ctrl, dims = 1:15)
ctrl <- FindClusters(ctrl, resolution = c(0.3, 0.5, 0.7))
ctrl <- SetIdent(ctrl, value = "RNA_snn_res.0.7")
# Run non-linear dimensional reduction
ctrl <- RunUMAP(ctrl, dims = 1:15)
DimPlot(ctrl, reduction = "umap", label = T)
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