单细胞数据标准化和降维后,利用scDblFinder找双胞,但是一直报错。
object <- subset(object, subset = nFeature_RNA > 200 & nFeature_RNA < 6000 & nCount_RNA < 20000 & percent.mt < 5) # 200 < nFeature_RNA(单个细胞总基因数) < 6000; # nCount_RNA(单个细胞总表达量数) < 20000; # percent.mt(单个细胞内线粒体基因比例) < 5
# 进行seurat流程的预处理,三步标准化。
object <- NormalizeData(object)
object <- FindVariableFeatures(object, selection.method = "vst", nfeatures = 2000)
object <- ScaleData(object)
object <- RunPCA(object)
object <- RunUMAP(object, dims = 1:20)
object <- FindNeighbors(object, reduction = "pca", dims = 1:20)
object <- FindClusters(object, resolution = 0.5)
sce <- as.SingleCellExperiment(object) #将seurat对象转换成SingleCellExperiment 对象
sce <- scDblFinder(sce, dbr=0.1) # 设置10%的双联体预期比例(dbr=0.1)
