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R script for 10X SingleCell data analyzed by Seurat.

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DrLGB/SingleCell_Analysis_R

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Step1 basied on analysis of SingleCells

R script for 10X SingleCell data analyzed by Seurat (include: QC、PCA、TSNE、UMAP、Cluster、FindAllMarkers、etc.)

Seurat_Single_Sample is used for analysing the single data from 10X cellranger. Seurat_multi_Sample is used for integrating the results from Seurat_Single_Sample.

Usage:

  Rscript Seurat_Single_Sample.R 10X_matrix_dir output_dir  Sample_name
  
  Rscript Seurat_multi_Sample.R A.rds,B.rds,C.rds,D.rds  output_dir
  
  (All Parameters in the R scripts can be changed by user.)

Step2 Advanced analysis of SingleCells

R scripts for 10x SingleCelll data include CytoTRACE, slingshot,SCENIC,copycat inferCNV,GSVA, GSEA analysis and hierarchical clustering etc.

CytoTRACE_method is used to predict differentiation potential of cells

The method of slingshot is used to analysis pseudotim trajectory

SCNICIC_method is used to analysis to transcription factor activity in single cells

copycat_method is used for CNV analysis to discrimination between malignant and nonmaligant cells

inferCNV_method is another method to analysis CNV

GSVA_method is used for pathway activity

GSEA_method is used to analysis metabolism activity

hierarchical_cluster_method is used for hierarchical clustering

Usage:

      Rscripts CytoTRACE_method.R 10x.seurat.rds outdir prefix
      Rscripts slingshot_method.R 10x.seurat.rds outdir prefix
      Rscripts SCENIC_method.R 10x.seurat.rds hg38_cistarget_db outdir prefix
      Rscripts copycat_method.R 10x.seurat.rds outdir prefix
      Rscripts inferCNV_method.R 10x.seurat.rds grch38.bed outdir prefix
      Rscripts GSVA_method.R 10x.seurat.rds pathway.db outdir prefix
      Rscripts GSEA_method.R 10x.seurat.rds metabolism.pathway.db outdir prefix
      Rscripts hierarchical_cluster_method.R 10x.seurat.rds  outdir prefix
      

Note: Before using this two R script, you need to install R(version >= 3.5) and some libraries(DoubletFinder、Seurat).

R: https://www.r-project.org/
BoubletFinder: https://github.com/chris-mcginnis-ucsf/DoubletFinder
Seurat: https://github.com/satijalab/seurat

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R script for 10X SingleCell data analyzed by Seurat.

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