tilschaef / scRNA-seq

From fastq to preprocessed counttable (for in-house CELSeq2 method), with Kallisto | Bustools workflow.
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scRNA-seq preprocessing

From Kallisto | Bustools workflow counttable to preprocessed Seurat object.

FlowChart_scRNA-seqPreprocessing_sm

Installation

  1. Install Miniconda.

  2. Install Mamba

    conda install mamba -n base -c conda-forge
  3. Download the newest release of scRNA-seq or clone the repository.

    git clone https://github.com/Rebecza/scRNA-seq.git
  4. Create & activate environment.

    cd scRNA-seq/envs
    mamba env create -f kb_scrna_R_seurat3.yaml
    conda activate kb_scrna_R_seurat3
  5. Install RStudio (optional)

    mamba install -n kb_scrna_R_seurat3 r::rstudio

    Note: RStudio should be started from within the conda environment.

  6. Running

    • RStudio: Open kb_seurat_pp.rmd -> Fill parameters -> Knit
    • Command-line: Run rmarkdown::render('kb_seurat_pp.rmd,....) from R console.

Running the alignment needed for scRNA-seq (and RNA Velocity)

The indexing and alignment for RNA Velocity can easily be performed with the scRNA-seq workflow of seq2science.