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This repository contains code to analyze single-nuclei RNA sequencing data from the ventrobasal thalamus and principal sensory trigeminal nucleus in neonatal mice. It performs quality control, clustering, and identification of marker genes to characterize cell types and their interconnectivity during early whisker pathway development.

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Harkany-Lab/Hevesi_2023

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Hevesi_2023

  1. System requirements
  • All software dependencies and operating systems (including version numbers) are available in the methods section
  • Archived version of this repo has been tested on x86_64-pc-linux-gnu (64-bit) platform running under Ubuntu 22.04.1 LTS with R version 4.2.2 and Python version 3.8.8
  • GPU is required to run ambient RNA removal with CellBender
  1. Installation guide
  • Installation is available via Snakemake framework infrastructure. The Snakefile pipeline is in the project root. Before run: 1) You need to edit path to the cellranger environment file on your computer ($CELLRANGER). 2) Be sure that Apptainer (former Singularity is installed and available in $PATH).
  • Typical install time on a "normal" desktop computer should be within a hour that you need to install cellranger, Anaconda, Apptainer/Singularity, to create snakemake conda environment and to download all configured container-images.
  1. Demo
  • We include preprocessed data that one can use to reproduce panels of the figure 2 related to snRNA-seq data from the manuscript.
  • It is expected that output images can slightly differ due to random factor native to dimension reduction technics.
  • Expected run time for demo on a "normal" desktop computer is within a hour.
  1. Instructions for use
  • To run the demo on preprocessed data you need to source code/render.R file, which will rerun rendering of Rmarkdown analysis notebooks for figure 2 via workflowr package.
  • To reproduce whole pipeline you would need to modify and run command snakemake -f --use-singularity -prk --cores <FILL-IN-N-CORES> --resources nvidia_gpu=1 --singularity-args "--nv --bind <FILL-IN-PATH-TO-CLONED-REPO>" --rerun-incomplete --retries 1 --resources mem_mb=<FILL-IN-MEM>

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This repository contains code to analyze single-nuclei RNA sequencing data from the ventrobasal thalamus and principal sensory trigeminal nucleus in neonatal mice. It performs quality control, clustering, and identification of marker genes to characterize cell types and their interconnectivity during early whisker pathway development.

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