Nuclear Isolation of Post-Mortem Brain Tissue for snRNAseq

By Jonathan Brenton, PhD, Carlo Sala Frigerio, Mina Ryten, Regina H. Reynolds, Emil Gustavsson, PhD and Kylie Montgomery
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Output Details

Description

This protocol details how to isolate nuclei from frozen brain tissue for single nuclear RNA sequencing using 10x Genomics GEM isolation using the Chromium accessory and Single Cell 3ʹ Reagent Kits. It is adapted from a protocol designed by Carlo Sala Frigerio. The nuclei are homogenised by hand, run through a density gradient, resuspended, counted and diluted to an appropriate concentration to run in the Chromium. This protocol is designed to be continued with the Chromium Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry Dual Index) to isolate nuclei in GEMs.

Identifier (DOI)
10.17504/protocols.io.yxmvm25xng3p/v1
Tags
  • 10X Genomics
  • Brain
  • Human
  • Nuclei isolation
  • Postemortem
  • snRNA-seq (Single-nuclear RNA-seq)
Team
Team Hardy
Program
Collaborative Research Network
Theme
PD Functional Genomics

Meet the Authors

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