Nuclear Isolation of Post-Mortem Brain Tissue for snRNAseq

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

Jonathan Brenton, PhD

Key Personnel: Team Hardy, Team Wood,

University College London
Carlo Sala Frigerio

External Collaborator
Mina Ryten

Co-PI (Core Leadership): Team Hardy, Team Wood,

University College London
Regina H. Reynolds

External Collaborator
Emil Gustavsson, PhD

Key Personnel: Team Wood, Team Hardy,

University College London
Kylie Montgomery

External Collaborator

Nuclear Isolation of Post-Mortem Brain Tissue for snRNAseq

Output Details

Meet the Authors

Jonathan Brenton, PhD

Carlo Sala Frigerio

Mina Ryten

Regina H. Reynolds

Emil Gustavsson, PhD

Kylie Montgomery

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