Dissociation of intact adult mouse cortical projection neurons for single-cell RNA-seq

Summary: This protocol provides an improved pipeline for dissociating intact projection neurons from adult mouse cortex for applications including droplet and plate-based single-cell RNA sequencing, qPCR, immunocytochemistry, and long-term in vitro cell culture. This protocol provides a robust and r...

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Bibliographic Details
Main Authors: Noa Golan, William B. Cafferty
Format: Article
Language:English
Published: Elsevier 2021-12-01
Series:STAR Protocols
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S266616672100647X
Description
Summary:Summary: This protocol provides an improved pipeline for dissociating intact projection neurons from adult mouse cortex for applications including droplet and plate-based single-cell RNA sequencing, qPCR, immunocytochemistry, and long-term in vitro cell culture. This protocol provides a robust and reproducible dissociation pipeline that uses exclusively off-the-shelf reagents, not requiring the use of expensive dissociation kits. The unique incubation steps, in combination with the FACS gating strategy, results in unparalleled enrichment for intact cortical neurons from the adult brain.For complete details on the use and execution of this protocol, please refer to Golan et al. (2021).
ISSN:2666-1667