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

Abstract

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).

Keywords

• Cell Biology
• Cell isolation
• Single Cell
• Flow Cytometry/Mass Cytometry
• RNAseq
• Neuroscience