Protocol for mechanical nuclei dissociation and isolation from frozen tissue and organoids for single-nucleus RNA sequencing.

Authors

Marion Guillon, Margaux Brin, Laetitia Laurent, Anthony Flamier

Published in

STAR protocols. Volume 7. Issue 4. Pages 104855. Sep 28, 2026. Epub Sep 28, 2026.

Abstract

Preserving RNA integrity in complex tissues such as the brain is challenging. Here, we present a protocol for isolating intact nuclei from frozen tissue and organoids for single-nucleus RNA sequencing (snRNA-seq). We describe steps for mechanical dissociation, using the GentleMACS Dissociator, serial filtration, and magnetic bead-based nuclei isolation. We then detail nuclei quality control and counting. This approach yields pure single-nucleus suspensions suitable for downstream fixation, cryopreservation, or library preparation across diverse tissue types. For complete details on the use and execution of this protocol, please refer to Guillon et al.¹.

PMID:
42804364
Bibliographic data and abstract were imported from PubMed on 29 Sep 2026.

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