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Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing
1Spinal Circuits and Plasticity Unit,National Institute of Neurological Disorders and Stroke, 2Bioinformatics Section, Information Technology Program,National Institute of Neurological Disorders and Stroke, 3Laboratory of Cochlear Development,National Institute on Deafness and Other Communication Disorders, 4Single Cell Analysis Facility,Frederick National Laboratory
Chapters
- 00:04Title
- 00:43Preparation of the Spinal Cord
- 02:03Detergent-mechanical Cell Lysis
- 03:25Homogenization and Sucrose Density Gradient
- 05:11Results: Representative Results of Nuclei Isolated from the Adult Mouse Spinal Cord
- 05:51Conclusion
Here, we present a protocol to rapidly isolate high-quality nuclei from the fresh or frozen tissue for downstream massively parallel RNA sequencing. We include detergent-mechanical and hypotonic-mechanical tissue disruption and cell lysis options, both of which can be used for isolation of nuclei.
