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Neuroscience

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JoVE Journal Neuroscience
Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing
 

Isolation of Adult Spinal Cord Nuclei for Massively Parallel Single-nucleus RNA Sequencing

Article DOI: 10.3791/58413-v • 06:38 min • October 12th, 2018
October 12th, 2018 •
1, 1, 2, 3,4, 3, 1
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

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Summary
Automatic Translation

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.

Tags

Single-nucleus RNA Sequencing Spinal Cord Nuclei Isolation Method Central Nervous System Frozen Tissue Cell Type-specific Changes Mouse Euthanasia Inner Organs Vertebral Column Spinal Cord Extraction PBS Solution
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