Isolation of Nuclei for Single Cell RNA Sequencing

Demonstrated Protocol, Last Modified on January 12, 2018, Permalink

CG000124_SamplePrepDemonstratedProtocol - Nuclei_RevD.pdf

These Demonstrated Protocols describe best practices and general protocols for cell lysis, washing, debris removal, counting, and concentrating nuclei from both single cell suspensions and neural tissue in preparation for use in 10x GenomicsĀ® Single Cell Protocols. Minimizing the presence of nuclear aggregates, dead cells, cellular debris, cytoplasmic nucleic acids, and potential inhibitors of reverse transcription is critical to obtaining high quality data.

The Protocols described here are expected to be compatible with many, but not all, cell or tissue types. Additional optimization may be required for the preparation of cell or tissue types that are particularly sensitive to suspension composition or handling techniques. Preparation of single cells or isolation of nuclei direct from solid tissues or cryopreserved samples may also require additional optimization during dissociation and/or cell handling not covered here. For additional information on preparation of specific sample types, consult 10x Genomics Demonstrated Protocols available on the 10x Genomics Support site support.10xgenomics.com.