Methanol Fixation of Cells for Single Cell RNA Sequencing

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

CG000136_SamplePrepDemonstratedProtocol - MethanolFixation_RevB.pdf

This Demonstrated Protocol describes best practices and general protocols for methanol fixation of cells, storing, rehydrating and recovering fixed cells from single cell suspensions in preparation for use in 10x GenomicsĀ® Single Cell Protocols. Minimizing the presence of cellular 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 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 Support site support.10xgenomics.com.