Question: Can I isolate nuclei from frozen tissue for gene expression profiling?
Answer: Unfortunately, we do not have direct experience isolating nuclei from frozen tissue for Single Cell gene expression profiling.
We do have a Demonstrated Protocol for the isolation of nuclei from fresh neural tissue: https://support.10xgenomics.com/single-cell-gene-expression/index/doc/demonstrated-protocol-isolation-of-nuclei-for-single-cell-rna-sequencing
This protocol describes the best practices and general protocols for cell lysis, washing, debris removal, counting, and concentrating nuclei in preparation for use in the 10x Single Cell protocol.
Isolation of nuclei from frozen tissue may require additional optimization during dissociation and/or cell handling not covered in this protocol.
A customer-developed protocol is available on the 10x Community site, "'Frankenstein’ protocol for nuclei isolation from fresh and frozen tissue."
Note: Customer-Developed protocols are provided for general information only and are NOT directly supported, endorsed, or certified by 10x Genomics.
Products: Single Cell 3', VDJ