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URL: https://dx.doi.org/DOI:10.17504/protocols.io.bhpej5je

Authors: Sanjay Subramanian, Stacey Huppert

Summary: This protocol is used to isolate human liver nuclei from frozen samples in an effort to facilitate human nuclei extraction for use in single nuclear RNA sequencing and single cell ATAC sequencing without the need for fresh samples. This approach enables the ability to acquire signatures from all cell types, even those from fibrotic liver samples. A brief fixation step is included in order to preserve nuclear membrane integrity and prevent nuclear leakage. This protocol has been tested and works in frozen human liver samples weighing approximately 1 gram stored in cryopreservation medium. Single nuclear RNA seqencing was run using the 10x Chromium system.This project was supported in part by NIH P30 DK078392 (Gene Analysis Core) of the Digestive Diseases Research Core Center in Cincinnati, R01DK107553, and the Cincinnati Pediatric Cell Atlas Center funded by Cincinnati Children’s Research Foundation to Stacey S. Huppert

Affiliations: [Division of Gastroenterology, Hepatology & Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, United States], [Division of Gastroenterology, Hepatology & Nutrition, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, United States; Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH, United States]

Version: 1

Publication Date: 2020

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