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Purification of mouse hepatic non-parenchymal cells or nuclei for use in ChIP-seq and other next-generation sequencing approaches.

Citation
Troutman, T. D., et al. “Purification Of Mouse Hepatic Non-Parenchymal Cells Or Nuclei For Use In Chip-Seq And Other Next-Generation Sequencing Approaches.”. Star Protocols, p. 100363.
Center UCSD-UCLA
Author Ty D Troutman, Hunter Bennett, Mashito Sakai, Jason S Seidman, Sven Heinz, Christopher K Glass
Keywords Cell isolation, ChIP-seq, chromatin immunoprecipitation (ChiP), Flow cytometry/mass cytometry, high-throughput screening, immunology, sequencing
Abstract

Significant advancements in understanding disease mechanisms can occur through combined analysis of next-generation sequencing datasets generated using purified cell populations. Here, we detail our optimized protocol for purification of mouse hepatic macrophages (or other liver non-parenchymal populations) suitable for use in various next-generation sequencing protocols. An alternative framework is described for sorting pre-fixed hepatic nuclei populations. This strategy has the advantage of rapidly preserving the nuclei and can facilitate success with ChIP-seq for more challenging molecules. For complete details on the use and execution of these protocols, please refer to Muse et al. (2018), Sakai et al. (2019), and Seidman et al. (2020).

Year of Publication
2021
Journal
STAR protocols
Volume
2
Issue
1
Number of Pages
100363
Date Published
03/2021
ISSN Number
2666-1667
DOI
10.1016/j.xpro.2021.100363
Alternate Journal
STAR Protoc
PMID
33748781
PMCID
PMC7960533
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