Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays

Ruth M Williams; Tatjana Sauka-Spengler

doi:10.1016/j.xpro.2021.100414

Dissociation of chick embryonic tissue for FACS and preparation of isolated cells for genome-wide downstream assays

STAR Protoc. 2021 Mar 27;2(2):100414. doi: 10.1016/j.xpro.2021.100414. eCollection 2021 Jun 18.

Authors

Ruth M Williams¹, Tatjana Sauka-Spengler¹

Affiliation

¹ University of Oxford, MRC Weatherall Institute of Molecular Medicine, Radcliffe Department of Medicine, Oxford OX3 9DS, UK.

Abstract

In order to process samples by fluorescence-activated cell sorting (FACS), it is essential to obtain a single-cell suspension of dissociated cells. Numerous protocols and commercial reagents are available; however, each requires optimization for specific tissue types. Here, we describe an optimized protocol for dissociating dissected chick embryos across a broad span of developmental stages. We also provide protocols for processing targeted cell populations isolated using FACS for ATAC-seq, RNA-seq, and chromatin immunoprecipitation. For complete details on the use and execution of this protocol, please refer to Ling and Sauka-Spengler (2019) and Williams et al. (2019).

Keywords: Genomics; Model Organisms; Molecular Biology; Sequencing; Single Cell.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques / methods*
Cells, Cultured
Chick Embryo / cytology*
Chromatin Immunoprecipitation Sequencing / methods*
Flow Cytometry / methods*
Genomics
RNA-Seq

Grants and funding

G0902418/MRC_/Medical Research Council/United Kingdom