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

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).

SUBMITTER: Williams RM

PROVIDER: S-EPMC8039852 | biostudies-literature | 2021 Jun

REPOSITORIES: biostudies-literature

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

Williams Ruth M RM Sauka-Spengler Tatjana T

STAR protocols 20210327 2

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- ...[more]

PMID: 33870222

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