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

STAR protocols (2021-04-20)
Ruth M Williams, Tatjana Sauka-Spengler
RÉSUMÉ

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).

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Sigma-Aldrich
Albumine de sérum bovin, heat shock fraction, protease free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Collagénase from Clostridium histolyticum, Sigma Blend Type L, ≤1.0 FALGPA units/mg solid