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

Ruth M. Williams^*, Tatjana Sauka-Spengler^*

^*Corresponding author for this work

Division of Developmental Biology & Medicine (L5)

Oxford University

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	100414
Journal	STAR protocols
Volume	2
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2021.100414
Publication status	Published - 18 Jun 2021

Keywords

Genomics
Model Organisms
Molecular Biology
Sequencing
Single Cell

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10.1016/j.xpro.2021.100414

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title = "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.",

keywords = "Genomics, Model Organisms, Molecular Biology, Sequencing, Single Cell",

author = "Williams, {Ruth M.} and Tatjana Sauka-Spengler",

note = "Funding Information: Fluorescence-activated cell sorting was performed at the WIMM Flow Cytometry Facility with Kevin Clark. This work was supported by MRC ( G0902418 ), the Lister Institute for Preventive Medicine Research Prize, John Fell Fund ( 131/038 ), and Leverhulme Trust (grant RPG-2015-026 ) to T.S.-S. Publisher Copyright: {\textcopyright} 2021 The Authors",

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doi = "10.1016/j.xpro.2021.100414",

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AU - Sauka-Spengler, Tatjana

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PY - 2021/6/18

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N2 - 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.

AB - 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.

KW - Genomics

KW - Model Organisms

KW - Molecular Biology

KW - Sequencing

KW - Single Cell

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JO - STAR protocols

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ER -

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

Abstract

Keywords

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