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Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo

Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient diges...

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Detalles Bibliográficos
Autores principales: Williams, Ruth M., Sauka-Spengler, Tatjana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8121703/
https://www.ncbi.nlm.nih.gov/pubmed/34027475
http://dx.doi.org/10.1016/j.xpro.2021.100507
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author Williams, Ruth M.
Sauka-Spengler, Tatjana
author_facet Williams, Ruth M.
Sauka-Spengler, Tatjana
author_sort Williams, Ruth M.
collection PubMed
description Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient digestion/ligation type IIs restriction enzyme-based cloning protocol, we develop a multiplexed approach for rapidly identifying enhancer activity. For complete details on the use and execution of this protocol, please see Williams et al. (2019).
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spelling pubmed-81217032021-05-20 Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo Williams, Ruth M. Sauka-Spengler, Tatjana STAR Protoc Protocol Here, we describe a highly efficient, medium-throughput strategy for cloning and in vivo screening of putative enhancers using the chick embryo. By incorporating 48 unique nanotags for use in NanoString nCounter® across three different fluorescent reporters and developing a rapid and efficient digestion/ligation type IIs restriction enzyme-based cloning protocol, we develop a multiplexed approach for rapidly identifying enhancer activity. For complete details on the use and execution of this protocol, please see Williams et al. (2019). Elsevier 2021-05-05 /pmc/articles/PMC8121703/ /pubmed/34027475 http://dx.doi.org/10.1016/j.xpro.2021.100507 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Williams, Ruth M.
Sauka-Spengler, Tatjana
Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_full Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_fullStr Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_full_unstemmed Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_short Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
title_sort rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8121703/
https://www.ncbi.nlm.nih.gov/pubmed/34027475
http://dx.doi.org/10.1016/j.xpro.2021.100507
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