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Single-sister labeling in the C. elegans germline using the nucleotide analog EdU

Reciprocal exchanges between genetically identical sister chromatids (sister chromatid exchanges or SCEs) have been challenging to study. Here, we describe a protocol that utilizes a pulse/chase of the thymidine analog 5-ethyl-3′-deoxyuridine (EdU) in combination with click chemistry and antibody la...

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Detalles Bibliográficos
Autores principales: Almanzar, David E., Hamrick, Antonia, Rog, Ofer
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9059099/
https://www.ncbi.nlm.nih.gov/pubmed/35509971
http://dx.doi.org/10.1016/j.xpro.2022.101344
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author Almanzar, David E.
Hamrick, Antonia
Rog, Ofer
author_facet Almanzar, David E.
Hamrick, Antonia
Rog, Ofer
author_sort Almanzar, David E.
collection PubMed
description Reciprocal exchanges between genetically identical sister chromatids (sister chromatid exchanges or SCEs) have been challenging to study. Here, we describe a protocol that utilizes a pulse/chase of the thymidine analog 5-ethyl-3′-deoxyuridine (EdU) in combination with click chemistry and antibody labeling to selectively label sister chromatids in the C. elegans germline. Labeling has no discernable effects on meiosis, allowing for cytological quantification of SCEs. This protocol can be combined with a variety of imaging approaches, including STED, confocal and super-resolution. For complete details on the use and execution of this protocol, please refer to Almanzar et al. (2021).
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spelling pubmed-90590992022-05-03 Single-sister labeling in the C. elegans germline using the nucleotide analog EdU Almanzar, David E. Hamrick, Antonia Rog, Ofer STAR Protoc Protocol Reciprocal exchanges between genetically identical sister chromatids (sister chromatid exchanges or SCEs) have been challenging to study. Here, we describe a protocol that utilizes a pulse/chase of the thymidine analog 5-ethyl-3′-deoxyuridine (EdU) in combination with click chemistry and antibody labeling to selectively label sister chromatids in the C. elegans germline. Labeling has no discernable effects on meiosis, allowing for cytological quantification of SCEs. This protocol can be combined with a variety of imaging approaches, including STED, confocal and super-resolution. For complete details on the use and execution of this protocol, please refer to Almanzar et al. (2021). Elsevier 2022-04-23 /pmc/articles/PMC9059099/ /pubmed/35509971 http://dx.doi.org/10.1016/j.xpro.2022.101344 Text en © 2022 The Author(s) 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
Almanzar, David E.
Hamrick, Antonia
Rog, Ofer
Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title_full Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title_fullStr Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title_full_unstemmed Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title_short Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
title_sort single-sister labeling in the c. elegans germline using the nucleotide analog edu
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9059099/
https://www.ncbi.nlm.nih.gov/pubmed/35509971
http://dx.doi.org/10.1016/j.xpro.2022.101344
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