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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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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). |
format | Online Article Text |
id | pubmed-9059099 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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