Single-sister labeling in the C. elegans germline using the nucleotide analog EdU
Summary: 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 an...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722002246 |
| _version_ | 1817988720144416768 |
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| author | David E. Almanzar Antonia Hamrick Ofer Rog |
| author_facet | David E. Almanzar Antonia Hamrick Ofer Rog |
| author_sort | David E. Almanzar |
| collection | DOAJ |
| description | Summary: 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). |
| first_indexed | 2024-04-14T00:37:46Z |
| format | Article |
| id | doaj.art-73eb4b58d399473ab8ce966840328be0 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-14T00:37:46Z |
| publishDate | 2022-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-73eb4b58d399473ab8ce966840328be02022-12-22T02:22:17ZengElsevierSTAR Protocols2666-16672022-06-0132101344Single-sister labeling in the C. elegans germline using the nucleotide analog EdUDavid E. Almanzar0Antonia Hamrick1Ofer Rog2School of Biological Sciences and Center for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USASchool of Biological Sciences and Center for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USASchool of Biological Sciences and Center for Cell and Genome Sciences, University of Utah, Salt Lake City, UT 84112, USA; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166722002246Cell BiologyGeneticsMicroscopyModel OrganismsMolecular BiologyMolecular/Chemical Probes |
| spellingShingle | David E. Almanzar Antonia Hamrick Ofer Rog Single-sister labeling in the C. elegans germline using the nucleotide analog EdU STAR Protocols Cell Biology Genetics Microscopy Model Organisms Molecular Biology Molecular/Chemical Probes |
| 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 | Cell Biology Genetics Microscopy Model Organisms Molecular Biology Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722002246 |
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