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Single-sister Labeling in the C. Elegans Germline Using the Nucleotide Analog EdU.

STAR protocols(2022)

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摘要
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-30-deoxyur-idine (EdU) in combination with click chemistry and antibody labeling to selec-tively label sister chromatids in the C. elegans germline. Labeling has no discern-able 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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关键词
Cell Biology,Genetics,Microscopy,Model Organisms,Molecular Biology,Molecular/Chemical Probes
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