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Imaging Replicative Domains in Ultrastructurally Preserved Chromatin by Electron Tomography

Article DOI: 10.3791/62803-v • 14:56 min • May 20th, 2022
May 20th, 2022 •
*1,2, *1,2, *1,3, 1,2, 1, 1, 1, 2, 4, 1,2,5
1Belozersky Institute of Physico-chemical Biology, Lomonosov Moscow State University, 2Faculty of Biology, Lomonosov Moscow State University, 3Faculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, 4National Research Center, Kurchatov Institute, 5V.I. Kulakov National Medical Research Center for Obstetrics, Gynecology, and Perinatology
* These authors contributed equally

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Summary
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This protocol presents a technique for high-resolution mapping of replication sites in structurally preserved chromatin in situ that employs a combination of pre-embedding EdU-streptavidin-Nanogold labeling and ChromEMT.

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Electron Tomography Replicative Domains Ultrastructural Preservation EdU Labeling Silver Enhancement Nanogold Particles ChromEM Staining Glutaraldehyde Fixation Room Temperature Sample Processing Petri Dish Incubator Fixation Time PBS Solution Magnesium Chloride Trione X-100 Plasma Membranes Glycine BSA
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