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Visualization and interpretation of eukaryotic DNA replication intermediates in vivo by electron microscopy


Neelsen, Kai J; Chaudhuri, Arnab Ray; Follonier, Cindy; Herrador, Raquel; Lopes, Massimo (2014). Visualization and interpretation of eukaryotic DNA replication intermediates in vivo by electron microscopy. In: Stockert, Juan C; Espada, Jesús; Blázquez-Castro, Alfonso. Functional Analysis of DNA and Chromatin. New York, USA: Humana Press (Springer), 177-208.

Abstract

The detailed understanding of the DNA replication process requires structural insight. The combination of psoralen cross-linking and electron microscopy has been extensively exploited to reveal the fine architecture of in vivo DNA replication intermediates. This approach proved instrumental to uncover the basic mechanisms of DNA duplication, as well as the perturbation of this process by various forms of replication stress. The replication structures are stabilized in vivo (by psoralen cross-linking) prior to extraction and enrichment procedures, allowing their visualization at the transmission electron micro- scope. This chapter outlines the procedures required to visualize and interpret in vivo replication interme- diates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells.

Abstract

The detailed understanding of the DNA replication process requires structural insight. The combination of psoralen cross-linking and electron microscopy has been extensively exploited to reveal the fine architecture of in vivo DNA replication intermediates. This approach proved instrumental to uncover the basic mechanisms of DNA duplication, as well as the perturbation of this process by various forms of replication stress. The replication structures are stabilized in vivo (by psoralen cross-linking) prior to extraction and enrichment procedures, allowing their visualization at the transmission electron micro- scope. This chapter outlines the procedures required to visualize and interpret in vivo replication interme- diates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells.

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Additional indexing

Item Type:Book Section, not refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Molecular Cancer Research
07 Faculty of Science > Institute of Molecular Cancer Research
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:2014
Deposited On:04 Nov 2013 07:58
Last Modified:05 Apr 2016 17:05
Publisher:Humana Press (Springer)
Series Name:Methods in Molecular Biology
Number:1094
ISSN:1064-3745
ISBN:978-1-62703-705-1
Publisher DOI:https://doi.org/10.1007/978-1-62703-706-8_15
Related URLs:http://link.springer.com/book/10.1007/978-1-62703-706-8

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