Visualization and interpretation of eukaryotic DNA replication intermediates in vivo by electron microscopy
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 uncov...
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| Published in: | Methods in molecular biology (Clifton, N.J.) Vol. 1094; p. 177 |
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| Format: | Journal Article |
| Language: | English |
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United States
01.01.2014
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| ISSN: | 1940-6029, 1940-6029 |
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| 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 microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells. |
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| AbstractList | 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 microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells.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 microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells. 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 microscope. This chapter outlines the procedures required to visualize and interpret in vivo replication intermediates of genomic DNA, extracted from budding yeast, Xenopus egg extracts, or cultured mammalian cells. |
| Author | Lopes, Massimo Neelsen, Kai J Chaudhuri, Arnab Ray Follonier, Cindy Herrador, Raquel |
| Author_xml | – sequence: 1 givenname: Kai J surname: Neelsen fullname: Neelsen, Kai J organization: Institute of Molecular Cancer Research, University of Zurich, Zurich, Switzerland – sequence: 2 givenname: Arnab Ray surname: Chaudhuri fullname: Chaudhuri, Arnab Ray – sequence: 3 givenname: Cindy surname: Follonier fullname: Follonier, Cindy – sequence: 4 givenname: Raquel surname: Herrador fullname: Herrador, Raquel – sequence: 5 givenname: Massimo surname: Lopes fullname: Lopes, Massimo |
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| SubjectTerms | Animals Cell Extracts Chromatin - metabolism Chromosomes, Artificial, Bacterial - metabolism Cross-Linking Reagents - pharmacology DNA - metabolism DNA Replication - drug effects DNA, Cruciform - metabolism Eukaryotic Cells - cytology Eukaryotic Cells - ultrastructure Ficusin - pharmacology Genome, Fungal Male Mammals Microscopy, Electron - methods Nucleic Acid Denaturation - drug effects Saccharomyces cerevisiae - drug effects Saccharomyces cerevisiae - genetics Saccharomyces cerevisiae - metabolism Spermatozoa - cytology Spermatozoa - drug effects Xenopus |
| Title | Visualization and interpretation of eukaryotic DNA replication intermediates in vivo by electron microscopy |
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