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High-resolution mapping of points of site-specific replication stalling
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Vengrova, Sonya and Dalgaard, Jacob Z. (2009) High-resolution mapping of points of site-specific replication stalling. In: Vengrova, Sonya and Dalgaard, Jacob Z., (eds.) DNA replication : methods and protocols. Methods in molecular biology, Vol.521 . New York: Humana Press, pp. 215-227. ISBN 9781603278140
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Official URL: http://dx.doi.org/10.1007/978-1-60327-815-7_12
Abstract
Genetic instability due to stalled replication forks is thought to underlie a number of human diseases, such as premature ageing and cancer susceptibility syndromes. In addition, site-specific stalling occurs at some genetic loci. A detailed understanding of the topology of the stalled replication fork gives a valuable insight into the causes and mechanisms of replication stalling. The method described here allows mapping of the position of the 3′-end of the nascent leading or lagging strand at the replication fork, stalled at a site-specific barrier. The replicating DNA is purified, digested with restriction enzymes, and enriched by BND-cellulose chromatography. The DNA is separated on a sequencing gel, transferred to a membrane, and hybridised to a strand-specific probe. The data obtained using this method allow determining the position of the 3′-end of the nascent strand at a stalled fork with a one-nucleotide resolution.
| Item Type: | Book Item |
|---|---|
| Subjects: | R Medicine > R Medicine (General) |
| Divisions: | Faculty of Medicine > Warwick Medical School > Biomedical Cell Biology Faculty of Medicine > Warwick Medical School |
| Series Name: | Methods in molecular biology |
| Journal or Publication Title: | Methods in Molecular Biology (Clifton, N.J.) |
| Publisher: | Humana Press |
| Place of Publication: | New York |
| ISBN: | 9781603278140 |
| ISSN: | 1064-3745 |
| Book Title: | DNA replication : methods and protocols |
| Editor: | Vengrova, Sonya and Dalgaard, Jacob Z. |
| Date: | 23 January 2009 |
| Volume: | Vol.521 |
| Number of Pages: | 0 |
| Page Range: | pp. 215-227 |
| Identification Number: | 10.1007/978-1-60327-815-7_12 |
| Status: | Peer Reviewed |
| Publication Status: | Published |
| Access rights to Published version: | Restricted or Subscription Access |
| URI: | http://wrap.warwick.ac.uk/id/eprint/45836 |
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