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Use of the DNA Fiber Spreading Technique to Detect the Effects of Mutant p53 on DNA Replication.
MedLine Citation:
PMID:  23150444     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
DNA replication involves a coordinated progression through S phase, and disruption of these regulated steps may cause gene abnormalities, which may lead to cancer. Different stages of DNA replication can be detected immunofluorescently that would indicate how replication is progressing in a cell population or under specific conditions. We describe a method for labeling replicating DNA with two nucleotide analogs, and then detecting the sequential patterns of incorporation using fluorescently labeled antibodies on DNA spread onto a glass slide. Quantification of the different types of replication patterns produced by this method reveals how replication is achieved under different conditions by the predominance and lengths of elongating replication forks progressing from single or clustered origins, as well as the sites of termination from two converging forks.
Authors:
Rebecca A Frum; Sumitra Deb; Swati Palit Deb
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Publication Detail:
Type:  JOURNAL ARTICLE    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  962     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  147-155     Citation Subset:  -    
Affiliation:
Department of Internal Medicine, Massey Cancer Center, Goodwin Research Laboratory, Virginia Commonwealth University, Richmond, VA, USA.
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