Journal article
Improved method for measuring the ensemble average of strand breaks in genomic DNA
Environmental and molecular mutagenesis, Vol.38(2‐3), pp.166-174
2001
Handle:
https://hdl.handle.net/2376/113793
PMID: 11746751
Abstract
The cis‐syn cyclobutane pyrimidine dimer (CPD) is the major photoproduct induced in DNA by low wavelength ultraviolet radiation. An improved method was developed to detect CPD formation and removal in genomic DNA that avoids the problems encountered with the standard method of endonuclease detection of these photoproducts. Since CPD‐specific endonucleases make single‐strand cuts at CPD sites, quantification of the frequency of CPDs in DNA is usually done by denaturing gel electrophoresis. The standard method of ethidium bromide staining and gel photography requires more than 10 μg of DNA per gel lane, and correction of the photographic signal for the nonlinear film response. To simplify this procedure, a standard Southern blot protocol, coupled with phosphorimage analysis, was developed. This method uses random hybridization probes to detect genomic sequences with minimal sequence bias. Because of the vast linearity range of phosphorimage detection, scans of the signal profiles for the heterogeneous population of DNA fragments can be integrated directly to determine the number‐average size of the population. Environ. Mol. Mutagen. 38:166–174, 2001 © 2001 Wiley‐Liss, Inc.
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Details
- Title
- Improved method for measuring the ensemble average of strand breaks in genomic DNA
- Creators
- Vyacheslav A BespalovAntonio ConconiXiaoyi ZhangDeirdre FahyMichael J Smerdon
- Publication Details
- Environmental and molecular mutagenesis, Vol.38(2‐3), pp.166-174
- Academic Unit
- Molecular Biosciences, School of
- Publisher
- John Wiley & Sons, Inc; New York
- Number of pages
- 9
- Grant note
- National Institute of Environmental Health Sciences (ES04106; ES02614)
- Identifiers
- 99900548235201842
- Language
- English
- Resource Type
- Journal article