Measurement of DNA damage using agarose gel electrophoresis and electronic imaging
Damage done to DNA by ultraviolet (uv) light, gamma rays and other carcinogens can be quantified using agarose gel electrophororesis. Agents that either produce strand breaks directly or that produce lesions that can be enzymatically or chemically converted to strand breaks can be studied. The method requires: (1) accurate measurement of the disribution of mass of DNA as a function of the distance of migration in the gel, (2) determination of the dispersion function of the electrophoresis system and (3) calculation of weighted averages of these functions by a computer. Less than 50 ng of DNA are required and the DNA need not be labeled with a radioactive tracer. Hence, the damage and repair of DNA in non-dividing cells and intact organisms---including humans---can be studied. Initial applications have focused on the quantitation of cyclobutyl pyrimidine dimers in the DNA of uv irradiated human skin. The sensitivity of lesion detection is increased by unidirectional pulsed field electrophoresis and other methods that separate longer DNA molecules. Replacing photographic detection of ethidium fluorescence by electronic imaging increases the accuracy of the measurement and the speed of data analysis. Quantitative electronic imaging of gel fluorescence offers advantages over photography in other areas of molecular biology, medicine and biotechnology. 26 refs., 5 figs.
- Research Organization:
- Brookhaven National Lab., Upton, NY (USA)
- DOE Contract Number:
- AC02-76CH00016
- OSTI ID:
- 7138962
- Report Number(s):
- BNL-41435; CONF-880777-1; ON: DE88013764
- Resource Relation:
- Conference: 6. meeting of the International Electrophoresis Society, Copenhagen, Denmark, 4 Jul 1988; Other Information: Portions of this document are illegible in microfiche products
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
PYRIMIDINE DIMERS
ELECTROPHORESIS
MAN
OLIGONUCLEOTIDES
SKIN
ULTRAVIOLET RADIATION
ANIMALS
BODY
ELECTROMAGNETIC RADIATION
MAMMALS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
ORGANS
PRIMATES
RADIATIONS
VERTEBRATES
560120* - Radiation Effects on Biochemicals
Cells
& Tissue Culture