Quantitation of single- and double-strand DNA breaks in vitro and in vivo
This communication describes a rapid and convenient procedure for quantitation of strand breaks in bacterial DNA, both in vitro and in vivo, using agarose gel electrophoresis. The electrophoretic determination of single strand breaks is carried out in alkaline medium, followed by renaturation of the gel and intercalation of the fluorescent dye, ethidium bromide. Double-strand breaks are determined by electrophoresis in neutral medium containing the dye. The distribution of DNA fragment sizes, the determination of the number-average molecular weight, the quantitation of the average number of DNA breaks per molecule, and the ratio between the single- and double-strand breaks are evaluated from microdensitometric scanning of the gels. The application of this analysis to damage caused by a combination of ascorbate and copper is demonstrated.
- OSTI ID:
- 5118582
- Journal Information:
- Anal. Biochem.; (United States), Vol. 2
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ASCORBIC ACID
BIOLOGICAL EFFECTS
COPPER
DNA
ELECTROPHORESIS
STRAND BREAKS
ESCHERICHIA COLI
MEASURING METHODS
BACTERIA
ELEMENTS
METALS
MICROORGANISMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
TRANSITION ELEMENTS
VITAMINS
560302* - Chemicals Metabolism & Toxicology- Microorganisms- (-1987)