DNA DAMAGE QUANTITATION BY ALKALINE GEL ELECTROPHORESIS.
Physical and chemical agents in the environment, those used in clinical applications, or encountered during recreational exposures to sunlight, induce damages in DNA. Understanding the biological impact of these agents requires quantitation of the levels of such damages in laboratory test systems as well as in field or clinical samples. Alkaline gel electrophoresis provides a sensitive (down to {approx} a few lesions/5Mb), rapid method of direct quantitation of a wide variety of DNA damages in nanogram quantities of non-radioactive DNAs from laboratory, field, or clinical specimens, including higher plants and animals. This method stems from velocity sedimentation studies of DNA populations, and from the simple methods of agarose gel electrophoresis. Our laboratories have developed quantitative agarose gel methods, analytical descriptions of DNA migration during electrophoresis on agarose gels (1-6), and electronic imaging for accurate determinations of DNA mass (7-9). Although all these components improve sensitivity and throughput of large numbers of samples (7,8,10), a simple version using only standard molecular biology equipment allows routine analysis of DNA damages at moderate frequencies. We present here a description of the methods, as well as a brief description of the underlying principles, required for a simplified approach to quantitation of DNA damages by alkaline gel electrophoresis.
- Research Organization:
- BROOKHAVEN NATIONAL LABORATORY (US)
- Sponsoring Organization:
- DOE/OFFICE OF SCIENCE (US)
- DOE Contract Number:
- AC02-98CH10886
- OSTI ID:
- 15008840
- Report Number(s):
- BNL-72423-2004-BC; R&D Project: BO-086 (DOE); KP1102020; TRN: US200427%%406
- Resource Relation:
- Other Information: PBD: 24 Mar 2004; Related Information: DNA REPAIR PROTOCOLS: EUKARYOTIC SYSTEMS, SECOND EDITION
- Country of Publication:
- United States
- Language:
- English
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