Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis

Betsy M. Sutherland, Alexandros G. Georgakilas, Paula V. Bennett, Jacques Laval, John C. Sutherland

Research output: Contribution to journalArticle

61 Scopus citations

Abstract

Assessing DNA damage induction, repair and consequences of such damages requires measurement of specific DNA lesions by methods that are independent of biological responses to such lesions. Lesions affecting one DNA strand (altered bases, abasic sites, single strand breaks (SSB)) as well as damages affecting both strands (clustered damages, double strand breaks) can be quantified by direct measurement of DNA using gel electrophoresis, gel imaging and number average length analysis. Damage frequencies as low as a few sites per gigabase pair (109bp) can be quantified by this approach in about 50ng of non-radioactive DNA, and single molecule methods may allow such measurements in DNA from single cells. This review presents the theoretical basis, biochemical requirements and practical aspects of this approach, and shows examples of their applications in identification and quantitation of complex clustered damages.

Original languageEnglish (US)
Pages (from-to)93-107
Number of pages15
JournalMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
Volume531
Issue number1-2
DOIs
Publication statusPublished - Oct 29 2003
Externally publishedYes

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ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

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