Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis.
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 (10(9)bp) 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.
Sutherland
Betsy M
BM
Biology Department, Brookhaven National Laboratory, Upton, NY 11973-5000, USA.
Georgakilas
Alexandros G
AG
Bennett
Paula V
PV
Laval
Jacques
J
Sutherland
John C
JC
D000818:Animals; D004247:DNA; D004249:DNA Damage / Q000528:radiation effects; D004260:DNA Repair; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005720:Gamma Rays; D006801:Humans; D014741:Video Recording
D004247:DNA