TY - JOUR
T1 - Quantifying clustered DNA damage induction and repair by gel electrophoresis, electronic imaging and number average length analysis
AU - Sutherland, Betsy M.
AU - Georgakilas, Alexandros G.
AU - Bennett, Paula V.
AU - Laval, Jacques
AU - Sutherland, John C.
N1 - Funding Information:
We thank J. Trunk and D. Monteleone (BNL) for hardware and software support in the use of ImageSystem. This work was supported by grants from the Low Dose Program of the Office of Biological and Environmental Research of the US Department of Energy to BMS, from the National Institutes of Health to BMS (R01 CA 86897) and National Space Biomedical Research Institute to A. Gewirtz (University of Pennsylvania) and BMS (IIH00207), by a grant from the National Institutes of Health to JCS (CA 88335) and by CNRS, European Communities (Contract RISC RAD) and Electricité de France, Contrat Radioprotection.
PY - 2003/10/29
Y1 - 2003/10/29
N2 - 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.
AB - 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.
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U2 - 10.1016/j.mrfmmm.2003.08.005
DO - 10.1016/j.mrfmmm.2003.08.005
M3 - Article
C2 - 14637248
AN - SCOPUS:0345016976
SN - 0027-5107
VL - 531
SP - 93
EP - 107
JO - Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
JF - Mutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
IS - 1-2
ER -