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Methods Protoc. 2018, 1(3), 32; https://doi.org/10.3390/mps1030032

“Mirror” Method to Estimate Mutagenic Activity of DNA Lesions

Institute of Molecular Genetics, Kurchatov Sq. 2, Moscow 123182, Russia
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Received: 29 June 2018 / Revised: 16 August 2018 / Accepted: 24 August 2018 / Published: 27 August 2018
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Abstract

We propose an improved method for detecting mutations that arise in DNA due to misincorporations of deoxyadenosine-5′-monophosphate (dAMP) opposite 7,8-dihydro-8-oxoguanine (8-oxoG). The method is based on the synthesis of complementary chains (“mirror” products) using a template containing 8-oxoG. The misincorporation of dAMP in the “mirror” product forms EcoRI sites. The restriction analysis of double-stranded DNAs obtained by PCR of “mirror” product allows quantification of the mutagenesis frequency. In addition, single-strand conformational polymorphism (SSCP) analysis of the single-stranded “mirror” products shows that different DNA polymerases only incorporate dA or dC opposite 8-oxoG. The proposed approach used in developing this technique can be applied in the study of other lesions as well, both single and clustered. View Full-Text
Keywords: DNA lesions; 7,8-dihydro-8-oxoguanine; 8-oxoG; mutagenic activity; method of detection DNA lesions; 7,8-dihydro-8-oxoguanine; 8-oxoG; mutagenic activity; method of detection
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Gening, L.V.; Shevchenko, O.V.; Kazachenko, K.Y.; Tarantul, V.Z. “Mirror” Method to Estimate Mutagenic Activity of DNA Lesions. Methods Protoc. 2018, 1, 32.

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