“Mirror” Method to Estimate Mutagenic Activity of DNA Lesions
AbstractWe 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
Share & Cite This Article
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.
Gening LV, Shevchenko OV, Kazachenko KY, Tarantul VZ. “Mirror” Method to Estimate Mutagenic Activity of DNA Lesions. Methods and Protocols. 2018; 1(3):32.Chicago/Turabian Style
Gening, Leonid V.; Shevchenko, Oleg V.; Kazachenko, Konstantin Y.; Tarantul, Vyacheslav Z. 2018. "“Mirror” Method to Estimate Mutagenic Activity of DNA Lesions." Methods Protoc. 1, no. 3: 32.
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.