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Estimation of the Mutagenic Potential of 8-Oxog in Nuclear Extracts of Mouse Cells Using the “Framed Mirror” Method

Institute of Molecular Genetics, Kurchatov Sq. 2, Moscow 123182, Russia
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Methods Protoc. 2020, 3(1), 3; https://doi.org/10.3390/mps3010003
Received: 11 November 2019 / Revised: 24 December 2019 / Accepted: 31 December 2019 / Published: 3 January 2020
We propose an improved earlier described “mirror” method for detecting in cell nuclear extracts mutations that arise in DNA during its replication due to the misincorporation of deoxyadenosine-5′-monophosphate (dAMP) opposite 7,8-dihydro-8-oxoguanine (8-oxoG). This method is based on the synthesis of a complementary chain (“mirror”) by nuclear extracts of different mice organs on a template containing 8-oxoG and dideoxycytidine residue (ddC) at the 3′‑end. The “mirror” was amplified by PCR using primers part of which was non-complementary to the template. It allowed obtaining the “framed mirror” products. The misincorporation of dAMP in “framed mirror” products forms an EcoRI restriction site. The restriction analysis of double-stranded “framed mirror” products allows a quantification of the mutation frequency in nuclear extracts. The data obtained show that the mutagenic potential of 8-oxoG markedly varied in different organs of adult mice and embryos. View Full-Text
Keywords: DNA lesions; 7,8-dihydro-8-oxoguanine (8-oxoG); mutagenic activity; method of detection; cell nuclear extracts; mice organs and embryos DNA lesions; 7,8-dihydro-8-oxoguanine (8-oxoG); mutagenic activity; method of detection; cell nuclear extracts; mice organs and embryos
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MDPI and ACS Style

Gening, L.V.; Volodin, A.A.; Kazachenko, K.Y.; Makarova, I.V.; Tarantul, V.Z. Estimation of the Mutagenic Potential of 8-Oxog in Nuclear Extracts of Mouse Cells Using the “Framed Mirror” Method. Methods Protoc. 2020, 3, 3. https://doi.org/10.3390/mps3010003

AMA Style

Gening LV, Volodin AA, Kazachenko KY, Makarova IV, Tarantul VZ. Estimation of the Mutagenic Potential of 8-Oxog in Nuclear Extracts of Mouse Cells Using the “Framed Mirror” Method. Methods and Protocols. 2020; 3(1):3. https://doi.org/10.3390/mps3010003

Chicago/Turabian Style

Gening, Leonid V., Alexandr A. Volodin, Konstantin Y. Kazachenko, Irina V. Makarova, and Vyacheslav Z. Tarantul 2020. "Estimation of the Mutagenic Potential of 8-Oxog in Nuclear Extracts of Mouse Cells Using the “Framed Mirror” Method" Methods and Protocols 3, no. 1: 3. https://doi.org/10.3390/mps3010003

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