PCR-Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage.

Anbalagan, C; Gonzalez-Hunt, CP; Joglekar, R; Meyer, Joel; Rooney, JP; Ryde, IT

doi:10.1002/0471140856.tx2011s67

Abstract

Because of the role that DNA damage and depletion play in human disease, it is important to develop and improve tools to assess these endpoints. This unit describes PCR-based methods to measure nuclear and mitochondrial DNA damage and copy number. Long amplicon quantitative polymerase chain reaction (LA-QPCR) is used to detect DNA damage by measuring the number of polymerase-inhibiting lesions present based on the amount of PCR amplification; real-time PCR (RT-PCR) is used to calculate genome content. In this unit, we provide step-by-step instructions to perform these assays in Homo sapiens, Mus musculus, Rattus norvegicus, Caenorhabditis elegans, Drosophila melanogaster, Danio rerio, Oryzias latipes, Fundulus grandis, and Fundulus heteroclitus, and discuss the advantages and disadvantages of these assays.

Publication Info

Anbalagan, C; Gonzalez-Hunt, CP; Joglekar, R; Meyer, Joel; Rooney, JP; & Ryde, IT (2016). PCR-Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage. Curr Protoc Toxicol, 67. pp. 20.11.1-20.11.25. 10.1002/0471140856.tx2011s67. Retrieved from http://hdl.handle.net/10161/12422.

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Scholars@Duke

Rashmi Joglekar

Student

Joel Meyer

Truman and Nellie Semans/Alex Brown and Sons Associate Professor of Molecular Environmental Toxicology

Dr. Meyer studies the effects of toxic agents and stressors on human and wildlife health. He is particularly interested in understanding the mechanisms by which environmental agents cause DNA damage, the molecular processes that organisms employ to protect prevent and repair DNA damage, and genetic differences that may lead to increased or decreased sensitivity to DNA damage. Mitochondrial DNA damage and repair, as well as mitochondrial function in general, are a particular focus. He studies

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