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

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.

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Citation

Published Version (Please cite this version)

10.1002/0471140856.tx2011s67

Publication Info

Gonzalez-Hunt, Claudia P, John P Rooney, Ian T Ryde, Charumathi Anbalagan, Rashmi Joglekar and Joel N Meyer (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 https://hdl.handle.net/10161/12422.

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Scholars@Duke

Gonzalez-Hunt

Claudia Gonzalez-Hunt

Research Associate, Senior

Research Interests: Mitochondrial biology, genome integrity, Parkinson's disease, neurodegeneration


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