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.
Type
Journal articleSubject
C. elegansDNA damage
mitochondrial DNA
mitochondrial DNA copy number
qPCR
Animals
Cell Nucleus
DNA Copy Number Variations
DNA Damage
DNA Mutational Analysis
DNA Primers
DNA, Mitochondrial
Humans
Polymerase Chain Reaction
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https://hdl.handle.net/10161/12422Published Version (Please cite this version)
10.1002/0471140856.tx2011s67Publication Info
Gonzalez-Hunt, Claudia P; Rooney, John P; Ryde, Ian T; Anbalagan, Charumathi; Joglekar,
Rashmi; & Meyer, Joel N (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.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Claudia Gonzalez-Hunt
Research Associate, Senior
Research Interests: Mitochondrial biology, genome integrity, Parkinson's disease,
neurodegeneration
Joel Meyer
Associate Professor of Environmental Genomics in the Division of Environmental Sciences
and Policy
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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