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. 2018 May;76(1):e50.
doi: 10.1002/cptx.50. Epub 2018 May 18.

Newly Revised Quantitative PCR-Based Assay for Mitochondrial and Nuclear DNA Damage

Laurie H Sanders  1   2 , Jeremy P Rouanet  2 , Evan H Howlett  1 , Tess C Leuthner  3 , John P Rooney  3 , J Timothy Greenamyre  1 , Joel N Meyer  3
Affiliations

Newly Revised Quantitative PCR-Based Assay for Mitochondrial and Nuclear DNA Damage

Laurie H Sanders et al. Curr Protoc Toxicol. 2018 May.

Abstract

Given the crucial role of DNA damage in human health and disease, it is important to be able to accurately measure both mitochondrial and nuclear DNA damage. This article describes a method based on a long-amplicon quantitative PCR-based assay that does not require a separate mitochondrial isolation step, which can often be labor-intensive and generate artifacts. The detailed basic protocol presented here is newly revised, with particular attention to application in Homo sapiens, Rattus norvegicus, and Caenorhabditis elegans resulting from changes in availability of PCR reagents. Optimized extraction support protocols are also described for high-quality DNA from multiple rat tissues for which these procedures had not previously been described. © 2018 by John Wiley & Sons, Inc.

Keywords: Parkinson's disease; QPCR-based assay; mitochondrial DNA damage; neurodegeneration; nuclear DNA damage.

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Conflict of interest statement

CONFLICTS OF INTEREST

The authors declare that there are no conflicts of interest.

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