The QPCR assay for analysis of mitochondrial DNA damage, repair, and relative copy number.

Published

Journal Article (Review)

The quantitative polymerase chain reaction (QPCR) assay allows measurement of DNA damage in the mitochondrial and nuclear genomes without isolation of mitochondria. It also permits measurement of relative mitochondrial genome copy number. Finally, it can be used for measurement of DNA repair in vivo when employed appropriately. In this manuscript we briefly review the methodology of the QPCR assay, discuss its strengths and limitations, address considerations for measurement of mitochondrial DNA repair, and describe methodological changes implemented in recent years. We present QPCR assay primers and reaction conditions for five species not previously described in a methods article: Caenorhabditis elegans, Fundulus heteroclitus, Danio rerio, Drosophila melanogaster, and adenovirus. Finally, we illustrate the use of the assay by measuring repair of ultraviolet C radiation-induced DNA damage in the nuclear but not mitochondrial genomes of a zebrafish cell culture.

Full Text

Duke Authors

Cited Authors

  • Hunter, SE; Jung, D; Di Giulio, RT; Meyer, JN

Published Date

  • August 2010

Published In

Volume / Issue

  • 51 / 4

Start / End Page

  • 444 - 451

PubMed ID

  • 20123023

Pubmed Central ID

  • 20123023

Electronic International Standard Serial Number (EISSN)

  • 1095-9130

International Standard Serial Number (ISSN)

  • 1046-2023

Digital Object Identifier (DOI)

  • 10.1016/j.ymeth.2010.01.033

Language

  • eng