Quantitative PCR-based measurement of nuclear and mitochondrial DNA damage and repair in mammalian cells.

ABSTRACT: In this chapter, we describe a gene-specific quantitative PCR (QPCR)-based assay for the measurement of DNA damage, using amplification of long DNA targets. This assay has been used extensively to measure the integrity of both nuclear and mitochondrial genomes exposed to different genotoxins and has proven to be particularly valuable in identifying reactive oxygen species-mediated mitochondrial DNA damage. QPCR can be used to quantify both the formation of DNA damage as well as the kinetics of damage removal. One of the main strengths of the assay is that it permits monitoring the integrity of mtDNA directly from total cellular DNA without the need for isolating mitochondria or a separate step of mitochondrial DNA purification. Here we discuss advantages and limitations of using QPCR to as

SUBMITTER: Furda A 

PROVIDER: S-EPMC4407362 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature