PUBLICATION
The QPCR assay for analysis of mitochondrial DNA damage, repair, and relative copy number
- Authors
- Hunter, S.E., Jung, D., Di Giulio, R.T., and Meyer, J.N.
- ID
- ZDB-PUB-100211-6
- Date
- 2010
- Source
- Methods (San Diego, Calif.) 51(4): 444-451 (Journal)
- Registered Authors
- Di Giulio, Richard T.
- Keywords
- Quantitative PCR assay, Mitochondrial DNA damage, Mitochondrial DNA repair
- MeSH Terms
-
- Animals
- Base Sequence
- DNA Damage*
- DNA Primers/genetics
- DNA Repair*
- DNA, Mitochondrial/analysis*
- DNA, Mitochondrial/genetics*
- DNA, Mitochondrial/metabolism
- Gene Dosage
- Genome, Mitochondrial
- Humans
- Mitochondria/genetics
- Mitochondria/metabolism
- Polymerase Chain Reaction/methods*
- PubMed
- 20123023 Full text @ Methods
Citation
Hunter, S.E., Jung, D., Di Giulio, R.T., and Meyer, J.N. (2010) The QPCR assay for analysis of mitochondrial DNA damage, repair, and relative copy number. Methods (San Diego, Calif.). 51(4):444-451.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping