PCR-Based Analysis of Mitochondrial DNA Copy Number, Mitochondrial DNA Damage, and Nuclear DNA Damage.
Curr Protoc Toxicol
; 67: 20.11.1-20.11.25, 2016 Feb 01.
Article
in En
| MEDLINE
| ID: mdl-26828332
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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
DNA Damage
/
DNA, Mitochondrial
/
Cell Nucleus
/
Polymerase Chain Reaction
/
DNA Copy Number Variations
Limits:
Animals
/
Humans
Language:
En
Journal:
Curr Protoc Toxicol
Year:
2016
Document type:
Article