Mitochondrial dna damage assay

Mitochondrial Dna Damage Assay. Mitochondrial DNA mtDNA is highly susceptible to oxidative damage. Certainly at least one chemotherapeutic regimen CHOP therapy that includes doxorubicin can induce cardiomyopathy through mitochondrial genotoxicity in cardiac muscle cells. Mitochondrial function was monitored using the MTT 3 45-dimethylthiazol-2-yl25-diphenyltetrazolium bromide assay. We identified the molecular identity of mtDNA damage.

Here we describe methods for quantifying oxidative mtDNA damage and mutagenesis in S. DNA damage in the mitochondrial genome was measured utilizing a PCR-based assay currently the most robust way of measuring damage in mtDNA 57. This assay has been extensively used to measure the integrity of both nuclear and mitochondrial genomes exposed to different genotoxins and has proved particularly valuable in identifying reactive oxygen species-mediated mitochondrial DNA mtDNA 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 PCR amplification of a large mtDNA segment 89 kb a small mtDNA segment 221 bp and the gene of polymerase β 122 kb was performed. 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 the formation of DNA damage as well as the kinetics of damage removal.

DNA damage can cause and result from oxidative stress and mitochondrial impairment both of which are implicated in the pathogenesis of Parkinsons disease PD. Mitochondrial DNA mtDNA is highly susceptible to oxidative damage. DNA damage was documented as increased levels of γH2AX pCHK2 by Comet assay AIM2 induction and by increased DNA repair non-homologous end joining signaling. 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. The PCR amplification of a large mtDNA segment 89 kb a small mtDNA segment 221 bp and the gene of polymerase β 122 kb was performed. Cerevisiae several of which.