Human Mitochondrial DNA Damage Quantification qPCR Assay Kit

Mitochondrial DNA (mtDNA) is a circular DNA found inside the mitochondria, and it plays a key role in the cells’ energy production. Damage to mtDNA can disrupt cellular energy processes, leading to decreased cell growth and apoptosis. mtDNA damage serves as a meaningful biomarker for assessing the genotoxicity of drugs and environmental toxins. ScienCell's Human Mitochondrial DNA Damage Quantification qPCR Assay Kit (HMDQ) operates on the principle that various DNA lesions can impede DNA polymerase progression. Consequently, DNA with fewer lesions amplifies more readily than damaged DNA under identical conditions. Damage levels can be quantified in terms of the lesions per kilobase pair using a Poisson distribution of lesions or the percentage of intact mtDNA of the target sample to the control sample. Additionally, our assay allows for tracking DNA repair kinetics by measuring the restoration of target DNA amplification over time following the removal of the DNA-damaging agent. This assay monitors the integrity of mtDNA directly from total cellular DNA without the need to isolate mitochondria DNA.
The primer sets (Cat. #8978a and Cat. #8978b) recognizes and amplifies sequences in the most conserved regions on human mtDNA, excluding any off-target sequence on nuclear genomic DNA. We utilize 2X LanaRana Long Range PCR Master Mix (cat #MB6098) and Human Long mtDNA Primer Set (Cat. #8978a) to amplify a long 8.9 kb DNA fragment. For amplifying a short 178 bp mtDNA fragment, we use the 2X GoldNStart TaqGreen qPCR Master Mix (Cat. #MB6018a-1) and Human Short mtDNA Primer Set (Cat. # 8978b). Human DNA from Non-Damaged (untreated) and Damaged (200 µM H 2 O 2 treated) cells serves as positive and negative controls for the reaction.