人类线粒体 DNA 损伤定量 qPCR 检测试剂盒

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.

线粒体 DNA（mtDNA）是存在于线粒体内的环状 DNA，在细胞能量产生中起关键作用。mtDNA 损伤会干扰细胞能量代谢，导致细胞生长减缓和凋亡。mtDNA 损伤可作为评估药物或环境毒素基因毒性的有效生物标志物。ScienCell 的人类线粒体 DNA 损伤定量 qPCR 检测试剂盒（Human Mitochondrial DNA Damage Quantification qPCR Assay Kit，HMDQ）基于这样一个原理：各种 DNA 损伤会阻碍 DNA 聚合酶的延伸。因此，在相同条件下，损伤较少的 DNA 扩增更容易。损伤水平可通过每千碱基对的损伤数（基于泊松分布）或目标样本与对照样本的完整 mtDNA 百分比进行定量。此外，该检测还可通过观察 DNA 损伤因子去除后目标 DNA 扩增恢复情况来追踪 DNA 修复动力学。本检测可直接从总细胞 DNA 中监测 mtDNA 完整性，无需单独分离线粒体 DNA。

试剂盒中的引物组（Cat. #8978a 和 Cat. #8978b）可识别并扩增人类 mtDNA 上最保守区域的序列，同时排除核基因组 DNA 的非特异性序列。我们使用 2X LanaRana Long Range PCR Master Mix（Cat. #MB6098）和 Human Long mtDNA Primer Set（Cat. #8978a）扩增 8.9 kb 的长片段 DNA；而扩增短片段（178 bp）mtDNA 时，则使用 2X GoldNStart TaqGreen qPCR Master Mix（Cat. #MB6018a-1）和 Human Short mtDNA Primer Set（Cat. #8978b）。来自未处理（Non-Damaged）和 H₂O₂ 处理（200 µM Damaged）细胞的 DNA 分别作为反应的阳性和阴性对照。