人类核 DNA 损伤定量 qPCR 检测试剂盒

Damage to nuclear DNA (nucDNA) is widely considered a key factor in the development of cancer, neurodegenerative disorders, mitochondrial dysfunction, and various age-related conditions. nucDNA damage serves as a meaningful biomarker for assessing the genotoxicity of drugs and environmental toxins. ScienCell's Human Nuclear DNA Damage Quantification qPCR Assay Kit (HNDQ) 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 nucDNA 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 nucDNA.

The primer sets (Cat. #9008a and Cat. #9008b) recognizes and amplifies sequences in the most conserved regions on human nucDNA. We utilize 2X LanaRana Long Range PCR Master Mix (cat #MB6098) and Human Long nucDNA Primer Set (Cat. #9008a) to amplify a long 8.1 kb DNA fragment. For amplifying a short 151 bp nucDNA fragment, we use the 2X GoldNStart TaqGreen qPCR Master Mix (Cat. #MB6018a-1) and Human Short nucDNA Primer Set (Cat. # 9008b). Human DNA from Non-Damaged (untreated) and Damaged (UV treated) cells serves as positive and negative controls for the reaction.

核 DNA（nucDNA）损伤被广泛认为是癌症、神经退行性疾病、线粒体功能障碍以及多种与年龄相关疾病发生的关键因素。核 DNA 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 的人类核 DNA 损伤定量 qPCR 检测试剂盒（Human Nuclear DNA Damage Quantification qPCR Assay Kit，HNDQ）基于这样的原理：各种 DNA 损伤会阻碍 DNA 聚合酶延伸，从而影响扩增效率。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核 DNA 百分比进行定量。此外，该检测可通过观察 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。本检测可直接监测核 DNA 的完整性。

试剂盒中的引物组（Cat. #9008a 和 Cat. #9008b）可识别人类核 DNA 上最保守区域的序列并进行扩增。我们使用 2X LanaRana Long Range PCR Master Mix（Cat. #MB6098）和 Human Long nucDNA Primer Set（Cat. #9008a）扩增 8.1 kb 的长片段 DNA；而扩增 151 bp 短片段时，则使用 2X GoldNStart TaqGreen qPCR Master Mix（Cat. #MB6018a-1）和 Human Short nucDNA Primer Set（Cat. #9008b）。来自未处理（Non-Damaged）和 UV 处理（Damaged）细胞的 DNA 分别作为反应的阳性和阴性对照。