人端粒长度与线粒体DNA拷贝数双重绝对定量 qPCR 检测试剂盒

Telomeres are repetitive nucleotide elements at the ends of chromosomes that protect chromosomes from degradation and genetic information loss. Normal diploid cells lose telomeres with each cell cycle. Telomere length, therefore, decreases over time and may predict lifespan. Accurate and consistent quantification of telomere length is important in many aspects of cell biology such as chromosomal instability, DNA repair, senescence, apoptosis, cell dysfunctions and oncogenesis. Mitochondrial DNA (mtDNA) is circular, multicopy genome DNA located in mitochondrion, a cellular organelle that plays a key role in the energy production. The capacity for energy production in a cell depends on both mtDNA integrity and copy number. Evidence suggests that telomere length and mitochondrial dysfunction are positively correlated in cellular aging and other age-related disorders such as cancer, diabetes and neurodegenerative diseases. A possible mechanistic link between them has been proposed through the PPAR signaling pathway. However, the specifics of the mechanism still has to be elucidated. ScienCell's Absolute Human Telomere Length and Mitochondrial DNA Copy Number Dual Quantification qPCR Assay Kit (AHDQ) is designed to simultaneously quantify the average telomere length and mtDNA copy number of a human cell population using qPCR. The telomere primer set recognizes and amplifies telomere sequences. The mtDNA primer set recognizes and amplifies one of the most conserved regions on human mtDNA and will not amplify any off-target sequence on nuclear genomic DNA. The single copy reference (SCR) primer set recognizes and amplifies a 100 bp-long region on human chromosome 17, and serves as reference for data normalization. The reference genomic DNA sample with known telomere length and mtDNA copy number serves as a reference for calculating the telomere length and the mtDNA copy number of target samples. The carefully designed primers ensure: (i) high efficiency for trustworthy quantification; and (ii) no non-specific amplification. Each primer set has been validated by qPCR with melt curve analysis and gel electrophoresis for amplification specificity and by template serial dilution for amplification efficiency.

端粒是位于染色体末端的重复核苷酸序列，能够保护染色体免受降解及遗传信息丢失。正常的二倍体细胞在每次细胞周期中都会丢失端粒，因此端粒长度会随时间逐渐缩短，并可作为预测寿命的指标。准确且一致地定量端粒长度在细胞生物学的多个方面都非常重要，例如染色体不稳定性、DNA修复、细胞衰老、凋亡、细胞功能障碍及肿瘤发生等。

线粒体DNA（mtDNA）是一种位于线粒体中的环状、多拷贝基因组DNA。线粒体作为细胞内的重要细胞器，在能量产生过程中发挥关键作用。细胞的能量生成能力取决于mtDNA的完整性及其拷贝数。研究表明，端粒长度与线粒体功能障碍在细胞衰老及其他与年龄相关的疾病（如癌症、糖尿病和神经退行性疾病）中呈正相关。已有研究提出二者可能通过PPAR信号通路存在机制性关联，但具体机制尚待进一步阐明。

ScienCell 人端粒长度与线粒体DNA拷贝数双重绝对定量 qPCR 检测试剂盒（AHDQ）可通过 qPCR 同时定量人群细胞的平均端粒长度和mtDNA拷贝数。端粒引物组可识别并扩增端粒序列；mtDNA引物组可识别人类mtDNA上高度保守的区域并进行扩增，且不会扩增核基因组DNA中的非特异性序列。单拷贝参考基因（SCR）引物组针对人类第17号染色体上一段约100 bp的区域进行扩增，用作数据标准化内参。具有已知端粒长度和mtDNA拷贝数的参考基因组DNA样本可作为标准，用于计算目标样品的端粒长度和mtDNA拷贝数。

精心设计的引物可确保：（i）高扩增效率，从而获得可靠的定量结果；（ii）无非特异性扩增。每组引物均通过qPCR熔解曲线分析和凝胶电泳验证其扩增特异性，并通过模板梯度稀释实验验证其扩增效率。