人 ELOVL2 启动子及全基因组DNA甲基化双重定量 qPCR 检测试剂盒

DNA methylation at CpG islands in promoters regulates gene expression. It is a dynamic epigenetic process affected by various genetic and environmental factors. Numerous diseases including cancer have been linked to aberrant global changes in genomic DNA methylation. It is generally thought that global genomic DNA methylation level can be represented by the methylation level at repetitive elements. One example of this includes the repetitive Long Interspersed Nuclear Element-1 (LINE-1), which compromises 17-18% of the human genome and is usually heavily methylated in most normal primary cells. Furthermore, the CpG island methylation level at human ELOVL2 gene promoter has been well-associated with chronological age in various populations, cell types, and tissues. ELOVL2 promoter methylation level is therefore a potential marker for human chronological age. ScienCell's Human ELOVL2 Promoter and Global DNA Dual Methylation Quantification qPCR Assay Kit (HEGDD) is designed to simultaneously quantify the level of human ELOVL2 promoter and LINE1 promoter methylation. The reference DNA sample consists of a 1:1 ratio of methylated ELOVL2 promoter copies to non-methylated ELOVL2 promotor copies (both copies represent bisulfite converted promoter sequences) and a 1:1 ratio of methylated LINE1 promoter copies to non-methylated LINE1 promotor copies (both copies represent bisulfite converted promoter sequences), and serves as a reference for calculating the ratio of methylated to non-methylated ELOVL2 promotor and LINE1 promoter of target samples. The carefully designed primers ensure: (i) high efficiency for trustworthy quantification; and (ii) negligible 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.

DNA 甲基化（发生于启动子区 CpG 岛）在基因表达调控中发挥重要作用。这是一种动态的表观遗传过程，会受到多种遗传和环境因素的影响。包括癌症在内的多种疾病均与全基因组DNA甲基化异常变化密切相关。通常认为，全基因组DNA甲基化水平可以通过重复序列的甲基化水平来反映。其中一个典型代表是长散在核元件-1（LINE-1），其约占人类基因组的17%–18%，并且在大多数正常初代细胞中通常处于高度甲基化状态。此外，人类 ELOVL2 基因启动子区域的 CpG 岛甲基化水平在不同人群、细胞类型和组织中均与实际年龄高度相关，因此可作为评估人类实际年龄的潜在生物标志物。

ScienCell 人 ELOVL2 启动子及全基因组DNA甲基化双重定量 qPCR 检测试剂盒（HEGDD）可同时定量检测人类 ELOVL2 启动子和 LINE1 启动子的甲基化水平。该试剂盒中的参考DNA样本包括：甲基化与非甲基化的 ELOVL2 启动子拷贝按 1:1 比例组成（均为经亚硫酸氢盐转化后的启动子序列），以及甲基化与非甲基化的 LINE1 启动子拷贝按 1:1 比例组成（同样为经亚硫酸氢盐转化后的启动子序列），可作为标准用于计算目标样品中 ELOVL2 和 LINE1 启动子的甲基化与非甲基化比例。

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）极低的非特异性扩增。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。