Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform.
Accurate quantification of DNA methylation using combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform.
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DOI：
10.1093/nar/gnj017
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发表时间：
2006-02-07
影响因子：
14.9
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中科院分区：
文献类型：
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DNA 甲基化是研究最充分的表观遗传修饰，描述了胞嘧啶向 5-甲基胞嘧啶的转化。这种现象的重要性在于，异常启动子高甲基化在癌症中很常见，并且经常与基因沉默相关。目前有多种技术可用于 DNA 甲基化分析。然而，准确且可重复的 DNA 甲基化定量仍然具有挑战性。在本报告中，我们将 Bio-COBRA（亚硫酸氢盐限制性酶切分析与 Agilent 2100 生物分析仪平台相结合）描述为一种定量 DNA 甲基化分析的新方法。 COBRA 是一种成熟的方法，通过对 PCR 扩增的亚硫酸氢盐处理的 DNA 进行限制性内切酶分析来询问 DNA 甲基化，与生物分析仪平台相结合，可以快速定量评估大样本组中的 DNA 甲基化模式。 Bio-COBRA 的灵敏度和重现性使其成为分析临床样本中 DNA 甲基化的宝贵工具，有助于开发疾病检测和管理方面的诊断和预后参数。
DNA methylation is the best-studied epigenetic modification and describes the conversion of cytosine to 5-methylcytosine. The importance of this phenomenon is that aberrant promoter hypermethylation is a common occurrence in cancer and is frequently associated with gene silencing. Various techniques are currently available for the analysis of DNA methylation. However, accurate and reproducible quantification of DNA methylation remains challenging. In this report, we describe Bio-COBRA (combined bisulfite restriction analysis coupled with the Agilent 2100 Bioanalyzer platform), as a novel approach to quantitative DNA methylation analysis. The combination of a well-established method, COBRA, which interrogates DNA methylation via the restriction enzyme analysis of PCR-amplified bisulfite treated DNAs, with the Bioanalyzer platform allows for the rapid and quantitative assessment of DNA methylation patterns in large sample sets. The sensitivity and reproducibility of Bio-COBRA make it a valuable tool for the analysis of DNA methylation in clinical samples, which could aid in the development of diagnostic and prognostic parameters with respect to disease detection and management.