DNA methylation detection: bisulfite genomic sequencing analysis.
DNA methylation detection: bisulfite genomic sequencing analysis.
复制标题
DOI：
10.1007/978-1-61779-316-5_2
复制
发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Tollefsbol, Trygve O
中科院分区：
文献类型：
--
作者：
Li, Yuanyuan;Tollefsbol, Trygve O
DNA甲基化通常发生在CpG二核苷酸胞嘧啶的C5位，在基因表达、胚胎发育、细胞增殖、分化和染色体稳定等生物学过程中起着关键作用。异常的DNA甲基化通常与DNA动态平衡的丧失和基因组的不稳定有关，从而导致人类疾病的发展，如癌症。DNA甲基化的重要性使得人们迫切需要高灵敏度和高可靠性的有效方法来探索创新的诊断和治疗策略。Frommer及其同事开发的亚硫酸氢盐基因组测序被认为是基于亚硫酸氢钠转化基因组DNA的DNA甲基化分析的一次革命。除了亚硫酸氢盐基因组测序方法具有定性和定量高等优点外，它还是许多衍生方法的基本原理，以更好地解释DNA甲基化的奥秘。在这里，我们将介绍一种目前在我们实验室中频繁使用的方案，该方案已被证明产生了最佳结果。我们还将讨论该领域中各种应用程序的潜在技术问题和故障排除注意事项。
DNA methylation, which most commonly occurs at the C5 position of cytosines within CpG dinucleotides, plays a pivotal role in many biological procedures such as gene expression, embryonic development, cellular proliferation, differentiation and chromosome stability. Aberrant DNA methylation is often associated with loss of DNA homeostasis and genomic instability leading to development of human diseases such as cancer. The importance of DNA methylation creates an urgent demand for effective methods with highly sensitivity and reliability to explore innovative diagnostic and therapeutic strategies. Bisulfite genomic sequencing developed by Frommer and colleagues was recognized as a revolution in DNA methylation analysis based on conversion of genomic DNA by using sodium bisulfite. Besides various merits of the bisulfite genomic sequencing method such as being highly qualitatively and quantitatively, it serves as a fundamental principle to many derived methods to better interpret the mystery of DNA methylation. Here we will present a protocol currently frequently used in our laboratory that has proved to yield optimal outcomes. We will also discuss the potential technical problems and troubleshooting notes for a variety of applications in this field.