亚硫酸氢盐测序法检测小鼠胚胎Oct4启动子区甲基化

应用亚硫酸氢盐测序技术,以昆明小白鼠几种不同时期体内胚为研究对象,对其单拷贝基因Oct4启动子区进行甲基化检测.基因组经低熔点琼脂糖包埋、亚硫酸氢盐修饰后,运用巢式PCR对处理后的DNA进行两轮扩增,扩增产物通过琼脂糖凝胶电泳检测和单克隆测序进行分析.结果表明,亚硫酸氢盐测序法是一项敏感而有效的DNA甲基化检测手段.

DNA methylation detection of the Oct4 promoter in mouse embryos by the bisulfite sequencing method

In this experiment,bisulfite-sequencing technology was used to detect the methylation pattern of the promoter region of Oct4,a single copy gene at several stages during embryonic development of the Kunming White mouse.The genome was embedded in low melting point(LMP) agrose,then modified with bisulfite.After double amplifications of nested PCR,the products were analysed by agarose gel electrophoresis and individual clone sequencing.The result demonstrated that the bisulfite sequencing is a sensitive and effective method to detect DNA methylation.