尿素可提高亚硫酸氢盐介导的基因组DNA中5'-甲基胞嘧啶测序的效率。
Urea improves efficiency of bisulphite-mediated sequencing of 5'-methylcytosine in genomic DNA.
作者信息
Paulin R, Grigg G W, Davey M W, Piper A A
机构信息
Department of Cell and Molecular Biology, University of Technology, Sydney, Westbourne Street, Gore Hill, NSW 2065, Australia.
出版信息
Nucleic Acids Res. 1998 Nov 1;26(21):5009-10. doi: 10.1093/nar/26.21.5009.
The detection of 5'-methylcytosine by the bisulphite-mediated genomic sequencing method has considerably aided study of the role of methylation in areas such as X chromosome inactivation, genomic imprinting and cancer research. However on occasion difficulty has been experienced in obtaining complete conversion of cytosine to uracil in regions of the target DNA. We report here a simple improvement to the method involving addition of urea to the bisulphite reaction, a step which greatly improves the reaction efficiency, presumably by maintaining the target DNA in single stranded form, thereby allowing complete and reliable conversion.
通过亚硫酸氢盐介导的基因组测序方法检测5'-甲基胞嘧啶，极大地促进了甲基化在X染色体失活、基因组印记和癌症研究等领域作用的研究。然而，有时在目标DNA区域很难实现胞嘧啶完全转化为尿嘧啶。我们在此报告一种对该方法的简单改进，即在亚硫酸氢盐反应中添加尿素，这一步骤极大地提高了反应效率，推测是通过将目标DNA保持为单链形式，从而实现完全且可靠的转化。
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