Urea improves efficiency of bisulphite-mediated sequencing of 5′-methylcytosine in genomic DNA
Urea improves efficiency of bisulphite-mediated sequencing of 5′-methylcytosine in genomic DNA
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DOI：
10.1093/nar/26.21.5009
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发表时间：
1998-11-01
影响因子：
14.9
通讯作者：
Piper, AA
中科院分区：
文献类型：
--
作者：
Paulin, R;Grigg, GW;Piper, AA
通过亚硫酸氢盐介导的基因组测序方法检测5 '-甲基胞嘧啶在诸如X染色体失活、基因组印记和癌症研究等领域中的作用的研究中具有相当大的帮助。然而，有时在靶DNA区域中获得胞嘧啶至尿嘧啶的完全转化会遇到困难。我们在这里报告了一个简单的改进方法，包括添加尿素的亚硫酸氢盐反应，一个步骤，这大大提高了反应效率，大概是通过保持单链形式的目标DNA，从而允许完全和可靠的转换。
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.