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Bisulte-mediated Deamination of Cytosine in DNA under Near-neutral Conditions

机译：在近中性条件下双硫醚介导的DNA胞嘧啶脱氨

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In DNA methylation analysis for epigenetic studies, the most frequently used procedure involves bisulfite-mediated conversion of cytosine to uracil. Resistance of 5- methylcytosine against this bisulfite reaction is the basis for identifying the methylated sites. This chemical modifi-cation consists of a treatment of single-stranded nucleic acids with high-concentration bisufite at pH 5–6. We report here that the treatment can be performed at pHa6 by supplementary presence of tetramethylammonium ion. Quaternary ammonium cation appears to be essential in this unique enhancement of the reaction in the near-neutral pH range, as betaine was also efiective but trimethylamine was not. Using a tetramethylammonium-bisulfite reagent at pH 6.4, we achieved a complete deamination of cytosine residues in a DNA molecule, without any significant changes taking place for 5-methylcytosine in it. These findings have opened up the possibility of improving and expanding the DNA methylation analysis technology.
机译：在用于表观遗传学研究的DNA甲基化分析中，最常用的方法涉及亚硫酸氢盐介导的胞嘧啶向尿嘧啶的转化。 5-甲基胞嘧啶对该亚硫酸氢盐反应的抗性是鉴定甲基化位点的基础。这种化学修饰包括在pH值为5-6的情况下用高浓度的亚硫酸氢盐处理单链核酸。我们在这里报告说，可通过补充存在四甲基铵离子在pHa6下进行治疗。在接近中性的pH范围内，季铵阳离子似乎对这种独特的反应增强至关重要，因为甜菜碱也是有效的，而三甲胺不是。使用pH 6.4的四甲基亚硫酸氢盐试剂，我们可以完全脱去DNA分子中的胞嘧啶残基，而其中的5-甲基胞嘧啶则没有任何重大变化。这些发现为改进和扩展DNA甲基化分析技术开辟了可能性。

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