Thielmann H W
Cancer Lett. 1979 May;6(6):311-7. doi: 10.1016/s0304-3835(79)80087-7.
A method for determining methylated purine bases in [3H]N-methyl-N-nitrosourea (MeNOUr) treated DNA is described. The method combines reversed-phase high performance liquid chromatography (HPLC) of methylated DNA after hydrolysis in dilute acid with the determination of radioactivity in the fractionated eluates. The peaks of the respective methylated purines were indentified by the use of internal standards. The method allows quantitative separation of 3-methyl-adenine (m3Ade), 7-methyl-adenine (m7Ade), 3-methyl-guanine (m3Gua), 7-methyl-guanine (m7Gua) and O6-methyl-guanine (m6Gua) within 20 min. Thus the total time required for determination of methylated purines is limited only by radioactivity measurements in the respective fractions.
描述了一种用于测定经[³H]N-甲基-N-亚硝基脲（MeNOUr）处理的DNA中甲基化嘌呤碱基的方法。该方法将稀酸水解后的甲基化DNA进行反相高效液相色谱（HPLC）分析，并结合对分级洗脱液中放射性的测定。通过使用内标物来鉴定各个甲基化嘌呤的峰。该方法能够在20分钟内对3-甲基腺嘌呤（m³Ade）、7-甲基腺嘌呤（m⁷Ade）、3-甲基鸟嘌呤（m³Gua）、7-甲基鸟嘌呤（m⁷Gua）和O⁶-甲基鸟嘌呤（m⁶Gua）进行定量分离。因此，测定甲基化嘌呤所需的总时间仅受各个级分中放射性测量的限制。
