Rein T, DePamphilis M L, Zorbas H
National Institute of Child Health and Human Development, Building 6, Room 416, National Institutes of Health, Bethesda, MD 20892-2753, USA.
Nucleic Acids Res. 1998 May 15;26(10):2255-64. doi: 10.1093/nar/26.10.2255.
Intense interest in the biological roles of DNA methylation, particularly in eukaryotes, has produced at least eight different methods for identifying 5-methylcytosine and related modifications in DNA genomes. However, the utility of each method depends not only on its simplicity but on its specificity, resolution, sensitivity and potential artifacts. Since these parameters affect the interpretation of data, they should be considered in any application. Therefore, we have outlined the principles and applications of each method, quantitatively evaluated their specificity,resolution and sensitivity, identified potential artifacts and suggested solutions, and discussed a paradox in the distribution of m5C in mammalian genomes that illustrates how methodological limitations can affect interpretation of data. Hopefully, the information and analysis provided here will guide new investigators entering this exciting field.
人们对DNA甲基化的生物学作用，尤其是在真核生物中的作用有着浓厚的兴趣，这已经产生了至少八种不同的方法来识别DNA基因组中的5-甲基胞嘧啶及相关修饰。然而，每种方法的实用性不仅取决于其简便性，还取决于其特异性、分辨率、灵敏度以及潜在的假象。由于这些参数会影响数据的解读，因此在任何应用中都应予以考虑。为此，我们概述了每种方法的原理和应用，定量评估了它们的特异性、分辨率和灵敏度，识别了潜在的假象并提出了解决方案，还讨论了哺乳动物基因组中m5C分布的一个悖论，该悖论说明了方法学上的局限性如何影响数据的解读。希望这里提供的信息和分析能为刚进入这个令人兴奋的领域的新研究人员提供指导。
