Development ofa novel method for analyzing various DNA methylation

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Microbes harbor various methylated DNA species, such as 5-methylcytosine, N6-methyladenine, and N4-methylcytosine, which are produced by DNA restriction-modification systems. Aiming at the development of a novel method for widely analyzing DNA methylation sites, I designed immunological methods to analyze N6-methyladenine sites arising from type I restriction-modification systems (immunoprecipitation/PCR, methylated plasmid library/HPLC, methylated plasmid library/western blot) and evaluated them. Detailed analysis suggested that the method “methylated plasmid library/western blot” was effective for analyzing type I methylation sites.