Abstract
A change doesn't occur to base sequence of DNA in the process of the human development. However, one fertilized egg differentiates into cells with sixty trillion of abundant individuality. The reason is DNA is methylated. The DNA methylated correctly is necessary for correct differentiation. Aberrant methylation induce various diseases including cancer, mental disorder and lifestyle disease.Elucidating essential of diseases needs analyzing methylation of DNA.Various DNA methylation analysis methods have been developed, among which bisulfite sequencing (BS-seq) is common.BS-seq has problems such as low sensitivity, which can not analyze cell types with insufficient sample amount, and may miss specific methylation of individual cells because it is an averaged data of cell population.Conventional BS-seq, hydroxymethylation (hmC), which is an intermediate of demethylation, is detected as methylation (mC). Since hmC is not recognized as mC in vivo, conventional BS-seq can not perform accurate DNA methylation analysis.New analytical methods are needed to assess pure DNA methylation even from small numbers of cells, such as single cells. In this study, we aim to develop a novel methylation analysis method that enables accurate methylation to be performed even with a small amount of DNA by repeating amplification and methylation of DNA using PCR and DNA (cytosine-5)-methyltransferase 1 (DNMT1).
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