earticle

영어

DNA methylation at CpG dinucleotides residues is known as central mechanism to control gene expression. As aberrant DNA methylation is known to be genetic cause of many kinds of diseases, it is a strong candidate for biomarker for diagnosis.Although various technologies have been used to analyze DNA methylation, those methods are insufficient as a routine diagnosis tool because of potential error by incomplete conversion of DNA. Methylation sensitive (MS-) multiplex ligation-dependent probe amplification (MLPA) has a potential as a diagnosis tool, because it is a conversion-free method. Additionally, MS-MLPA can analyze multiple targets, which is essential for diagnosis of most diseases. But this method has limitations: designing a custom MLPA probe is more difficult due to the stuffer sequence, and only the limited target site can be analyzed due to the sequence specific restriction enzyme. In this study, we developed a novel DNA methylation analysis method using MS-MLPA, in which stuffer-free MLPA probes and McrBC enzyme are used for precise multiplex quantitative analysis. We tested our new method to detect the methylation statues of 4 tumor suppress genes.