Analytical Sciences

Sequence-selective Modification of DNA Cytosine by Using Junction-forming DNA Probes and Its Application to the Detection of Single Cytosine Methylation

Kenta TAKANASHI and Teru KATO

Graduate School of Bionics, Computer and Media Sciences, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan

Methylated DNA plays an important role in epigenetic gene regulation, and could therefore serve as a potentially promising biomarker for the diagnosis of cancer and other diseases. Therefore, the development of a simple method for analyzing cytosine methylation in a target gene is required. Here, we report on a new method for the sequence-selective chemical modification of a single cytosine in single-stranded DNA (ssDNA). This method is based on the formation of a DNA three-way junction of the ssDNA and two ssDNA probes, and was successfully applied for a simple polymerase chain reaction (PCR)-based assay for ‘pinpoint’ methylation analysis.

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