| 초록 |
A novel electrochemical real-time PCR method has been developed using aptamer-modified primer. For this purpose, Hg2+ ions were employed which bind to thymine-thymine (T-T) mismatched nucleic acids. The 5’ ends of forward (F) and reverse (R) primers were modified with thymine (T)-rich sequences to form ion-induced hairpin structure. In the presence of the target DNA, the aptamer-modified F/R primers can be elongated by the polymerase. After denaturation, the aptamer-modified R/F primers are annealed to extended template and elongated again respectively. As the elongation is completed including aptamer region at the 5’ end, Hg2+ ions are released from the aptamer region of primers. Since the diffusion rate of free ions is higher than that of captured ions, the release of Hg2+ ions can be identified by measuring squarewave voltammetry (SWV). Using this method, we successfully determined target DNA concentration. |
