원문정보
초록
영어
The strategy for electrochemical detection of DNA mutation was developed with SURVEYOR nuclease and synthesized signal enhancing material. 50 base-long capture probe having thiol group and biotin at 3’ and 5’-end respectively was immobilized on the gold electrode surface and mercaptohexanol was applied subsequently as a blocking reagent. Perfect matched target and single-base mismatched target was hybridized on the each electrode. SURVEYOR nuclease was added and it cleaved the single-base mismatched dsDNA. The cleaved fraction including biotin at 5’-end was removed during washing step. Glucose oxidase (GOX, signal generating enzyme)-avidin conjugate was reacted biotin at 5’-end of capture probe, while it could not react with single-base mismatched duplex. Additionally, biotin-modified G4 PAMAM (Polyamidoamime) dendrimer was added to increase density of the GOX on the electrode surface. Cyclic voltammetry was performed with electrolyte containing 0.1 mM ferrocenemethanol and 10 mM glucose. The higher portion of single-base mismatched target in whole sample, the lower peak current was observed. With this simple and fast electrochemical system, the possibility of mutation in unknown sample can be detected clearly.