title: Real-time Monitoring and Detection of Primer Generation-Rolling Circle Amplification of DNA Using an Ethidium Ion-selective Electrode.
abstract: An electrochemical detection system for an isothermal DNA amplification method using an ion-selective electrode (ISE) was developed as a low-cost, simple and real-time monitoring system. The system is based on potentiometry using an ethidium ion (Et(+)) selective electrode that relies on monitoring DNA amplification by measuring potential changes in the reaction solution containing ethidium bromide (EtBr) as an intercalator to DNA. With progressing primer generation-rolling circle amplification (PG-RCA) under isothermal condition at 37°C, EtBr is bound to the newly formed DNA, resulting in a lowered free EtBr concentration in the sample solution. In this case, the Et(+) ISE potential allows real-time monitoring of the PG-RCA reaction in the range of 10 nM - 1 μM initial target DNA.
authors.affiliation: Department of Applied Chemistry, Graduate School of Science and Engineering, Keio University.
authors.forename: Ayaka
authors.initials: A
authors.lastname: Seichi
authors.forename: Nanami
authors.initials: N
authors.lastname: Kozuka
authors.forename: Yuko
authors.initials: Y
authors.lastname: Kashima
authors.forename: Miyuki
authors.initials: M
authors.lastname: Tabata
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Goda
authors.forename: Akira
authors.initials: A
authors.lastname: Matsumoto
authors.forename: Naoko
authors.initials: N
authors.lastname: Iwasawa
authors.forename: Daniel
authors.initials: D
authors.lastname: Citterio
authors.forename: Yuji
authors.initials: Y
authors.lastname: Miyahara
authors.forename: Koji
authors.initials: K
authors.lastname: Suzuki
mesh_terms: D055664:Electrochemical Techniques / Q000295:instrumentation*; D004996:Ethidium / Q000737:chemistry*; D017736:Ion-Selective Electrodes; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000295:instrumentation*
chemical_list: D004996:Ethidium