Analytical Sciences
Online ISSN : 1348-2246
Print ISSN : 0910-6340
Original Papers
Real-time Monitoring and Detection of Primer Generation-Rolling Circle Amplification of DNA Using an Ethidium Ion-selective Electrode
Ayaka SEICHINanami KOZUKAYuko KASHIMAMiyuki TABATATatsuro GODAAkira MATSUMOTONaoko IWASAWADaniel CITTERIOYuji MIYAHARAKoji SUZUKI
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2016 Volume 32 Issue 5 Pages 505-510

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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.

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© 2016 by The Japan Society for Analytical Chemistry
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