Real-time monitoring and detection of primer generation-rolling circle amplification of DNA using an ethidium ion-selective electrode

Ayaka Seichi, Nanami Kozuka, Yuko Kashima, Miyuki Tabata, Tatsuro Goda, Akira Matsumoto, Naoko Iwasawa, Daniel Citterio, Yuji Miyahara, Koji Suzuki

Research output: Contribution to journalArticle

5 Citations (Scopus)

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 generationrolling 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.

Original languageEnglish
Pages (from-to)505-510
Number of pages6
Journalanalytical sciences
Volume32
Issue number5
DOIs
Publication statusPublished - 2016

Keywords

  • Ion-selective electrode
  • Isothermal DNA amplification
  • PG-RCA
  • Potentiometry
  • Real-time monitoring

ASJC Scopus subject areas

  • Analytical Chemistry

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  • Cite this

    Seichi, A., Kozuka, N., Kashima, Y., Tabata, M., Goda, T., Matsumoto, A., Iwasawa, N., Citterio, D., Miyahara, Y., & Suzuki, K. (2016). Real-time monitoring and detection of primer generation-rolling circle amplification of DNA using an ethidium ion-selective electrode. analytical sciences, 32(5), 505-510. https://doi.org/10.2116/analsci.32.505