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

doi:10.2116/analsci.32.505

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

Department of Applied Chemistry

Research output: Contribution to journal › Article › peer-review

7 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 language	English
Pages (from-to)	505-510
Number of pages	6
Journal	analytical sciences
Volume	32
Issue number	5
DOIs	https://doi.org/10.2116/analsci.32.505
Publication status	Published - 2016

Keywords

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

ASJC Scopus subject areas

Analytical Chemistry

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10.2116/analsci.32.505

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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 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.",

keywords = "Ion-selective electrode, Isothermal DNA amplification, PG-RCA, Potentiometry, Real-time monitoring",

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

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doi = "10.2116/analsci.32.505",

language = "English",

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AU - Seichi, Ayaka

AU - Kozuka, Nanami

AU - Kashima, Yuko

AU - Tabata, Miyuki

AU - Goda, Tatsuro

AU - Matsumoto, Akira

AU - Iwasawa, Naoko

AU - Citterio, Daniel

AU - Miyahara, Yuji

AU - Suzuki, Koji

N1 - Publisher Copyright: © The Japan Society for Analytical Chemistry.

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

KW - Ion-selective electrode

KW - Isothermal DNA amplification

KW - PG-RCA

KW - Potentiometry

KW - Real-time monitoring

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JO - analytical sciences

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ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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