Tethered DNA hairpins facilitate electrochemical detection of DNA ligation

Gerhild Zauner; Yating Wang; Manuel Lavesa-Curto; Andrew MacDonald; Andrew G. Mayes; Richard P. Bowater; Julea N. Butt

doi:10.1039/b413556c

Tethered DNA hairpins facilitate electrochemical detection of DNA ligation

Gerhild Zauner, Yating Wang, Manuel Lavesa-Curto, Andrew MacDonald, Andrew G. Mayes, Richard P. Bowater, Julea N. Butt

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24 Citations (Scopus)

Abstract

A novel electrochemical assay for DNA ligase activity is described. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of ‘nicked’ DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin. The results demonstrate the simplicity of integrating electrochemical detection with hairpin based biosensors and illustrate a new approach to the assay of DNA ligases, of which the NAD+-dependent enzymes represent a potential broad spectrum antibacterial drug target.

Original language	English
Pages (from-to)	345-349
Number of pages	5
Journal	The Analyst
Volume	130
Issue number	3
DOIs	https://doi.org/10.1039/b413556c
Publication status	Published - 2005

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10.1039/b413556c

Cite this

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title = "Tethered DNA hairpins facilitate electrochemical detection of DNA ligation",

abstract = "A novel electrochemical assay for DNA ligase activity is described. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of {\textquoteleft}nicked{\textquoteright} DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin. The results demonstrate the simplicity of integrating electrochemical detection with hairpin based biosensors and illustrate a new approach to the assay of DNA ligases, of which the NAD+-dependent enzymes represent a potential broad spectrum antibacterial drug target.",

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T1 - Tethered DNA hairpins facilitate electrochemical detection of DNA ligation

AU - Zauner, Gerhild

AU - Wang, Yating

AU - Lavesa-Curto, Manuel

AU - MacDonald, Andrew

AU - Mayes, Andrew G.

AU - Bowater, Richard P.

AU - Butt, Julea N.

PY - 2005

Y1 - 2005

N2 - A novel electrochemical assay for DNA ligase activity is described. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of ‘nicked’ DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin. The results demonstrate the simplicity of integrating electrochemical detection with hairpin based biosensors and illustrate a new approach to the assay of DNA ligases, of which the NAD+-dependent enzymes represent a potential broad spectrum antibacterial drug target.

AB - A novel electrochemical assay for DNA ligase activity is described. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of ‘nicked’ DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin. The results demonstrate the simplicity of integrating electrochemical detection with hairpin based biosensors and illustrate a new approach to the assay of DNA ligases, of which the NAD+-dependent enzymes represent a potential broad spectrum antibacterial drug target.

U2 - 10.1039/b413556c

DO - 10.1039/b413556c

M3 - Article

VL - 130

SP - 345

EP - 349

JO - Analyst

JF - Analyst

SN - 0003-2654

IS - 3

ER -