An electrochemical gene detection assay utilising T7 exonuclease activity on complementary probe-target oligonucleotide sequences

S C Hillier, Stephen E Flower, Christopher G Frost, A Toby A Jenkins, R Keay, H Braven, J Clarkson

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

This communication describes the synthesis of an electrochemically active oligonucleotide probe and its application in sensing complementary oligonucleotides sequences using a T7 exonuclease enzyme. Target oligonucleotides are detected by hybridisation with a ferrocene labelled probe oligonucleotide followed by addition of T7 exonuclease. The T7 enzyme is a double strand specific exonuclease that removes the terminal 5' nucleotide of the probe sequence. The 5' nucleotide is attached to a ferrocene label, which is subsequently detected at an electrode using differential pulse voltammetry. Time and temperature resolved measurements were performed and an associated study using dual labelled fluorophore-quencher labelled probes was performed to confirm the validity of the electrochemical assay.
Original languageEnglish
Pages (from-to)1227-1232
Number of pages6
JournalElectrochemistry Communications
Volume6
Issue number12
Early online date17 Oct 2004
DOIs
Publication statusPublished - 1 Dec 2004

Keywords

  • DNA
  • electrochemistry
  • T7 exonuclease

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