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Real-Time Detection of Polymerase Activity Using Supercritical Angle Fluorescence

Krieg, Alexander ; Ruckstuhl, Thomas ; Laib, Stephan ; Seeger, Stefan

In: Journal of Fluorescence, 2004, vol. 14, no. 1, p. 75-78

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    Titre
    • Real-Time Detection of Polymerase Activity Using Supercritical Angle Fluorescence
    Auteur
    • Krieg, Alexander. Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstr. 190, 8057, Zürich, Switzerland
    • Ruckstuhl, Thomas. Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstr. 190, 8057, Zürich, Switzerland
    • Laib, Stephan. Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstr. 190, 8057, Zürich, Switzerland
    • Seeger, Stefan. Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstr. 190, 8057, Zürich, Switzerland
    Type de document
    • Postprint
    Identifiant OAI-PMH
    • oai:doc.rero.ch:319981
    Summary
    We investigated the incorporation efficiencies of different fluorescently labelled dNTPs with polymerases by complementary strand synthesis. For this reason single stranded DNA was immobilized on a coverslip and the increase of fluorescence due to the synthesis of the corresponding strand with tagged dNTPs was detected with a supercritical angle fluorescence biosensor in real-time. By comparison of the observed signal intensities it was possible to conclude that the system Cy5-dCTP—Klenow (exonuclease free) fragment gives the best incorporation yield of the investigated enzymes and dNTPs