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Real-Time Detection of Polymerase Activity Using Supercritical Angle Fluorescence


Krieg, Alexander; Ruckstuhl, Thomas; Laib, Stephan; Seeger, Stefan (2004). Real-Time Detection of Polymerase Activity Using Supercritical Angle Fluorescence. Journal of Fluorescence, 14(1):75-78.

Abstract

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

Abstract

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

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Item Type:Journal Article, refereed, original work
Communities & Collections:National licences > 142-005
Dewey Decimal Classification:Unspecified
Scopus Subject Areas:Life Sciences > Biochemistry
Social Sciences & Humanities > Clinical Psychology
Social Sciences & Humanities > Social Sciences (miscellaneous)
Social Sciences & Humanities > Sociology and Political Science
Physical Sciences > Spectroscopy
Life Sciences > Clinical Biochemistry
Social Sciences & Humanities > Law
Language:English
Date:1 January 2004
Deposited On:19 Oct 2018 11:50
Last Modified:15 Apr 2021 14:53
Publisher:Springer
ISSN:1053-0509
OA Status:Green
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1023/b:jofl.0000014663.24696.27

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