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Chembiochem. 2003 Jul 7;4(7):589-92.

Real-time detection of nucleotide incorporation during complementary DNA strand synthesis.

Author information

1
Physikalisch-Chemisches Institut, Universität Zürich, Winterthurerstrasse 190, 8057 Zürich, Switzerland. sseeger@pci.unizh.ch

Abstract

Real-time observation of DNA strand synthesis by using a supercritical angle fluorescence detection apparatus for surface-selective fluorescence detection is described. DNA template molecules were immobilized on a glass surface and the synthesis of the complementary strand was observed after addition of enzyme, dTTP, dATP, dGTP, and fluorescently labeled dCTP (d, deoxy; TP, triphosphate; T, A, G, and C, nucleobases). The fluorescence increase during the Klenow-fragment-catalyzed polymerization depends on the number of labeled dCTP nucleotides incorporated. The efficiency of this reaction is of the same order of magnitude as that of a bimolecular hybridization reaction.

PMID:
12851927
DOI:
10.1002/cbic.200200549
[Indexed for MEDLINE]
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