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Application of a Highly Robust and Efficient Fluorescence‐Resonance‐Energy‐Transfer (FRET) System in DNA
Authors:Lilia Clima  Caroline Hirtz‐Haag  Andrea Kienzler  Willi Bannwarth
Affiliation:1. Institut für Organische Chemie und Biochemie, Albert‐Ludwigs Universit?t Freiburg, Albertstr. 21, D‐79104 Freiburg (phone: +49?761?2036073;2. fax: +49?761?2038705)
Abstract:We report a feasibility study for the application of our newly developed highly efficient and robust fluorescence‐resonance‐energy‐transfer (FRET) system to DNA. A 2′‐oligodeoxynucleotide, 12 , equipped with a quinolinone derivative as donor and a (bathophenanthroline)ruthenium(II) complex as acceptor and having a single uridine as potential cleavage site under basic conditions revealed an intensive FRET, which almost vanished after cleavage of the oligonucleotide under basic conditions (Fig. 7). Furthermore, in the arrangement of a molecular beacon (MB) DNA (see 13 ), a significant decrease of the FRET was observed after hybridization to a target sequence (Fig. 9). Due to the long decay times of the fluorescence of the Ru‐complex, the system allows for highly sensitive time‐gated measurements.
Keywords:DNA  Fluorescence resonance energy transfer (FRET)  Molecular beacon  Nucleic acids  Time‐resolved FRET
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