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Proceedings Paper

Excited-state quenching of dye-linked oligonucleotides
Author(s): David P. Millar; Remo A. Hochstrasser; Christopher R. Guest; Shiow Meei Chen
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Paper Abstract

Fluorescence energy transfer (FET) can be used to obtain structural and dynamic information on duplex and branched DNA molecules suitably labeled with donor and acceptor dyes. However, dye-DNA interaction cause excited-state quenching of the dyes and complicate the analysis of FET data. We have investigated the excited-state quenching of two common FET dyes, fluorescein and tetramethyl rhodamine, covalently linked to synthetic oligonucleotides. The rate of quenching is shown to depend on the base sequence and association state of the oligonucleotide, and also the length of the linker chain. Furthermore, the dyes can adopt more than one configuration with respect to the oligonucleotide strand and the degree of quenching is different in each configuration. The implications of these findings for FET measurements in nucleic acids are discussed.

Paper Details

Date Published: 1 April 1992
PDF: 7 pages
Proc. SPIE 1640, Time-Resolved Laser Spectroscopy in Biochemistry III, (1 April 1992); doi: 10.1117/12.58254
Show Author Affiliations
David P. Millar, Scripps Research Institute (United States)
Remo A. Hochstrasser, Scripps Research Institute (United States)
Christopher R. Guest, Scripps Research Institute (United States)
Shiow Meei Chen, Scripps Research Institute (United States)


Published in SPIE Proceedings Vol. 1640:
Time-Resolved Laser Spectroscopy in Biochemistry III
Joseph R. Lakowicz, Editor(s)

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