Journal of Fluorescence

, Volume 15, Issue 3, pp 207–214

A New Fluorescence Resonance Energy Transfer Pair and Its Application to Oligonucleotide Labeling and Fluorescence Resonance Energy Transfer Hybridization Studies

  • Michaela Gruber
  • Bianca Wetzl
  • Bernhard Oswald
  • Joerg Enderlein
  • Otto S. Wolfbeis
Article

DOI: 10.1007/s10895-005-2619-y

Cite this article as:
Gruber, M., Wetzl, B., Oswald, B. et al. J Fluoresc (2005) 15: 207. doi:10.1007/s10895-005-2619-y

Abstract

We describe two new fluorescence resonance energy transfer (FRET) compatible labels, their covalent linkage to oligonucleotides, and their use as donor and acceptor, respectively, in FRET hybridization studies. The dyes belong to the cyanine dyes, and water solubility is imparted by a phosphonate which represents a new solubilizing group in DNA labels. They were linked to amino-modified synthetic oligonucleotides via oxysuccinimide (OSI) esters. The studies performed include binding assays, determinations of molecular distances, homogeneous competitive assays, and limits of detection, which are in the order of 5 pmol/L for a 15-mer.

Key Words

Fluorescence hybridization assay fluorescence resonance energy transfer long-wavelength fluorescent labels 

Abbreviations:

BCBS

bicarbonate buffer solution, 50 mM, pH 9

DCC

dicyclohexylcarbodiimide

DCM

dichloromethane

FRET

fluorescence resonance energy transfer

NHS

N-hydroxysuccinimide

OSI

oxysuccinimidyl

PBS

phosphate-buffered saline, 22 mM, pH 7.2

trisacetate buffer

50 mM, pH 7.2 with 25 mM NaCl

Copyright information

© Springer Science + Business Media, Inc. 2005

Authors and Affiliations

  • Michaela Gruber
    • 1
    • 2
  • Bianca Wetzl
    • 1
  • Bernhard Oswald
    • 2
  • Joerg Enderlein
    • 3
  • Otto S. Wolfbeis
    • 1
  1. 1.Institute of Analytical Chemistry, Chemo- and BiosensorsUniversity of RegensburgRegensburgGermany
  2. 2.Chromeon GmbHRegensburgGermany
  3. 3.Forschungszentrum Jülich, Institute of Biological Information Processing IJülichGermany

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