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A New Fluorescence Resonance Energy Transfer Pair and Its Application to Oligonucleotide Labeling and Fluorescence Resonance Energy Transfer Hybridization Studies
Authors:Michaela?Gruber,Bianca?Wetzl,Bernhard?Oswald,Joerg?Enderlein,Otto?S.?Wolfbeis  author-information"  >  author-information__contact u-icon-before"  >  mailto:otto.wolfbeis@chemie.uni-regensburg.de"   title="  otto.wolfbeis@chemie.uni-regensburg.de"   itemprop="  email"   data-track="  click"   data-track-action="  Email author"   data-track-label="  "  >Email author
Affiliation:(1) Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040 Regensburg, Germany;(2) Chromeon GmbH, Josef-Engert-Strasse 9, 93053 Regensburg, Germany;(3) Forschungszentrum Jülich, Institute of Biological Information Processing I, 52425 Jülich, Germany
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.
Keywords:Fluorescence hybridization assay  fluorescence resonance energy transfer  long-wavelength fluorescent labels
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