Specific DNA duplex formation at an artificial lipid bilayer: towards a new DNA biosensor technology |
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Authors: | Werz Emma Korneev Sergei Montilla-Martinez Malayko Wagner Richard Hemmler Roland Walter Claudius Eisfeld Jörg Gall Karsten Rosemeyer Helmut |
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Institution: | Organic Chemistry I - Bioorganic Chemistry, Institute of Chemistry, University of Osnabrück, Barbarastrasse 7, D-49069 Osnabrück. |
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Abstract: | A novel technique is described which comprises a base-specific DNA duplex formation at a lipid bilayer-H(2) O-phase boundary layer. Two different probes of oligonucleotides both carrying a double-tailed lipid at the 5'-terminus were incorporated into stable artificial lipid bilayers separating two compartments (cis/trans-channel) of an optically transparent microfluidic sample carrier with perfusion capabilities. Both the cis- and trans-channels are filled with saline buffer. Injection of a cyanine-5-labeled target DNA sequence, which is complementary to only one of the oligonucleotide probes, into the cis-channel, followed by a thorough perfusion, leads to an immobilization of the labeled complementary oligonucleotide on the membrane as detected by single-molecule fluorescence spectroscopy and microscopy. In the case of fluorescent but non-complementary DNA sequences, no immobilized fluorescent oligonucleotide duplex could be detected on the membrane. This clearly verifies a specific duplex formation at the membrane interface. |
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