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08. Dezember 2014 : Specific DNA duplex formation at an artificial lipidbilayer: fluorescence microscopy after Sybr Green Istaining

Emma Werz1,2 and Helmut Rosemeyer*1 Beilstein J. Org. Chem., 2014, 10, 2307-2321 doi:10.3762/bjoc.10.240

The article describes the immobilization of different probe oligonucleotides (4,7,10) carrying each aracemic mixture of 2,3-bis(hexadecyloxy)propan-1-ol(1a)atthe5’-terminus on a stable artificial lipid bilayer composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The bilayer separates two compartments(cis/transchannel) of an optical transparent microfluidic sample carrier with perfusion capabilities. Injection of unlabeled target DNA sequences (6, 8, or 9), differing in sequence and length, leads in the case of complementarity to the formationof stable DNA duplexes at the bilayer surface. This could be verified by Sybr Green I double strand staining, followed by incubation periods and thorough perfusions, and was visualized by single molecule fluorescence spectroscopy and microscopy. The different bilayer-immobilized complexes consisting of various DNA duplexes and the fluorescent dye were studied with respect to the kinetics of their formation as well as to their stability against perfusion.