Organic Chemistry I - Bioorganic Chemistry, Institute of Chemistry, University of Osnabrück, Barbarastrasse 7, D-49069 Osnabrück.
Chem Biodivers. 2012 Feb;9(2):272-81. doi: 10.1002/cbdv.201100298.
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.
描述了一种新的技术，该技术包括在脂质双层-H2O 相界面层处形成碱基特异性 DNA 双链。两种不同的带有双尾脂质的寡核苷酸探针都在 5'端被整合到稳定的人工脂质双层中，该双层将具有灌注能力的光学透明微流控样品载体的两个隔室（顺式/反式通道）隔开。顺式和反式通道都充满盐水缓冲液。将与寡核苷酸探针之一互补的 Cy5 标记的靶 DNA 序列注入顺式通道，然后彻底灌注，通过单分子荧光光谱和显微镜检测到标记的互补寡核苷酸在膜上的固定化。对于荧光但不互补的 DNA 序列，在膜上未检测到固定化的荧光寡核苷酸双链。这清楚地证实了在膜界面处的特异性双链形成。
