Hengesbach Martin, Kobitski Andrei, Voigts-Hoffmann Felix, Frauer Carina, Nienhaus G Ulrich, Helm Mark
Institute of Pharmacy and Molecular Biotechnology, University of Heidelberg, Heidelberg, Germany.
Curr Protoc Nucleic Acid Chem. 2008 Sep;Chapter 11:Unit 11.12. doi: 10.1002/0471142700.nc1112s34.
Investigation of single RNA molecules using fluorescence resonance energy transfer (FRET) is a powerful approach to investigate dynamic and thermodynamic aspects of the folding process of a given RNA. Its application requires interdisciplinary work from the fields of chemistry, biochemistry, and physics. The present work gives detailed instructions on the synthesis of RNA molecules labeled with two fluorescent dyes interacting by FRET, as well as on their investigation by single-molecule fluorescence spectroscopy.
利用荧光共振能量转移(FRET)对单个RNA分子进行研究,是探究特定RNA折叠过程的动力学和热力学方面的有力方法。其应用需要化学、生物化学和物理领域的跨学科合作。本工作详细介绍了用两种通过FRET相互作用的荧光染料标记的RNA分子的合成方法,以及通过单分子荧光光谱对其进行研究的方法。