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Application of Microchip Electrophoresis in the Analysis of RNA Aptamer-Protein Interactions
Authors:Fumiko Nishikawa  Hidetoshi Arakawa  Satoshi Nishikawa
Institution:1. Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST) , Tsukuba , Ibaraki , Japan;2. School of Pharmaceutical Sciences, Showa University , Shinagawa, Tokyo , Japan;3. Age Dimension Research Center , AIST , Tsukuba , Ibaraki , Japan
Abstract:

DNA and RNA can be separated by microchip electrophoresis (ME) and detected using an intercalating fluorescent dye. The advantages of this method are short sensing times (< 3 min), avoidance of a radioisotope labeling detection system, relatively low costs, and reduced labor intensity. In the present study, RNA aptamer-protein or -peptide interactions were analyzed using ME and the regression of free aptamers corresponding to unbound RNA was detected as the target protein or peptide increased in a dose-dependent manner. Our results demonstrate the applicability of this method to simple, rapid ligand screening in the interactions between oligonucleotides and their targets.
Keywords:In vitro selection  Microchip electrophoresis  RNA aptamer  SYBR gold
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