Abstract
The electrophoretic mobility shift assay is a useful tool to identify proteins and nucleic acids interactions. Traditionally, the nucleic acids fragments are end-labeled with 32P. We present here the use of fluorescent methodologies for studies of RNA in place of radioactivity. The method is highly sensitive and quantitative with the use of an infrared fluorescence imaging system. Fluorescently labeled primers can be used to monitor protein–RNA interactions by fluorescent mobility shift assays. The simplicity and validity of this approach may have more advantages than that of previous methods that traditionally used hazardous radioisotopes. This method was successfully tested in the study of the interactions between MS2 Coat Protein and its RNA target.
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Acknowledgements
We thank Dr Maohua Xie (State Key Laboratory of Virology, Department of Biochemistry and Molecular Biology, College of Life Sciences, Wuhan University, China) and Kewei Zheng (Laboratory of Biochemistry and Biophysics, College of Life Sciences, Wuhan University) for their generous donating some of the materials and technical advices. This work was supported by Chinese National Natural Science Foundation of China (30571144).
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Wang, K., Gao, Y., Peng, X. et al. Using FAM labeled DNA oligos to do RNA electrophoretic mobility shift assay. Mol Biol Rep 37, 2871–2875 (2010). https://doi.org/10.1007/s11033-009-9841-7
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DOI: https://doi.org/10.1007/s11033-009-9841-7