The use of RNA electrophoretic mobility shift assays (REMSAs) for analysis of RNA-protein interactions have been limited to lengthy assay time and qualitative assessment. To vastly improve assay efficiency, feasibility and quality of data procured from REMSAs, we combine here some of the best-known labeling and electrophoretic techniques. Nucleic acid fragments are end-labeled with fluorescent tags, as opposed to the radioactive or biotin tags. The fluorescent probes may be detected directly from the electrophoresis gel, eliminating the need for cumbersome membrane transfer and immunoblotting. Modifying the REMSA protocol to include low-molarity, lithium borate conductive media and near-infrared-labeled probes allows for a reduction assay time, quantitative comparison between experimental conditions and crisp band resolution (i.e., optimized results).
Brown, Samantha Z; Agostini, Lebaron C; Thomsett, Henry L; and Brody, Jonathan R, "Ultra-fast conductive media for RNA electrophoretic mobility shift assays." (2020). Department of Surgery Faculty Papers. Paper 181.
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This article has been peer-reviewed. It is the authors final published version in Bio Techniques, Volume 68, Issue 2, February 2020. Pages 101-105.
The published version is available at https://doi.org/10.2144/btn-2019-0111. Copyright Brown et.al.