Abstract
In RNA nanotechnology, construction of nanoparticles involves conjugation of functionalities, cross-linking of modules, labeling of RNA subunits, and chemical modification of nucleotides. Efficiency and sensitivity are important for the RNA labeling, which also can be used as probes in microarrays, Northern blotting, and gel-shift assays. Here, we describe a method for fluorescence labeling of short RNA at the 3′-end by oxidation. The 3′-terminus of in vitro-transcribed short RNA is oxidized by sodium periodate, and fluorescein-5-thiosemicarbazide is added after removal of excess oxidant. Purified short RNA with fluorescence is then applied for detection of RNA–protein interaction by gel-shift assay.
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Acknowledgement
This work was supported by grants from National Natural Science Foundation of China (31270842) and National Basic Research Program of China (2012CB114101) to YZX.
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Qiu, C., Liu, WY., Xu, YZ. (2015). Fluorescence Labeling of Short RNA by Oxidation at the 3′-End. In: Guo, P., Haque, F. (eds) RNA Nanotechnology and Therapeutics. Methods in Molecular Biology, vol 1297. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2562-9_8
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DOI: https://doi.org/10.1007/978-1-4939-2562-9_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2561-2
Online ISBN: 978-1-4939-2562-9
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