Abstract
Small RNAs, defined as noncoding 20–30-nt-long RNAs, are instrumental regulators of cellular processes in most eukaryotes. In this chapter we describe three methods for extracting small RNA from cells: a general method, one plant specific and a third particular to conifers. Further, protocols are given for the analysis and quantification of small RNAs using polyacrylamide gel-based approaches. A native streptavidin gel-shift assay, useful for measuring the relative amounts of multiple small RNAs simultaneously, is presented. To further characterize small RNAs biochemically, a sodium periodate assay probing for 2′, 3′ hydroxyl groups on the 3′ terminus of small RNAs is outlined.
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Acknowledgments
Marie Curie International Incoming Fellow from the European Commission, Community Research and Development Information Service, Seventh Framework Programme (to H.A.E.). Operating grants from the Natural Sciences and Engineering Research Council of Canada and the Michael Smith Foundation (to P.J.U.).
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Ebhardt, H.A., Ovando, M.O., Unrau, P.J. (2012). Isolation and Biochemical Analysis of Plant Small RNAs. In: Watson, J., Wang, MB. (eds) Antiviral Resistance in Plants. Methods in Molecular Biology, vol 894. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-882-5_15
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DOI: https://doi.org/10.1007/978-1-61779-882-5_15
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