Enzymatic probing is a rapid, straightforward method for determining which regions of a folded RNA are structurally constrained. It can be carried out using very small amounts of material, and is especially suitable for short RNAs. Here we report a protocol that we have found to be useful and readily adaptable to the evaluation of RNAs up to 150-200 nucleotides in length. Considerations for optimization are also included. In brief, the method includes folding end-labeled RNA into its native conformation, partial digestion with structure-sensitive nucleases, and identification of the cleavage sites by electrophoretic separation of the cleavage fragments. © 2014 Springer Science+Business Media, LLC.
CITATION STYLE
Biondi, E., & Burke, D. H. (2014). RNA structural analysis by enzymatic digestion. Methods in Molecular Biology, 1086, 41–52. https://doi.org/10.1007/978-1-62703-667-2_3
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