Splint ligation of RNA, whereby specific RNA molecules are ligated together, can be carried out using T4 DNA ligase and a bridging DNA oligonucleotide complementary to the RNAs. This method takes advantage of the property of T4 DNA ligase to join RNA molecules when they are in an RNA:DNA hybrid. Splint ligation is a useful tool for the introduction of modified nucleotides into RNA molecules, insertion of a radiolabel into a specific position within an RNA and for the assembly of smaller synthetic RNAs into longer RNA molecules. Such modifications enable a wide range of experiments to be carried out with the modified RNA including structural studies, co-immunoprecipitations, and the ability to map sites of RNA:RNA and RNA:protein interactions. © 2012 Springer Science+Business Media, LLC.
CITATION STYLE
Kershaw, C. J., & O’Keefe, R. T. (2012). Splint ligation of RNA with T4 DNA ligase. Methods in Molecular Biology, 941, 257–269. https://doi.org/10.1007/978-1-62703-113-4_19
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