Abstract
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.
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Acknowledgments
This work was supported by the Biotechnology and Biological Sciences Research Council (BBSRC) and The Wellcome Trust.
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Kershaw, C.J., O’Keefe, R.T. (2013). Splint Ligation of RNA with T4 DNA Ligase. In: Conn, G. (eds) Recombinant and In Vitro RNA Synthesis. Methods in Molecular Biology, vol 941. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-113-4_19
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DOI: https://doi.org/10.1007/978-1-62703-113-4_19
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Publisher Name: Humana Press, Totowa, NJ
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