Splint ligation of RNA with T4 DNA ligase

Christopher J. Kershaw, Raymond T. O'Keefe

    Research output: Contribution to journalArticlepeer-review

    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. © 2012 Springer Science+Business Media, LLC.
    Original languageEnglish
    Pages (from-to)257-269
    Number of pages12
    JournalMethods in Molecular Biology
    Volume941
    DOIs
    Publication statusPublished - 2012

    Keywords

    • in vitro transcription
    • RNA ligation
    • RNA modification
    • RNA purification
    • T4 DNA ligase
    • T7 RNA polymerase

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