Splint ligation of RNA with T4 DNA ligase.
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.
The University of Manchester, Manchester, UK.
Christopher J
CJ
Kershaw
Raymond T
RT
O'Keefe
D017122:Bacteriophage T4 / Q000201:enzymology; D011088:DNA Ligases / Q000378:metabolism*; D006151:Guanosine / Q000378:metabolism; D006157:Guanosine Monophosphate / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D012313:RNA / Q000096:biosynthesis / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000378:metabolism; D014158:Transcription, Genetic
D012333:RNA, Messenger; D006151:Guanosine; D012313:RNA; D006157:Guanosine Monophosphate; D006160:Guanosine Triphosphate; D011088:DNA Ligases