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RNA Modification

RNA can be modified with enzymes and reagents that act selectively depending on the existence of particular structures, and hence are useful tools for characterizing RNA species. The selective properties of RNA modifying enzymes enable researchers to distinguish different RNA species. Some unique end modifications can be used to selectively degrade or isolate particular RNA species when treated sequentially with the appropriate series of enzymes. For instance, 5´-capped RNAs are not substrates for polynucleotide kinases, or RNA ligases, but are substrates for decapping enzymes. Similarly, ligation and poly-adenylation of RNA 3´ end requires a free 3´.

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Protocols for RNA Modification
    Publications related to RNA Modification
  1. Eyler, D.E., Franco, M.K., Batool, Z., Wu, M.Z., Dubuke, M.L., Dobosz-Bartoszek, M., Jones, J.D., Polikanov, Y.S., Roy, B., Koutmou, K.S 2019. Pseudouridinylatio of mRNA coding sequences alters translation Proc Natl Acad Sci U S A. 116(46), PubMedID: 31672910, DOI: 10.1073/pnas.1821754116
  2. Wulf, Madalee; Buswell, John; Chan, Siuhong; Dai, Nan; Marks, Katherine; Tzertzinis, George; Whipple, Joe; Correa, Ivan; Schildkraut, Ira; 2019. The yeast scavenger decapping enzyme DcpS and its application for in vitro RNA recapping Sci Rep. 9 (1), PubMedID: 31197197, DOI: 10.1038/s41598-019-45083-5
  3. Potapov, V., Fu, X., Dai, N., Correa, I.R., Jr., Tanner, N.A., Ong, J.L 2018. Base modificatons affecting RNA polymerase and reverse transcriptase fidelity Nucleic Acids Res. 46(11): 5753-5763, PubMedID: 29750267, DOI:
RNA Ligase Selection Chart
NEB offers a variety of ligases for RNA research with unique specificities.
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Videos

  1. invitro_gaussia_video_tn

    In Vitro Transcription and Capping of Gaussia Luciferase mRNA Followed by HeLa Cell Transfection

    This method describes high yield in vitro synthesis of both capped and uncapped mRNA from a linearized plasmid containing the Gaussia luciferase (GLuc) gene. 

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