Recombinant and In Vitro RNA Synthesis pp 9-18

Part of the Methods in Molecular Biology book series (MIMB, volume 941)

| Cite as

Selective RNase H Cleavage of Target RNAs from a tRNA Scaffold

  • Luc Ponchon
  • Geneviève Beauvais
  • Sylvie Nonin-Lecomte
  • Frédéric Dardel
Protocol

Abstract

In vivo overproduction of tRNA chimeras yields an RNA insert within a tRNA scaffold. For some applications, it may be necessary to discard the scaffold. Here we present a protocol for selective cleavage of the RNA of interest from the tRNA scaffold, using RNase H and two DNA oligonucleotides. After cleavage, we show that the RNA of interest can be isolated in a one-step purification. This method has, in particular, applications in structural investigations of RNA.

Key words

Recombinant RNA tRNA scaffold RNase H cleavage RNA purification 

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Copyright information

© Springer Science+Business Media, LLC 2013

Authors and Affiliations

  • Luc Ponchon
    • 1
  • Geneviève Beauvais
    • 2
  • Sylvie Nonin-Lecomte
    • 1
  • Frédéric Dardel
    • 2
  1. 1.Laboratoire de Cristallographie et RMN biologiquesUniversité Paris Descartes, CNRSParisFrance
  2. 2.Laboratoire de Cristallographie et RMN biologiquesUniversité Paris Descartes CNRSParisFrance

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