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Updated: May 17, 2026

Analysis of RNA Processing Reactions Using Cell Free Systems: 3' End Cleavage of Pre-mRNA Substrates in vitro
Published on: May 3, 2014
Selective RNase H cleavage of target RNAs from a tRNA scaffold
Luc Ponchon1, Geneviève Beauvais, Sylvie Nonin-Lecomte
1Laboratoire de Cristallographie et RMN biologiques, Université Paris Descartes, CNRS, Paris, France. luc.ponchon@parisdescartes.fr
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.
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