Selective RNase H cleavage of target RNAs from a tRNA scaffold.

Luc Ponchon, Geneviève Beauvais, Sylvie Nonin-Lecomte, Frédéric Dardel

Laboratoire de Cristallographie et RMN biologiques, Université Paris Descartes, CNRS, Paris, France. luc.ponchon@parisdescartes.fr

Methods in molecular biology (Clifton, N.J.) 2012

filter terms:

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.

Citation

Luc Ponchon, Geneviève Beauvais, Sylvie Nonin-Lecomte, Frédéric Dardel. Selective RNase H cleavage of target RNAs from a tRNA scaffold. Methods in molecular biology (Clifton, N.J.). 2012;941:9-18