Selective RNase H cleavage of target RNAs from a tRNA scaffold.
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.
Laboratoire de Cristallographie et RMN biologiques, Université Paris Descartes, CNRS, Paris, France. luc.ponchon@parisdescartes.fr
Luc
L
Ponchon
Geneviève
G
Beauvais
Sylvie
S
Nonin-Lecomte
Frédéric
F
Dardel
D004591:Electrophoresis, Polyacrylamide Gel; D059367:RNA Cleavage; D012336:RNA, Ribosomal, 16S / Q000302:isolation & purification / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D016914:Ribonuclease H / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013194:Staining and Labeling
D012336:RNA, Ribosomal, 16S; D012343:RNA, Transfer; D016914:Ribonuclease H