High-performance liquid chromatography purification of homogenous-length RNA produced by trans cleavage with a hammerhead ribozyme

Citation
Tp. Shields et al., High-performance liquid chromatography purification of homogenous-length RNA produced by trans cleavage with a hammerhead ribozyme, RNA, 5(9), 1999, pp. 1259-1267
Citations number
27
Categorie Soggetti
Biochemistry & Biophysics
Journal title
RNA-A PUBLICATION OF THE RNA SOCIETY
ISSN journal
13558382 → ACNP
Volume
5
Issue
9
Year of publication
1999
Pages
1259 - 1267
Database
ISI
SICI code
1355-8382(199909)5:9<1259:HLCPOH>2.0.ZU;2-I
Abstract
An improved method is presented for the preparation of milligram quantities of homogenous-length RNAs suitable for nuclear magnetic resonance or X-ray crystallographic structural studies, Heterogeneous-length RNA transcripts are processed with a hammerhead ribozyme to yield homogenous-length product s that are then readily purified by anion exchange high-performance liquid chromatography. This procedure eliminates the need for denaturing polyacryl amide gel electrophoresis, which is the most laborious step in the standard procedure for large-scale production of RNA by in vitro transcription, The hammerhead processing of the heterogeneous-length RNA transcripts also sub stantially improves the overall yield and purity of the desired RNA product .