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
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
.