Use of high-performance liquid chromatographic fractionation of large RNA molecules in the assay of group I intron ribozyme activity

Ion-pair reversed-phase high-performance liquid chromatography (HPLC), whic h has previously been used to fractionate double-stranded DNA molecules, ca n be applied to single-stranded RNA molecules in the size range of 200-1000 nucleotides. This procedure permits RNA molecules to be separated and reco vered rapidly in liquid medium, thereby facilitating recovery. We have used this system to separate an in vitro transcription product containing a gro up I intron ribozyme from the intermediates and products of the splicing re action, permitting rapid assay of ribozyme activity without the use of radi oactivity. (C) 2000 Elsevier Science BN. All rights reserved.