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

被引：43

作者：

Shields, TP ^{[1
]}

Mollova, E ^{[1
]}

Marie, LS ^{[1
]}

Hansen, MR ^{[1
]}

Pardi, A ^{[1
]}

机构：

[1] Univ Colorado, Dept Chem & Biochem, Boulder, CO 80309 USA

来源：

RNA | 1999年 / 5卷 / 09期

关键词：

hammerhead ribozyme; HPLC; in vitro transcription; RNA synthesis;

D O I：

10.1017/S1355838299990945

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 products that are then readily purified by anion exchange high-performance liquid chromatography. This procedure eliminates the need for denaturing polyacrylamide 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 substantially improves the overall yield and purity of the desired RNA product.

引用

页码：1259 / 1267

页数：9

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