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Assay of Ribozyme–Substrate Cleavage by Anion-Exchange High-Performance Liquid Chromatography

Authors
Journal
Analytical Biochemistry
0003-2697
Publisher
Elsevier
Publication Date
Volume
232
Issue
2
Identifiers
DOI: 10.1006/abio.1995.0008

Abstract

Abstract An HPLC procedure has been developed that can be used to monitor the rate of phosphodiester cleavage of an oligoribonucleotide substrate by an RNA ribozyme with catalytic activity. The method operates at high substrate concentrations (far beyond K m ), thus allowing the reactions to approach V maxof the reaction. This method is an efficient alternative to radioisotope labeling methods.

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