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Study on Dynamics of Ribozyme Binding of Substrate by Stopped-flow method (CAT#: STEM-AC-0025-WXH)

Introduction

Ribozymes (ribonucleic acid enzymes) are RNA molecules that have the ability to catalyze specific biochemical reactions, including RNA splicing in gene expression, similar to the action of protein enzymes. The most common activities of natural or in vitro evolved ribozymes are the cleavage (or ligation) of RNA and DNA and peptide bond formation.




Principle

Stopped-flow is an experimental technique for studying chemical reactions with a half time of the order of 1 ms.
In its simplest form, a stopped-flow mixes two solutions. Small volumes of solutions are rapidly and continuously driven into a high-efficiency Ball mixer, so mixing is completed in just a few microseconds. This mixing process then initiates an extremely fast reaction.

Applications

Typically used to gain an understanding of reaction mechanisms, including drug-binding processes or following protein structural changes, stopped-flow spectroscopy enables the study of fast reactions in solution over timescales in the range of millisecond to hundreds of seconds.

Procedure

1. In stopped-flow experiments, two, three, or four sample solutions are rapidly mixed and injected into an observation cell.
2. When the flow is stopped, the kinetics are recorded with a detector best suited to the chemical properties of the solutions and the information of interest (e.g. particle size, the environment of the fluorophore, chromophore).

Materials

Stopped-Flows