Study of enzymatic cleavage of covalently closed circular DNA by Quench-Flow method (CAT#: STEM-AC-0051-WXH)

Introduction

cccDNA (covalently closed circular DNA) is a special DNA structure that arises during the propagation of some viruses in the cell nucleus and may remain permanently there. It is a double-stranded DNA that originates in a linear form that is ligated by means of DNA ligase to a covalently closed ring.

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Principle Applications Procedure Materials Notes Related Products

Principle

The principle is to mix the two solutions and then to observe a change in spectroscopic properties of the mixture at different place along the reaction tube during the flow. In the continuous mode of the quenched-flow method, the observation chamber is replaced by a second mixer in which the quenching agent arrives.

Applications

Used to determine fast reaction rates or single turnover rates of enzymatic reactions and to isolate reaction intermediates.

Procedure

Small volumes of solutions are driven through a high efficiency mixer and flow into a delay (or ageing) loop. After a set time, the reaction is stopped (or quenched) by the addition of a chemical quench solution.

Materials

Quench flow

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