Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3913

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Purification of Closed Circular DNA by Equilibrium Centrifugation in CsCl-Ethidium Bromide Gradients: Discontinuous Gradients

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 15% of the full text of this article appears below.


INTRODUCTION

A solution containing plasmid DNA, saturating amounts of ethidium bromide, and CsCl (44% w/v) is layered between two solutions of lesser (35% w/v CsCl) and greater density (59% w/v CsCl). During centrifugation to equilibrium, the closed circular plasmid DNA and linear DNAs form bands at different densities.


MATERIALS

Crude plasmid preparation

Use material from either Step 16 of Preparation of Plasmid DNA by Alkaline Lysis with SDS: Maxipreparation, Step 18 of Preparation of . . . [Full Text of this Article]


METHOD


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