Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4031

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Restriction Endonuclease Digestion of DNA in Agarose Plugs

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

Genomic DNA isolated from mammalian, yeast, or bacterial cells can be digested with restriction endonucleases by incubating agarose plugs containing the DNA in the presence of the desired enzyme. After digestion, the DNA can be fractionated by pulsed-field gel electrophoresis and either isolated from the gel or analyzed by Southern Hybridization.


MATERIALS

Genomic DNA embedded in low-melting-temperature agarose plugs

Please see Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells and Tissuesfor the preparation of low-melting-temperature agarose plugs containing mammalian cell DNA . . . [Full Text of this Article]


METHOD


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