Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4033

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Pulsed-field Gel Electrophoresis via Contour-clamped Homogeneous Electric Field Gels

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

In CHEF gels, the electric field is generated from multiple electrodes, arranged in a square of hexagonal contour around the horizontal gel and clamped to predetermined potentials. Using a combination of low field strengths, low concentrations of aragose, long switching intervals, and extended periods of electrophoresis, DNAs up to 5000 kb can be resolved. This protocol, supplied by Elsy Jones (University of Texas Southwestern Medical Center, Dallas), describes the resolution of genomic DNA by TAFE, followed by blotting and hybridization.


MATERIALS

recipe 0.5x TBE gel electrophoresis buffer

DNA size standards

Please see Step 3.

recipe DNA staining solution

For a discussion of . . . [Full Text of this Article]


METHOD


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