Agarose Gel Electrophoresis

Daniel Voytas1

1 Iowa State University, Ames, Iowa
Publication Name:  Current Protocols in Neuroscience
Unit Number:  Appendix 1N
DOI:  10.1002/0471142301.nsa01ns11
Online Posting Date:  May, 2001
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Abstract

This appendix presents a protocol for separating and purifying DNA fragments between 0.5 and 25 kb. A support protocol describes the use of midigels and minigels.

     
 
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Table of Contents

  • Basic Protocol 1: Resolution of Large DNA Fragments on Agarose Gels
  • Support Protocol 1: Minigels and Midigels
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Resolution of Large DNA Fragments on Agarose Gels

  Materials
  • Electrophoresis buffer: TAE (unit 4.14) or TBE ( appendix 2A)
  • Ethidium bromide solution ( appendix 2A)
  • Agarose, electrophoresis‐grade
  • 10× loading buffer: 20% (w/v) Ficoll 400/0.1 M disodium EDTA, pH 8.0/1% (w/v) SDS/0.25% (w/v) bromphenol blue
  • DNA molecular weight markers (Fig. )
  • Horizontal gel electrophoresis apparatus
  • Gel casting platform
  • Gel combs (slot formers)
  • DC power supply
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Figures

Videos

Literature Cited

Literature Cited
   Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
   Southern, E. 1979. Gel electrophoresis of restriction fragments. Methods Enzymol. 68:152‐176.
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