Preparation of Genomic DNA from Mammalian Tissue

William.M. Strauss1

1 Whitehead Institute, Cambridge, Massachusetts
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 10.2
DOI:  10.1002/0471142735.im1002s08
Online Posting Date:  May, 2001
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Abstract

There are a number of different procedures for the preparation of genomic DNA. They all start with some form of cell lysis, followed by deproteinization and recovery of DNA. The main differences between various approaches lie in the extent of deproteinization and in molecular weight of the DNA produced. The isolation procedure described here is relatively brief and relies on the powerful proteolytic activity of proteinase K combined with the denaturing ability of the ionic detergent SDS.

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
  • Literature Cited
     
 
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Materials

Basic Protocol 1:

  Materials
  • Liquid nitrogen
  • recipeDigestion buffer
  • Phosphate‐buffered saline (PBS; appendix 2A), ice‐cold
  • 25:24:1 phenol/chloroform/isoamyl alcohol (unit 10.1)
  • 7.5 M ammonium acetate
  • 100% and 70% ethanol
  • TE buffer, pH 8 ( appendix 2A)
  • Sorvall H1000B rotor (or equivalent)
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Figures

Videos

Literature Cited

Literature Cited
   Enrietto, P.J., Payne, L.N., and Hayman, M.J. 1983. A recovered avian myelocytomatosis virus that induces lymphomas in chickens. Pathogenic properties and their molecular basis. Cell 35:369‐379.
   Gross‐Bellard, M., Oudet, P., and Chambon, P. 1973. Isolation of high‐molecular‐weight DNA from mammalian cells. Eur. J. Biochem. 36:32‐38.
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