Purification and Concentration of DNA from Aqueous Solutions

David Moore1

1 Baylor College of Medicine, Houston
Publication Name:  Current Protocols in Protein Science
Unit Number:  Appendix 4E
DOI:  10.1002/0471140864.psa04es13
Online Posting Date:  May, 2001
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Abstract

This unit presents basic procedures for manipulating solutions of single‐ or double‐stranded DNA through purification and concentration steps. Phenol extraction with ethanol precipitation is described along with an alternate procedure of precipitating DNA with isopropanol. A describes concentration of DNA using butanol.

     
 
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Table of Contents

  • Basic Protocol 1: Phenol Extraction and Ethanol Precipitation of DNA
  • Alternate Protocol 1: Precipitation of DNA Using Isopropanol
  • Support Protocol 1: Concentration of DNA Using Butanol
  • Literature Cited
     
 
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Materials

Basic Protocol 1: Phenol Extraction and Ethanol Precipitation of DNA

  Materials
  • DNA to be purified (≤1 mg/ml) in 0.1 to 0.4 ml volume
  • 25:24:1 (v/v/v) phenol/chloroform/isoamyl alcohol (made with buffered phenol ; appendix 2E)
  • 3 M sodium acetate, pH 5.2 ( appendix 2E)
  • 100% ethanol, ice cold
  • 70% ethanol, room temperature
  • TE buffer, pH 8.0 ( appendix 2E)
  • Speedvac evaporator (Savant)

Alternate Protocol 1: Precipitation of DNA Using Isopropanol

  • sec‐butanol
  • 25:24:1 phenol/chloroform/isoamyl alcohol (made with buffered phenol ; appendix 2E)
  • Polypropylene tube
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Figures

Videos

Literature Cited

   Marmur, J. 1961. A procedure for the isolation of desoxyribonucleic acids from microorganisms. J. Mol. Biol. 3:208‐218.
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