Phenol/Chloroform Extraction of DNA

Materials
  • Phenol:Chloroform (1:1)
  • Chloroform
Procedure
  1. Add an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution.
  2. Mix well. Most DNA solution can be vortexed for 10 sec except for high molecular weight DNA which should be gently rocked. Spin in a microfuge for 3 min.
  3. Carefully move the aqueous layer to a new tube, being careful to avoid the interface.
  4. Steps 1 to 3 can be repeated until an interface is no longer visible.
  5. To remove traces of phenol, add an equal volume of chloroform to the aqueous layer.
  6. Spin in a microfuge for 3 minutes.
  7. Remove aqueous layer to new tube.
  8. Ethanol precipitate the DNA.

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