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Phenol/chloroform Extraction of DNA


bullet phenol:chloroform (1:1)
bullet chloroform
  1. Add an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution.
  2. Mix well. Most DNA solutions can be vortexed for 10 sec except for high molecular weight DNA which should be gently rocked. (If using Phase-Lock Gel, follow procedure M.1)
  3. Spin in a microfuge for 3 min.
  4. Carefully remove the aqueous layer to a new tube, being careful to avoid the interface. (Steps 1-4 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 min.
  7. Remove aqueous layer to new tube.
  8. Ethanol precipitate the DNA( see ethanol precipitation)

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Last updated on 3/2/2010

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