Phenol/Chloroform Extraction of DNA

• Phenol:Chloroform (1:1)
• Chloroform

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.