Phenol/chloroform Extraction of DNA

Materials:

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 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.

5.(Steps 1-4 can be repeated until an interface is no longer visible) .

6.To remove traces of phenol, add an equal volume of chloroform to the aqueous layer.

7.Spin in a microfuge for 3 min.

8.Remove aqueous layer to new tube.

9.Ethanol precipitate the DNA.