Department of Biological Sciences,

1000 Hilltop Circle

Baltimore, MD 21250

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. (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)

This Web page is maintained by Julie B. Wolf, UMBC;
Last updated on 3/2/2010