Phenol/chloroform Extraction of DNA
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620
Materials:
 |
phenol:chloroform (1:1) |
|
|
chloroform |
- Add an equal volume of buffer-saturated phenol:chloroform (1:1) to the DNA solution.
- 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 )
- Spin in a microfuge for 3 min.
- 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)
- To remove traces of phenol, add an equal volume of chloroform to the aqueous layer.
- Spin in a microfuge for 3 min.
- Remove aqueous layer to new tube.
- Ethanol precipitate the DNA
