Joyce: Phenol extraction

Concept

• To remove contaminating protein that could inhibit downstream protocols, or affect the quality of your DNA in the long-term.
• Phenol completely denatures proteins and concentrates them in the nonpolar (phenol) phase, leaving DNA undamaged in the aqueous phase

Procedure

1. In a fume hood and while wearing gloves, add an equal volume of TE-saturated phenol to your DNA sample WARNING: Wear gloves and do this in a fume hood; phenol is some nasty stuff
2. Vortex for at least 1 minute
3. Spin at 14K RPM for 2 minutes
4. Pipette off the aqueous phase to a new tube
5. Add an equal volume of water-saturated ether to the DNA solution (fume hood, wear gloves)
6. Vortex for at least 1 minute
7. Spin at 14K RPM for 2 minutes
8. Pipette off the ether phase (ether is on top), discard in a falcon tube containing activated charcoal in the fume hood
9. Repeat the ether wash again
10. [Joyce:DNA precipitation|Precipitate the DNA]