Phenol Chloroform Extraction ETOH Precipitation[Isolation]

1. Bring DNA solution volume up to 100uL in water
2. Add 100uL of TE saturated phenol (bottom layer) and vortex 15-30s
3. Add 100uL of Chloroform and vortex
4. Spin max speed for 5min at room temp
5. Remove aqueous phase (top layer) carefully and place ~90% in a fresh pre-labeled 1.5mL tube. Note: be careful to avoid the protein layer at the aqueous: phenol interface
6. Add 1/10th volume NaOAc (2M) pH 5.2 or 5.4
7. Add 2.5 volumes (~250uL) of ETOH
8. Place tubes in -80C freezer for 10min
9. Spin 4C for 20min 1
10. Decant supernatant
11. Wash pellet 2x with 70% ETOH 12 Let pellet airdry and re-suspend in 10uL of water.

February 5, 2007 tj