Phenol/Chloroform?Precipitation?of?DNA

1. Add an equal volume (equal to sample volume) of P/C to sample.

2. Mix (shake, don't vortex).

3. Take aqueous (upper) layer. (If dirty sample, repeat Ph/Chl step until interface is fairly clean).

4. Add equal volume chloroform, mix (shake, don't vortex).

5. Spin 3 min.

6. Take aqueous (upper) layer. (Optional: repeat Chloroform steps).

7. Add 1/10 volume 3M NaOAc (-> 0.3M), mix (shake).

8. Add 2 volumes ice-cold EtOH (100%), mix (shake).

9. Incubate on ice for 15 to 30 minutes. (Can store on ice or at -20°ree;C at this step).

10. Spin 10 minutes, 4°ree;C.

11. Remove supernatant, being careful not to disturb pellet (DNA).

12. Half-fill tube with 70% EtOH, spin at least 2 minutes at 4°ree;C. (Opt: Re-rinse and spin again).

13. Pipet off the sup, careful not to touch pellet.

14. Air-dry (~ 1 hour).

15. Redissolve in 10 mM Tris.