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.