Phenol/Chloroform Precipitation of DNA
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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.
