TRIzol method for RNA isolation

Chen-Ming Fan Lab,Carnegie Institute of Washington

TRIzol method for RNA isolation
-To tissue or cells, add 200- 1 ml TRIzol (GIBCO BRL)
- homogenize for 60 sec (vortex or polytron)
- add 40ul-200 ml chloroform (1/5 volume of Trizol)
- mix by vortexing for 15 sec
- incubate for 5 min at room temperature
- centrifuge at 12.000 g for 15 min
- transfer the aqueous phase into a new tube
- add 500 ml isopropanol
- centrifuge at max for 10 min in the cold room
- wash the pellet with 500 ml 70 % ethanol
- centrifuge at max for 2 min in the cold room
- air-dry the pellet for 10 min
- disolve the pellet in 50-100 ml DEPC-H2O