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