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 |
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