TRIzol method for RNA isolation

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