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1. removed media (centrifuge 4℃로 미리 맞춰준다)
2. PBS wash X 2 (volume 2ml 정도)
3. add TRIZOL to dish (1㎖)
4. stand at RT for 5min, scrap and transfer to E. tube
5. add 0.2㎖ of chloroform
6. vortex for 15sec and stand at RT for 2~3min
7. spin down ×12000g at 4℃ for 15min
8. Transfer the aqueous phase(60% of initiate TRIZOL) to new tube
9. add 0.5㎖ of Isopropanol and at RT for 10min (inverting)
10. ×12000g at 4℃ for 10min
11. wash with 70% EtOH
12. ×12000g at 4℃ for 10min
13. Repeat 11,12 (1-2 times)
14. air dry
(*not to let the RNA pellet dry completely - decrease its solubility)
15. Dissolve 0.1% DEPC treated D.W or Dnase,Rnase freewater
16. incubate at 55~60℃ for 10min
17. -20℃ or -80℃ keep
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