High pure RNA isolation Kit, Roche, Cat.No. 1 828 665


DNase(2 bottles) + Elution buffer  2->5µl dilution

10µl aliguote, -20 store


Isolation of total RNA from cultured cells


1. Resuspend cells in 200µl PBS

2. 400µl lysis - binding buffer & mix well

3. Combine filter tube and the collection tube

4. Sample filter tube ִ´.

5. Centrifuge 15min ,10.000rpm discard the flow-through, again combine

6. 90µl DNase incubator buffer + 10µl DNase mix well, filter tube ְ,   incubate, 5min at RT

7. Add 500µl wash buffer centrifuge, 15sec, 10.000rpm

   Discard the flow-through, again combine

8. Add 500µl wash buffer centrifuge, 15sec, 10.000rpm

   Discard the flow-through, again combine

9. Add 200µl wash buffer centrifuge, 2min at max speed

10. Discard the collection tube. 1.5ml tube غ

11. 50-100µl elution buffer filter tube ְ centrifuge 1min, 10.000rpm.