Plant Laboratory

Total Plant RNA Isolation-Trizol Reagent Method Total Plant RNA Isolation Using Trizol Reagent 1. Crush 100 mg of tissues in 1mL of triazole reagent by pestle mortar or micro pestle 1.5 or 2.0 mL RNase free Eppendorf tubes. *Incubate for 2-3 minutes at room temperature. 2. Add 200 uL chloroform and mix by gently inverting the tube. *Incubate for 2-3 minutes at room temperature. 3. Centrifuge at 12000xg at 4oC for 15 minutes. 4. Transfer the supernatant into fresh 1.5 mL RNA’s free tube. 5. Add 500 μL isopropanol mix gently by inverting the tube. *Incubate for 10 minutes at room temperature. 6. Centrifuge at 12000xg at 4oC for 10 minutes. *Discard the supernatant and keep the pallet in the tube. 7. Add 1mL of ice-chilled 75% ethanol to wash the pallet. 8. Centrifuge at 7500xg at 4oC for 5 minutes. *Discard the supernatant and keep the pallet in the tube. *Keep lid opened tube at 4oC for 15 min to air dry the pallet. 9. Add 50-100 uL RNA’s free water in pallet and