RNA Isolation using RNAzol (10cm Dish)

1.   Wash cells 2 X in PBS

2.   Add 1ml RNAzol

3.   Scrape cells and transfer to an eppendorf tube

4.   Add 200µl Chloroform

Vortex 5 seconds

5.   3 minutes @ Room Temperature

6.   CFG 15min @ 10K RPM and 4°C

Transfer 750µl of the Aqueous Þ New Tube

7.   Add 750µl Isopropanol

Vortex 5 seconds

8.   10 minutes @ Room Temperature

9.   CFG 10min @ 10K RPM and 4°C

Pour off

10.        Add 1ml of 95% EtOH

11.        CFG 5min @ 8K RPM and 4°C

Pour off and Air Dry 10 minutes at room temperature

12.        Resuspend in 100 ml of Nuclease free water

13.        Dilute 5ml of RNA with 495ml of water

14.        Read in the Spec