- Harvested cells can be stored in -80 degree Celsius for less than a week before RNA isolation or right after harvest.
- Add 400 µl Trizol to lyse the cell and incubate at Room Temperature for 5, minutes.
- Add 80 µl chloroform (0.2ml per 1ml Trizol)
- Vortex the mixture for 15 secs and incubate at Room Temperature for 10 minutes
- Centrifuge at 12000rpm / 4 degree Celsius / 15 minutes
- Transfer the aqueous phase to the new clean tube and add equal volume of isopropanol
- Mix well (short vortex)
- Incubate at room temperature for 10 minutes.
- Centrifuge at 12000 rpm / 10 minutes/ 4 degree Celsius
- Remove isopropanol using pipette keeping the pellet safe
- Add 400 µl of 75% ethanol and centrifuge at 7500 rpm/ 5 minutes/ 4 degree Celsius
- Pipette out the ethanol keeping the pellet safe
- Air dry the pellets for 5-10 minutes
- Resuspend the pellet with 20 µl Nuclease Free water.
- Measure the concentration of RNA isolated. (SPECTROPHOTOMETRIC ANALYSIS)
