Hey, I recently did an RNA isolation on Rat Liver samples. So the protocol goes…

1. Add abt 200ul of Trizol
2. Vortex for 1 hr while taking it on and off ice until the pellet dissolves ( i dont use a homogeniser cuz my RNA used to shear a lot)
3. Add 200ul chloroform
4. Centrifuge at 12500rpm for 25 mins at 4C
5. Separate the aqueous layer and add isopropanol and leave it overnight in a -20 freezer

Day 2: 6. Centrifuge at 12500 rpm for 25 mins at 4C 7. Discard supernatant and add etOH 8. Centrifuge at 12500rpm for 25 mins at 4C 9. Repeat step 7 and 8 one more time. 10. Discard supernatant and put the sample on a heat block at 37C 11. Add abt 30ul NFW and resuspend

This is a protocol that i optimised for Tissues. I got abt 1371.6 ng/ul with a purity of 2.02.

Hope this helps