# RNA extraction with TRIzol (invitrogen cat# 15596-026)  
1. Harvest 100 mg tissue (about 1 leaf) and flash freeze in liquid nitrogen  
2. Add 1ml TRIzol  
3. Homogenize and incubate at room temperature for 5 minutes  
4. Add 200μl chloroform and shake for 15 seconds  
5. Incubate 2 minutes at room temperature  
6. Centrifuge 12000 rpm for 15 minutes at 4°C  
7. Transfer aqueous phase to new tube containing 500μl isopropanol  
8. Incubate 10 minutes at room temperature  
9. Centrifuge 12000 rpm for 10 minutes at 4°C  
10. Discard supernatant and wash pellet in 1 ml 75% ethanol (made with DEPC water)  
11. Centrifuge at 7000 rpm for 5 minutes at 4°C  
12. Air dry for 10 minutes  
13. Resuspend in 30μl DEPC water  
14. Incubate 10 minutes at 55°C  
Store at -80°C

# Extra steps to have a cleaner sample  
1. After step 13 add 3μl NaOAc  
2. Add 60μl ethanol 100%  
3. Incubate at -80°C for 30 minutes  
4. Centrifuge at maximum speed for 30 minutes at 4°C  

1L DEPC water  
Add 1ml DEPC to 1L milli-Q water  
Let sit overnight  
Autoclave the next day