Midi RNA preparation using Trizol reagent

        
                  
 


1.        Grind ~1.5g arabidopsis seedlings with liquid N2 .
2.        Transfer to falcon tube (15ml/6000G) preadded with 10ml Trizol,
mix well, stand for 5min.
3.        Add 2ml chloroform, stand for 3min.
4.        7000 rpm (JA20, ~6000g) 25min.
5.        Take supernatant (6-7ml), add 7ml IPA.
6.        RT 10min.
7.        7000 rpm 25min.
8.        Wash the RNA pellet with 75% EtOH.
9.        7000 rpm 5min.
10.        Dry
11.        Add 200ul DEPC water.
12.        Mix well, 55C 10min.
13.        7000rpm 5min.
14.        Transfer to 1.5ml eppendorf tube.
15.        -80C.