bourson@jouy.inra.fr (Claude Bourson) (03/28/91)
RE:RNAse protection
I generaly use the methods of RNAse protection recomanded in :
Sambrook, Fritch et Maniatis "Molecular cloning" 2nd edition
1989 Cold Spring Harbor Laboratory Press (chapter 7-8)
Ausubel F.M. et al Current Protocol in Molecular Biology
1987-88 Greene Publishing Associated and Wiley-Interscience New-York
(chapter 4)
The methods can be resumed in :
always wear gloves and never use non-treated material.
all the solutions must be treated by 0.2% DEPC (Diethyl pyrocarbonate)
and then autoclaved.(Tris is precipited by DEPC treatment)
glassware is sterilised by backing at 160!C for several hours.
single-use sterilised plasticware is very useful to avoid RNAse problems.
plastic and glass ware can be by traeted with 3% H2O2, or chloroform.
you can also use RNAse inhibitor to increase the level of RNAse
protection.
Good luck.