RNAse protection

Claude Bourson bourson at jouy.inra.fr
Thu Mar 28 09:33:02 EST 1991


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.



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