DNase treatment of RNA sample

Jim Owens jow at helix.nih.gov
Thu Mar 16 10:43:28 EST 1995


In article <199503141158.UAA02322 at bekkoame.bekkoame.or.jp> Toshiharu
Ishizuka, tishizuk at AQUARIUS.BEKKOAME.OR.JP writes:
>        How can I prepare Tris-based DNase buffer without RNase ? I heard
>that diethylpyrocarbonate reacts with Tris, and it is difficult to
>inactivate RNase by autoclaving. 

Try opening a new bottle of high quality Tris.  After making the buffer,
we put them through Nalgene filter units with cellulose nitrate
solutions.  Our group assumes that "ultra pure" Tris has little enough
RNase activity before the bottle is opened that the cellulose nitrate
filter will remove it.  Of course we treat the water used to make the
buffers with diethyl pyrocarbonate before dissolving the Tris.  We have
no data except that our Tris buffers are suitable for use with RNA.  

Wear latex or vinyl gloves on your hands whenever you handle _ANY_
reagent for RNA work or _ANY_ container that will be used with a reagent
for RNA or for solutions of RNA.  The more paranoid you behave, the fewer
problems you will have with RNase.

Good luck,

Jim Owens



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