solutions to storage oligonucleotides
tk at csail.mit.edu
Thu Nov 4 00:56:50 EST 2004
fabio <fabio at fabius.it> writes:
> becouse my routine PCRs are usualy RT-PCR and someone is of the type
> one-step, I used to prefer water RNAse free (becouse treated with DEPC
> and auocaved two times -two to be sure of destroy all DEPC), but I
> know that DEPC demage Tris and, in the other hand the DEPC change the
> pH of water if I remeber well, and the change is added to the one of
> the resin "milli-Q" in the direction of acidity.
> Work well without DEPC, even if good, isn't a garanty to don't have
> RNAase, but maybe there are not choice.
> What is your opinion?
Typically Tris buffers for RNA work are made with DEPC treated water
and solid tris. An alternative is to buy Tris buffers which are RNAse
free. These are available at final concentrations and at 1M, which
can be diluted with DEPC treated or other high quality water.
Also, high quality DI water systems with recirculation and UV
treatment produce RNAse free water without treatment. You have to be
careful that your solid Tris is not contaminated with sloppy
technique. You can also buy HPLC/reagent grade nuclease free water
and make solutions with that.
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