Nucleases inhibitors

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Thu Sep 14 09:45:31 EST 2000


In article <39c0acee.96157357 at news.cica.es>, Antonio Rodriguez Franco
<bb1rofra at uco.es> writes
>Now, I would like to be sure that DNAse activity is gone as well. I
>cannot boil my samples, because I use RNA from the starting point.  I
>also need Mg ions in the reaction, son cannot use EDTA. And not know
>of any known inhibitor I can use for this purpose.
>
>Any hint?
>(forget about purifications, only interested in inhibitors)

I have doubts that you are going to find a DNase inhibitor let alone one
that will not affect one or more of the three enzyme components needed.

I know you don't want to talk about purifications but that is the best
place to start for removing those activities. Whatever purification
method you are using for your RNA you really need to make sure that,
that is kiilling off those activities. 

Can you be sure that your NASBA reaction components themselves are free
of DNase and RNase contamination.

On a side note are you using homemade NASBA mixes or commercial. Just
curious as I want to give NASBA a go myself.

Duncan
-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
FAX: +44(0)8701640382
http://www.dnamp.com
http://www.genesys.demon.co.uk






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