Nucleases inhibitors

Antonio Rodriguez Franco bb1rofra at uco.es
Thu Sep 14 06:32:05 EST 2000


I am trying to run NASBA amplification reactions, and have got strong
evidences indicating that both RNAses and DNAses are present in my
extract in high quantities.

NASBA implies the amplification of RNA by using Transcriptase inverse,
T7 RNA polymerase and RNAse H

I can get rid of problems with RNAses using the placental inhibitor.
Amersham claims in its booklet that this inhibitor does not affect
either of these 3 required activities.

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)






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