Does NaI inhibit T3 RNA Polymerase?

IO00865 at MAINE.MAINE.EDU IO00865 at MAINE.MAINE.EDU
Tue Jun 9 12:31:39 EST 1992


I've been trying to do in vitro transcription reactions using T3 RNA
polymerase to make strand specific probes. I have not been able to get
these reactions to work. Reactions run in parallel using T7 polymerase
work fine. I have tried several preparations of one template and one
try with a different template. I have tried a different batch of enzyme.
I have tried various buffers including one which works for T7 and the one
recommended by the manufacturer. The only thing I can think of is that
trace quantities of NaI (from spinbind, which is essentially the same as
glassmilk) are inhibiting the enzyme. The reactions involving T7 have
been prepared in the same fashion, so whatever it is must be specifically
inhibiting T3 and not T7.
Has anyone had similar problems? Do you have any suggestions for how to
get this reaction to work? Any help will be appreciated.



More information about the Methods mailing list