SP6 RNA polymerase -- HELP

Walter Blank wblank at gpu.srv.ualberta.ca
Thu Sep 7 12:59:14 EST 1995


Is there some kind of trick to using this stuff?

Doing a side-by-side transcription with T7 RNApol with a fresh batch of
enzyme under exactly the same conditions (same vector, NTP's, buffer (as
recommended by Pharmacia), etc.) I get about 1% of the yield from SP6 as
from T7.

Will doing the reaction at 40C help a lot? Also, I'm using fairly high SA
32P-UTP (3000 Ci/mmol, 10uCi/25uL reaction) -- Does the SP6 pol require a
higher concentration than T7?

Any hints are more than welcome.  Thanks bunches.

Wally

Sei immer freundlich.  It's all much appreciated.  Out!



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