grenet_o at a1.tobias.sandoz.com
Tue Jul 2 02:05:41 EST 1996
I'm trying to make an in vitro transcription from a 830bp insert
cloned in pKS Bluscript from Stratagene.
I'm using the pT3 riboprobe system of Promega.
I need a full length RNA, but unfortunatly I obtain 2 of them: The
good one and a shorter one.
I assume that there is a terminator, (hairpin formation), in the
I've plan to clone the insert in another vector, so that I can
use another polymerase...
I tried already to decrease the temperature of the in vitro
transcription reaction down to 250C, (the kit says so...). I tried
also to increase the temperature up to 400C to avoid hairpin
formation, but in the first case I still obtain 2 products, and
in the second one, no product at all.
Can you tell me if you think to something else that can help me
to avoid the new cloning step.
P.S: From the other side of the insert, the pT7 riboprobe system
is working properly.
Please send help to:
GRENET at dstox.pharma.sandoz.ch
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