TNT T7 kit (Promega) with (Stratagene) SP6 pol

rompubil from gmail.com via methods%40net.bio.net (by rompubil from gmail.com)
Fri Feb 6 04:15:32 EST 2009


Hello,
My lab has the TNT T7 Coupled Reticulucyte Lysate System (Promega).
However the cDNA I want to transcribe/translate is in a plasmid under
the control of Sp6 promoter. My lab has the Sp6 RNA polymerase from
Stratagene. Therefore, has somebody try to add to the TNT mixture a
different RNA polymerase (from a different company)? I have found some
posts describing a buffert that can be used in that kit. However,
these posts were related to T7 RNA polymerase.

I would prefer to avoid subcloning the cDNA in a different plasmid and/
or carrying out transcription/translation in two steps.

Thanks
Rom


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