in vitro tranlation of retroviral gene


Mon Jul 24 15:16:56 EST 1995


In article <Pine.A32.3.91.950724075758.37840A-100000 at umabnet.ab.umd.edu>,
"Mary P. Remington" <mremingt at umabnet.ab.umd.edu> wrote:
> 
> We are interested in the invitro tranlation of a retroviral gag/pol 
> gene.  We are currently trying the Retic Lysate IVT by Ambion.  I am 
> wondering if anyone has experience doing in vitro translation.  Did you 
> use Retic Lysate, Wheat Germ, or Untreated Retic Lysate?  Any opinions, 
> suggestions etc. appreciated.  Regards, Mary

Mary,
We have found that treated Reticulocyte Lysate from Promega works the best
for our applications.  Sometimes we have used wheat germ extract for
different reasons (mainly because of the differences in types of protein
found versus ret. lys.), but have found it to be unreliable and difficult
to obtain consistant results.  We have tried the Ambion lysate, but found
much higher background.  Hope this helps

Darren Boehning 
boehnin1 at jeflin.tju.edu
Thomas Jefferson University

Note: In no way endorsing Promega, my opinion only, ect., ect., ect.



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