In vitro transcription, Ambion's secret?

jim hartley jhartley at access2.digex.net
Mon Jul 10 05:11:36 EST 1995


Have you tried pyrophatase?

Jim Hartley

On 6 Jul 1995, Eric A. First wrote:

> Hi netters,
>         We're trying to set up large scale in vitro transcription reactions
> to synthesize tRNA. We've used Ambion's T7-MEGAshortscript kit with good
> results.  We've also used a homemade kit with less than impressive results.
> Swapping various parts between the kits tells us that the main difference
> is in the enzyme mix used by Ambion (our homemade enzyme mix contains T7
> RNA polymerase, RNasin, and acetylated BSA).  Does anyone know what Ambion
> puts in their enzyme mix that makes it work so well (i.e. is there a
> literature reference)?  We've tried adding as much as 50 units of T7 RNA
> polymerase per 20 ul reaction and still get a 10-fold lower yield than with
> the Ambion enzyme mix.  Any ideas or suggestions on how we can increase our
> yield?  Thanks.
> 
> Eric First (efirst at lsumc.edu)
> 
> 
> 
> 



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