In vitro transcription, Ambion's secret?
jhartley at access2.digex.net
Mon Jul 10 05:11:36 EST 1995
Have you tried pyrophatase?
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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