@#$@#% KITS (T7 transcription)
coady at ERE.UMontreal.CA
Thu Oct 21 12:28:36 EST 1993
In article <CF9A0p.CpJ at usenet.ucs.indiana.edu> jgraham at bronze.ucs.indiana.edu (the End) writes:
>The day I feared has predictably come. Current advances in molecular
>biology techniques are no longer aviavlable to the general science
>public via the literature, but are being sold to us through commerical
>companies at inflated prices.
>Case in point, T7 transcription. Ambion and Epicenter sell kits that
>show 10-50 times better yields of RNA product from 1 ug of DNA template.
[Deletions here... no frameshifts, though :-) )
>Ambion pamphlet which indicated that a key feature is an increase in
>NTP substrate concentration from 0.5 mM to 7.5 mM , " a level normal
>inhibitory" to polymerase function. Their reaction conditions which
>prevent inhibition actually have a patent pending.
i [More deletions ]
>Any comments on how elevated NTP levels might be used without inhibition
>(eg. Mg++ chelation ?) or any guesses on the contents of these kits
>or other information on increasing yields most appreciated.
The results in their ads as well as the description that you gave
are similar to an article that was published a while before the kits
became available. Check out Gurevich et al, Anal. Biochem. vol. 195,
pp. 207-213 (1991) "Preparative in vitro mRNA Synthesis Using SP6 and
T7 RNA Polymerases."
I hope this helps.
Michael J. Coady
COADY at ERE.UMONTREAL.CA
"Louis, I think this could be the start of a beautiful .sig file..."
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