Problems with Promega Taq
kramer at oj.rsmas.miami.edu
Wed Jun 9 14:35:09 EST 1993
In article <122014 at bu.edu> pfoster at bu.bu.edu (Patricia Foster) writes:
>: Promega's Taq doesn't like Cetus buffer, and the presence of detergent in
>: the Promega buffer may be the reason, altho I haven't tried to add
>: detergent to the Cetus stuff.
>Jack Kramer wrote:
>:I had similar problems with the Promega Taq with <Promega's 10X
>:buffer. The Cetus Taq had similar problems in the same buffer.
>:Switching to the Cetus buffer cured the problem. With this buffer,
>:or modifications of it which <i make up myself now, I get identical
>:results with either Taq. Try Promega Taq in storage buffer B and make
>:up your own reaction buffers - saves bundles of money!
>Huh? Could we have the recipes?
I have found the following to give identical results for both Cetus and
Promega Taq (storage buffer B):
15mM MgCl2 (adjust to optimize)
0.1% Gelatin (critical to get identical results)
0.5% tween 20
I thaw and dilute the enzyme as received into this same buffer to 1 unit
per microL and store at 4C for use within 2 weeks. Difco BActo gelatin
works fine and is cheap. Other detergents can be used.
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