Problems with Promega Taq

Jack Kramer 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:
>
>C.Silvia wrote:
>: 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):

	200mM TRIS
	15mM MgCl2 (adjust to optimize)
	250mM KCl
	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.

jack kramer



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