Problems with Promega Taq
Paul N Hengen
pnh at fcsparc6.ncifcrf.gov
Tue Jun 8 16:58:31 EST 1993
Duncan at genesys.demon.co.uk 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.
>>: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!
>As a producer I have tried various buffers but cannot find one that satisfies
>everyone. Some people swear by the Promega recipe, other prefer Cetus. I can
>give the same enzyme to several labs they only get it to work in their hands
>with a certain buffer. I'm convinced that this is due to the source of
>dNTP's/water/DNA/primers etc they use....[rest deleted]
I have to say I'm behind Jim all the way with this one :-) May I suggest
you work out the conditions for your own personal project with your favorite
enzyme and make up your own buffer? If you're really bent on using a kit
for every little thing, you could buy the PCR Optimizer Kit from Invitrogen
and do the blind color coding scheme so you don't have to even think about
what the components are.
Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA
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