Problems with Promega Taq
Vivian Miao
VMiao at oregon.uoregon.edu
Thu Jun 10 17:26:22 EST 1993
In article <1v57hjINNio6 at lynx.unm.edu>, bhjelle at carina.unm.edu () wrote:
> In article <Miao-030693075250 at fp1-molbio-13.uoregon.edu> Miao at oregon.uoregon.edu (Vivian Miao) writes:
>> side by side tests with NEB's Vent... for my setup,
>>Taq makes a lot more product than Vent
>>the reactions are made up as a cocktail minus
^^^^^^^^^^^^^^
>>enzyme and its conc'd buffer stock which are added later, the
^^^^^^^
> We normally make sure there are some salts in the
> reaction mix before we add the enzyme, on the assumption
> that diluted enzyme may denature in a situation where
> there is no salt or buffer. Maybe all you are seeing is
> a differential sensitivity to denaturation of Vent compared
> with Taq.
>
> Brian
sorry, bad editing on my part in the original post
First cocktail = template, oligos, NTP's, water.
Then salts,
Then enzyme
(Then oil)
(Then take bets on what will go wrong this time, etc. ...:-)
Vivian
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