primer design for stratagene's quick change mutagenesis

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Tue Aug 24 05:24:31 EST 1999



Frederik Boernke wrote:

> Another Question: Is it really necessary to buy this as a kit, I mean
> all components are regular PCR reagents/restriction enzymes. We use
> Turbo Pfu on a regular base and also have some DpnI in the freezer.
> To use the things we've got to me seems a much cheaper alternativ to
> the kit, or does it contain anything special that makes it
> indispensable.
>
> Ricky

Hi Ricky,
That is a question that often comes to mind when a kit-based method is
used. The answer is always the same:
OF COURSE you can do everthing on your own, using the materials you have
and are used to. Kits are convenient when you perform a method only a
few times, because then they may in fact be cheaper than buying all
components separately. In addition, the kit components are usually
tested for exactly the method they are sold for, so you can be sure they
work even with little lab practice. Thus, there are reasons to buy kits,
but I have seen  that kits are more and more used in places where they
should not. In particular, the training of lab novices should be free
from kits. Rather, conventional methods should be trained there because
they normally give insights not obtainable from kits, and strengthen
peoples abilities to do unconventional, spontaneous, and often NOVEL
things.
As to the quickchange kit: It is convenient to buy it if you don´t have
the enzymes and high-quality competent bacteria handy. Otherwise you can
do the mutagenesis with the available materials. I have done so several
times, and of course it worked in the same way as the kit. Anyway, I
have to admit that I still use the kit because it is VERY convenient to
have everything in a single box and dedicated to only one purpose. I
also find the price reasonable.


Frank




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