mutagenesis kit

David L. Haviland david.l.haviland at uth.tmc.edu
Thu Sep 6 12:57:25 EST 2001


----- Original Message -----
From: "Wolfgang Schechinger" <wolfsc at ibms.sinica.edu.tw>
To: <methods at hgmp.mrc.ac.uk>
Sent: Wednesday, September 05, 2001 11:38
Subject: Re: mutagenesis kit


> Dear Bob,
>
> I also observed this sometimes. Any ideas why? Sort of
> incorporated primer dimer?

Wolfgang:

I once chatted with a Stratagene rep and they are well aware of the problem.
The going thoughts were that you've got oligos hybridizing to each other and
to parts of each other as they are both in the reaction and complementary.
Stratagenes view was that you need to sequence all clones (no surprise) but
as long as you get one with what you want, "so what" if you get a handful
with primer dimer incorporation?  In any case, the system works and works
well for us.   However, we were never able to substitute the "kit" and
thought there was something "magical" or proprietary in the buffer or dNTPs
(we tried 200uM) that enabled it to work.

David

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