quikchange question

Alexander Kraev kraev at bc.biol.ethz.ch
Fri Mar 21 08:44:07 EST 1997


In article <5grpjp$bhg at eldborg.rhi.hi.is>, thorarb at rhi.hi.is (Thorarinn
Blondal) wrote:

> I'm curious about quikchange mutagenesis kit from stratagene.
> has anyone experince concerning the kit?  Is it as fast and accurate 
> as advertized?  What is the max size of plasmid it can mutate?
> 

In our lab many people have used a similar kit from Pharmacia (aka Stratagene's
Chamaleon), and results range from excellent to unusable. The reason for that
 I think is in the design of the mutagenic primer.   Sometimes, it was
just  "calculated"
 by eyeballing. Sometimes, it was designed by a program without sufficient
"fuzzy logic"
 blended in, and apparently did not anneal, which could later be traced to
the secondary 
structure potential at the site of annealing. However, the available
statistics does
not point to a clear set of rules. As you always have to gamble with the
target sequence,
the companies can use it to make "sureshot" kits, which do work with the
positive
control included, but no one can sue a company for his bad luck.

I hope this helps,

-- 
Alexander Kraev, PhD
Biochemie III, ETHZ Zurich
Phone 41-1-632-31-47
Fax 41-1-632-12-13
e-mail kraev at bc.biol.ethz.ch



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