Site-Directed Mutagenesis Kits

John K. Troyer jtroyer at
Mon Sep 18 12:04:27 EST 1995

On Fri, 15 Sep 1995, Jami Grossfield wrote:

> Hi- I am trying to do site-directed mutagenesis and thought using a 
> commercially availbale kit would make this easier. I ordered the 
> stratagene ExSite kit- and their positive control doesn't even work! (yes 
> I have tried varying parameters, tech. support, etc.)
> 1) has anyone else had a problem with this kit?
> 2)does anyone know a good kit for deletion and point mutatation 
> site-directed mutagenesis kits? one they have used??
> I know clontech makes one, and NEB- but I would like input from users 
> before buying another kit! please let me know if anyonw out there has 
> successfully used the ExSite kit from stratagene too!
> thanks!


I have been using the Promega kit (Altered Sites II) with good success.  
In my experience, the major advantages of this kit are:

1) You can introduce multiple mutations simultaneously into the same cDNA
(I have introduced 3 mutations into one cDNA at the same time)

2) The kit works fine with double stranded DNA which greatly simplifies 
the method since you do not have to make a single stranded phagemid

3) This kit is not PCR based.  Therefore, you do not have to worry about 
the fidelity problems inherent in many PCR methods.

Good luck!



John K. Troyer, PhD
University of Maryland School of Medicine
Department of Biological Chemistry
(410) 706-7518
jtroyer at

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