Site-Directed Mutagenesis Kits

John K. Troyer jtroyer at umabnet.ab.umd.edu
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!



Jami:


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!


Regards,

John


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



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