Site directed mutagenesis

novalidaddress from nurfuerspam.de via methods%40net.bio.net (by novalidaddress from nurfuerspam.de)
Fri Aug 5 16:21:56 EST 2011


Hi Matt, 

no real need for a kit, they are just expensive and possibly a waste of money. Get your mutagenesis primers (at least one) and some DpnI. It might be wise to use a not proofreading enzyme, sequence some of your possible hits and if necessary, subclone a convenient stretch of your target gene. 

You might want to search the archive of this NG for Qui(c)kChange etc, this question comes up every now and then.  

If your sequence is not too long you might want to check if gene synthesis might be an alternative.

HTH & have fun with your mutants!

Wo

-------- Original-Nachricht --------
> Datum: Fri, 5 Aug 2011 18:46:56 +0100
> Von: mnr mnr <mnr475 from gmail.com>
> An: methods from magpie.bio.indiana.edu
> Betreff: Site directed mutagenesis

> Hi all.
> 
> In your experience, what are the pros and cons of using Strategene
> QuikChange and Phusion site-directed mutagenesis kit? Is one better than
> the
> other cost and time wise? I intend to prepare a set of deletion and point
> mutations of my gene.
> 
> Thank you.
> 
> Matt
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> Methods mailing list
> Methods from net.bio.net
> http://www.bio.net/biomail/listinfo/methods

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