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QuikChange™ Site-Directed Mutagenesis Kit - (Aug/28/2007 )

i have a Question for PRIMER DESIGN in QuikChange™ Site-Directed
Mutagenesis Kit

when i want to insert 4 amino acid the really Tm should contain the 4 amino acid or not

and if any one has experience in this kit let tell us who he can use it

thank you

-biojoe-

QUOTE (biojoe @ Aug 28 2007, 02:33 PM)
i have a Question for PRIMER DESIGN in QuikChange™ Site-Directed
Mutagenesis Kit

when i want to insert 4 amino acid the really Tm should contain the 4 amino acid or not

and if any one has experience in this kit let tell us who he can use it

thank you


The Tm should only be calculated on the bases that will anneal to the template, so you shouldn't include the bases you are mutagenising.

-bitesizebio guy-

Hi,

Stratagene has a nice website on site-directed mutagenesis. They also have an online oligo design program, which you may find useful. You'll have to register for an account (takes about 10 seconds) but will have access to their manuals.

http://stratagene.com/products/showCategory.aspx?catId=190

Good luck!

-NPMALK-

underprocess

-biojoe-

Hello friends

I did what the say in the kit but I have a problem when I get my amplified pcr I digested with Dpn1 after that I transform in competent cells after that I can't have any colony in the petridish and I’m sure a bout my competent cell I did (+) control and I have the colony so I’m really surprise

Do any one had opinion for this situation

-biojoe-