The making of a DNA construct with deletions?

as nospam at nospam.com
Tue Mar 7 06:36:37 EST 2006


<troels.wind at gmail.com> wrote in message 
news:1141381892.884857.115350 at p10g2000cwp.googlegroups.com...
>
> as skrev:
>
>> Dear NG.
>> I have a 1500bp DNA construct where I want to delete 5-6 smaller regions
>> dispersed throughout the sequence. What is the easiest way to do this? Is 
>> it
>> by amplifying the different cassettes of DNA with flanking restrictions
>> sites and then joining them, or is there another way to get around? Any 
>> help
>> would be appreciated. Thanks.
>
> You could try the approach described in BioTechniques 29:970-972 (Nov
> 2000). I haven't tried this myself, but a collegue has had some luck
> with it. It's basically a QuickChange reaction where the portion to be
> deleted from the template is looped out by the mutagenesis primers. One
> strange observation in the paper is, that only one mutagenesis primer
> is needed, not the two complementary ones normally used for
> QuickChange. How this works is a mystery to me...
>
> /Troels, University of Aarhus, Denmark

Hi everyone
Thanks for the references and the hints on how to do this. I think that I 
will try the method described in Biotech.
Thanks.
A.Salanti 




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