The making of a DNA construct with deletions?
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
>> by amplifying the different cassettes of DNA with flanking restrictions
>> sites and then joining them, or is there another way to get around? Any
>> 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
Thanks for the references and the hints on how to do this. I think that I
will try the method described in Biotech.
More information about the Methods