>I am curious to hear what is the longest insert anyone has cloned
>using a modification of the Quikchange cloning strategy. Basically,
>ligation-independent cloning by strapping on homologous regions of
>the vector onto the primers which also generate the initial PCR
>product. I plan to proceed with my insert which is ~ 2kb and am
>curious to get some feedback if you have successfully cloned inserts
> > 1.5kb using the above strategy.
Here, the maximum we've tried was 3 kbp and it worked just as well as with
the the smaller fragments. Most of the Quickchange cloning we did involved
1500-300 bp and no obvious differences in success rate vs size come to mind.
Dima
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