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
