Gateway™ pDEST™20 Vector - FAQs

Can restriction enzymes be used to linearize pDEST vectors to increase recombination efficiency?

Destination vectors can be linearized by cleaving at a restriction site outside the ccdB gene but within the region of the Gateway cassette. For example, pDEST10 can be linearized at the Mlu I site, while pDEST27, pDEST26, and pDEST20 can be linearized at the EcoR I site. However, please note that although we used to recommend using a linearized destination vector for more efficient recombination, further testing has shown that linearization is usually NOT required to obtain optimal results for an LR recombination reaction.