Can I use restriction enzymes with human clones in pDNR-Dual?
A. The human pDNR-Dual master clones from HIP were generated using the InFusion reaction (Clontech) rather than an RE approach and most of the RE sites in the MCS are gone after the insert is introduced. In the resulting master clones, the HindIII and SalI sites are present so that enzyme pair can be used to liberate the insert — but beware! There could be HindIII or SalI sites in the insert. See more on the flexibility of using the recombinational cloning approach rather than REs below.
