University of Pennsylvania
DNA Sequencing Facility
Cloning and Sub-cloning
Cloning strategy is designed jointly by the user and the facility. Directional or non-directional cloning into any vector is performed by including right restriction sites in the primers, if necessary, for amplifying the insert for PCR-based cloning. After cloning the insertions sites are checked and orientations are determined by either restriction digestion and/or sequencing.
PCR products are cloned using Invitrogen's TA cloning system or any other vector as requested.
After designing the cloning strategy, the insert is retrieved from the donor vector and appropriate restriction sites are created if necessary. The insert is then cloned into appropriate recipient vector followed by the screening for correct clones by restriction digestion and/or sequencing.
We also offer ligation-independent cloning (LIC) when requested by the user.
The DNA Sequencing Facility is an Abramson Cancer Center Shared Resource that is approved and partially funded by the National Cancer Institute.