The three tools for doing Gateway cloning in the CLC Main Workbench mimic the procedure followed in the lab:
- First, attB sites are added to a sequence fragment
- Second, the attB-flanked fragment is recombined into a donor vector (the BP reaction) to construct an entry clone
- Finally, the target fragment from the entry clone is recombined into an expression vector (the LR reaction) to construct an expression clone. For Multi-site gateway cloning, multiple entry clones can be created that can recombine in the LR reaction.
For more information about the Gateway technology, please visit http://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cloning/Gateway-Cloning.html
To perform these analyses in the CLC Main Workbench, you need to import donor and expression vectors. These can be downloaded from Invitrogen's web site and directly imported into the Workbench: http://tools.invitrogen.com/downloads/Gateway%20vectors.ma4