Golden Gate Cloning Technique

Golden Gate cloning is a standardized cloning method for the directional assembly of DNA that allows the combination of multiple DNA fragments using a single reaction. ​Golden Gate technology is made possible by the special abilities of type IIS restriction enzymes that can cleave DNA outside of their typical recognition sites. The recognition sites for type IIS restriction enzymes can be engineered to produce desired DNA overhangs, allowing scientists to build and seamlessly litigate DNA fragments. Using this restriction and litigation technique researchers achieve a high level of construct assembly, approximately 95-100% of colonies generated contains the desired DNA construct.