NZY-A PCR cloning kit

NZY-A PCR cloning kit was designed to allow the direct cloning of PCR products with 3´-A overhangs, which result from amplifications using non-proofreading DNA polymerases such as NZYTaq II DNA polymerase. The cloning vector was prepared by cutting NZYtech´s pNZY28 with EcoR V and adding a 3´ terminal thymidine at both ends. These single 3´-T overhangs improve the efficiency of ligation of a PCR product into the plasmid by preventing re-circularization of the vector and providing compatible overhangs for PCR products generated by non-proofreading thermostable polymerases. Vector pNZY28 contains multiple restriction sites within the multiple cloning region. However, vector digestion with EcoR I or BamH I allows the release of the PCR product since the vector cloning region is flanked by recognition sites of both enzymes. NZY-A PCR cloning system was developed to allow a rapid ligation (1 hour) between the vector and the PCR fragment.