NZY-A PCR cloning kit

NZY-A PCR cloning kit was designed to allow the direct cloning of PCR products containing 3´-A overhangs, resulting from amplifications using non-proofreading DNA polymerases, such as Taq DNA polymerase. The cloning vector provided in the kit was prepared by cutting NZYTech´s pNZY28 with EcoRV 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 EcoRI or BamHI 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.