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. Components: * NZY-A Buffer (2x) * pNZY28-A vector (50 ng/µL) * T4 DNA Ligase (2 U/µL) * NZY-A positive control insert (50 ng/µL) * NZYStar Competent Cells (*) * Competent Cells Control Plasmid (0.1 ng/µL) (*)(*) only provided in MB05301 kit. Shipping Conditions: Dry Ice. Storage Conditions: Competent cells: -85 °C to -65 °C-Other components: -30 °C to -15 °C. Application: TA Cloning. Features: Blue/White screening. Product Category: DNA Cloning & Mutagenesis. Protocol Time: Approx. 60 min