NZY-A Speedy PCR cloning kit

NZY-A Speedy PCR cloning kit was designed to carry out fast andefficient cloning of PCR products with 3'-A overhangs, which resultfrom amplifications using non-proofreading DNA polymerases suchas NZYTaq II DNA polymerase. This methodology combines theefficiency of an improved ligation buffer with the speed of SpeedyLigase to allow a rapid ligation between the vector and the PCRproduct in only 5 minutes at room temperature (18-25 °C). Thecloning vector was prepared by cutting NZYTech's pNZY28 withEcoR V and adding a 3' terminal thymidine at both ends. Thesesingle 3'-T overhangs improve the efficiency of ligation of a PCRproduct into plasmids by preventing re-circularization of the vectorand providing compatible overhangs for PCR products generated bynon-proofreading thermostable polymerases. Vector pNZY28contains multiple restriction sites within the multiple cloningregion. However, vector digestion with EcoR I or BamH I allows therelease of the PCR product since the vector cloning region is flankedby recognition sites of both enzymes. Components: * NZY-A Speedy Buffer (4x) * pNZY28-A vector (50 ng/µL) * Speedy Ligase (5 U/µL) * NZY-A positive control insert (50 ng/µL) * NZYStar Competent Cells (*) * Competent Cells Control Plasmid (0.1 ng/µL) (*)(*) only provided in MB13701 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, Fast. Product Category: DNA Cloning & Mutagenesis. Protocol Time: Approx. 10 min