Brand : NZYTech Description: NZY-A Speedy PCR cloning kit was designed to carry out fast and efficient cloning of PCR products with 3´-A overhangs, which result from amplifications using non-proofreading DNA polymerases such as NZYTaq DNA polymerase. This methodology combines the efficiency of an improved ligation buffer with the speed of Speedy Ligase to allow a rapid ligation between the vector and the PCR product in only 5 minutes at room temperature (18-25 °C). 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 plasmids by preventing recircularization 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.
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