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Description

The pT7Blue(TM)-2 T-Vector is designed for simplified cloning of PCR products. Many thermostable DNA polymerases, such as Taq DNA polymerase, leave single 3' A-nucleotide overhangs on their reaction products. These molecules can be ligated directly to a vector containing compatible single T-nucleotide overhangs. The pT7Blue(TM)-2 vector has been specifically constructed for thisapplication and has been prepared for T-cloning by digestion with EcoRV followed by the addition of single 3' dT residues at each end. The T-Vector is provided ready to ligate with DNA amplified with any DNA polymerase that leaves single 3' dA overhangs. Target inserts having other types of ends, such as reaction products of proofreading polymerase or restriction fragments, can be easily prepared for T-cloning with the Single dA Tailing Kit.The pT7Blue(TM)-2 Vector is a versatile plasmid featuring the 43-bp Xenopus beta-globin 5'-UTR for enhancement of translation in rabbit reticulocyte lysates and frog oocytes. Inserts cloned in-frame with the lacZ alpha-peptide coding sequence can also be expressed in E. coli from the lac promoter. The vector has many additional features that make it an excellent choice for a variety of applications.