Construction of T-tailed vectors derived from a pUC plasmid: a rapid system for direct cloning of unmodified PCR products.
We constructed two new T-vectors called pUCTA119 and pUCTA18, derived from pUC18. The vectors were designed to produce single thymidine (T)-overhangs when digested with a restriction enzyme Eam11051. The use of the vectors provides a very rapid system for direct TA cloning and subsequent sequencing of unmodified PCR products since Taq DNA polymerase preferentially adds an adenosine (A) residue to the 3' end of the products under standard PCR conditions.
Ido
E
E
Laboratory of Pathogenic Viruses, Kyoto University, Japan. eido@virus.kyoto-u.ac.jp
Hayami
M
M
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction