The TA cloning method takes advantage of the terminal transferase activity of some DNA polymerases such as Taq polymerase. This enzyme adds a single, 3'-A overhang to each end of the PCR product. This makes it possible to clone this PCR product directly into a linearized cloning vector with single, 3'-T overhangs. To clone such PCR product the linearized cloning vector with single, 3'-T overhangs called a T- vector are used. The PCR products with dA overhang, are mixed with this vector in high proportion. The complementary overhangs of "T" vector and PCR product will get hybridized and ultimate joining of two DNA fragments accomplished by T4 DNA ligase.
TA Cloning with SimVector
SimVector automatically generates graphical DNA sequence maps of designed recombinant DNA molecules displaying vector - donor relationships and the corresponding cloning protocol. Use commercial T-vector or design your own T-vector using restriction digestion and recombine with the donor in sense or anti-sense orientation to generate highly accurate recombinant DNA sequence map.