Figure 2

Pilot experiment demonstrating the cloning specificity of the TS-HR technique. (A) Schematic representation of the homologous regions between the vectors and the DNA fragments. (B) Result of the conventional homologous recombination (left) and the TS-HR method (right). The PCR-amplified V gene sequence and mock DNA were mixed in the indicated molar ratios and introduced into E. coli with either non-selective (pNS-hIgG) or selective vector (pTS-hIgG). Colony PCR was performed, and the amplified DNA fragments were separated by agarose gel electrophoresis. Forty colonies were analyzed, and data from a representative experiment are presented; the cloning specificity is expressed as a percentage of the total number of colonies analyzed.