3.1. Description of the Transfer-PCR (TPCR) process
3.2. Application of TPCR for DNA cloning
3.3. Application of TPCR for multiple-site targeted mutagenesis
3. Results
Fig.1. Schematic presentation of the Transfer-PCR (TPCR) process. (A) TPCR for DNA cloning. The target gene for cloning is marked in blue. The donor and the recipient
plasmids are marked in green and brown, respectively. Primers are indicated by arrows that contain a gene-specific sequences at the 30 end (solid blue line) and a vectorspecific
sequences corresponding to the integration sites on the recipient vector, at the 50 end (solid brown line). Dashed blue line indicates formation of an amplification
product. (B) TPCR for protein engineering. The target gene for mutagenesis is marked in blue. Blue arrows indicate mutagenic primers with the mutation sites shown by red
stars. For both (A) and (B), red X over the plasmid indicates elimination of the wild-type (donor or parental) plasmid by DpnI treatment.