We have constructed a short gene with a single step assembly PCR without any additional amplification primers with Taq and Pfu polymerases. Since the Taq polymerase is a common and conventional enzyme for PCR reactions, we have analyzed various effects on its efficiency. Eventually, we have been able to synthesize the gene in less than 40 minutes by Taq polymerase.
single step assembly PCR; efficiency; amplification primer; polymerase