Figure 5. Translation of phi29 DNA polymerase from newly synthesized DNA in the TTcDR reaction.

A) Experimental procedure. First, we performed the optimized TTcDR reaction without [³⁵S]-methionine in the presence or absence of dNTPs, and one-tenth of the mixture was transferred to the second reaction mixture, which contained [³⁵S]-methionine, to detect translation from the replicated DNA product in the first reaction. After incubation at 30 °C for 12 h, an aliquot was used for 10% SDS-PAGE and autoradiography. B) Translation results. Increased translation of the DNA polymerase was detected when the first reaction contained dNTPs, indicating that the translation occurred from the DNA produced in the first reaction.