Figure 2

Effect of EDTA treatment on the capturing of mRNA-protein fusions by SA-beads. mRNA-protein fusion molecules in the translation reaction mixture with (+) or without (−) EDTA were incubated with SA-beads [the ratio of the biotin-binding capacity of the SA-beads to the total amount of puromycin-linker conjugate is 200 to 1]. (A) The remaining mRNA-protein fusion molecules in each translation reaction mixture were analyzed by 4% stacking–6% separating SDS PAGE containing 8 M urea. The input mRNA-linker conjugates and translation reaction mixture are shown in lane I and lane T, respectively. (B) Binding efficiencies are calculated by: [band intensity of the mRNA-protein fusion molecules in lane T] - [band intensity of the remaining mRNA-protein fusion molecules in each lane indicated by “+” or “–” EDTA].