Figure 2. Optimization of translation efficiency and confirmation of mRNA/cDNA-peptide conjugates. Gel images of the mRNA-peptide conjugate formation under various translation conditions. (A) Difference in length of translation before salt addition. Number indicates different incubation time (in minutes) carried out at 37 degrees C. (B) Difference in salt concentrations with original concentration (1x) as 750 mM KCl, 65 mM MgCl2 used in the previous studies (Naimuddin et al., 2011; Naimuddin & Kubo, 2016) (C) Difference in lengths of incubation time (in minutes) after salt addition. (D) Trypsin digestion of mRNA-tag and mRNA-peptide conjugate products. (E) Detection of FITC fluorescence signal from the mRNA-tag, mRNA-peptide conjugate, and mRNA/cDNA-peptide conjugate products (left) and detection of FLAG epitope sequence using chemiluminescent Western blotting (right) from the same membrane.