Temperature and concentration effect. 20% polyacrylamide denaturing gel is stained with SybrGold® gel stain for 20 min. Line M is a marker line. (A) Effect of the temperature on polycondensation efficiency of nTGp with nCAp in 1:1 ratio. Total nucleotide concentration is 20 mM. (B) Concentration effect on nCGp polymerization. Reaction products are visualized after 48 h of reaction. (C) Comparison of the reaction rates at different concentrations of the dinucleotide. 5 mM nCGp and 50 mM nCGp show similar reaction rates and size of the final product.