Figure 3.

Effects of the concentration of T4 ligase buffer on the efficiencies of cyclization of l-DNA₆₆ and its polymerization. All the DNA substrate was added to the mixture all at once at the beginning of the reaction. The selectivity for the formation of c-DNA₆₆ is presented below the corresponding band. The reaction conditions: [l-DNA₆₆]₀ = 1 μM; [splint-12 nt]₀ = 2 μM; 5 U T4 DNA ligase at 20°C and 12 h. Note that 1 × T4 ligase buffer contains 10 mM MgCl₂, 500 μM ATP, 10 mM DTT and 40 mM Tris-HCl.