Figure 5.

Assembly of long oligonucleotides in solution and inside vesicles by tetramer ligation. a) The splint template is non‐activated tetramer 5′‐AGUC‐3′. The ligator is 5′‐MeImp‐CUGA${}_{\mathrm{NH}{}_2}$ . Cy3‐labeled primer with a GA overhang is used to visualize the reaction. b) Scheme of the reaction inside vesicles. c) Representative gels are shown for reactions in solution and inside DA/DOH/GDM vesicles. Reactions in solution contained 4 μm primer, 6 μm template, 0.5 mm AGUC, 0.5 mm CUGA${}_{\mathrm{NH}{}_2}$ , 200 mm HEPES, pH 8.0, 0 or 100 mm MgCl₂, 0 or 1 mm EB, 0 or 100 mm HEI, at room temperature. Reactions in vesicles were performed with 200 mm Bicine, pH 8.5, and the addition of 0.5 mm AGUC, 0.5 mm CUGA${}_{\mathrm{NH}{}_2}$ , 50 mm MgCl₂, 200 mm sodium citrate, 0 or 1 mm EB outside the vesicle.