Extended Data Figure 5.

Analysis of the regiospecificity of ligation. a, D-RNA substrates and template for ligation, catalyzed by the L-16.12tx enzyme. Dot indicates the ligation junction, which is also the site for RNase A cleavage that is closest to the 3′ end of the ligated product. The downstream substrate is labeled at the 3′ end with fluorescein (circled F). b, RNase A digestion of the ligated products (LP) in comparison to authentic all-3′,5′-linked RNA of the same sequence (S10). Reaction conditions: 0–100 μg/μL RNase A, 2 μM RNA, 50 mM Tris (pH 7.6), 23 °C, 1 min.