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. 2018 Aug 29;46(22):e132. doi: 10.1093/nar/gky769

Figure 2.

Figure 2.

Dominant cyclization of l-DNA using hairpins as internal promoters. (A) The solution structures of L643-4,24-4, L6416-4,37-4 and L643-4, determined by Mfold calculation under the conditions of [Mg2+] = 10 mM and 25°C. (B) Treatments of these l-DNAs with T4 DNA ligase. Lane 1, L643-4,24-4 without the T4 ligase treatment; lane 2, L643-4,24-4 treated with T4 DNA ligase in the presence of 12-nt splint which is complementary with the 6-nt sequences in the 3′- and 5′-ends of L643-4,24–4; lane 4, L6416-4,37-4 without the treatment; lane 5, L6416-4,37-4 treated with T4 DNA ligase in the presence of 12-nt splint. Lane 7, L643-4 without the treatment; lane 8, L643-4 treated with T4 DNA ligase in the presence of 12-nt splint. In lanes 3, 6 and 9, the products in lanes 2, 5 and 8 were further treated with Exonuclease I to remove non-cyclic products. The conditions for the T4 ligase reactions: [l-DNA]0 = 5 μM, [splint]0 = 10 μM and 10 U T4 DNA ligase in 1× T4 DNA ligase buffer at 25°C for 12 h.