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. 2019 Jun 10;141(26):10481–10488. doi: 10.1021/jacs.9b04858

Figure 2.

Figure 2

Chemical copying of RNA template 3′-GCCC into 3′-NP-DNA using (A) 3′-NH2-2AIpddC alone, (B) 3′-NH2-2AIpddC and 3′-NH2-5′-PO3-ddG, and (C) 3′-NH2-2AIpddC and 3′-NH2-2AIpddG. The nucleotide highlighted in red indicates the 3′-NH2-ddC at the end of the primer. All primer extension reactions were conducted at pH 8.5, 25 °C, and 200 mM Na+-bicine with 10 mM of each mononucleotide and 50 mM Mg2+. Reaction products were analyzed by polyacrylamide gel electrophoresis (PAGE). The RNA templating region is indicated in blue. The slow primer extension observed in the presence of 3′-NH2-2AIpddC alone (t1/2 = 16 ± 1 min), or 3′-NH2-2AIpddC plus 3′-NH2-ddG-5′-monophosphate (t1/2 = 26 ± 2 min) relative to 3′-NH2-2AIpddC plus 3′-NH2-2AIpddG (t1/2 < 0.5 min) suggests that the polymerization reaction proceeds via the imidazolium-bridged dinucleotide intermediate C-2AI-G. P, primer. Cyt, cytosine. Gua, guanine.