Figure 4.
Sequence analysis of products of nonenzymatic primer-extension. (a) Top: Primer-extension was carried out on 2.0 μM template T9 (AGAGAG) in the presence of 40 mM 2-MeImpC and 50 mM 2-MeImpU* (U* = U, s2U, or s2T) on ice for 7 days. Bottom: Products were sequenced, and the sequence reads binned according to the number of nucleotides added to the primer; Y = C or U*. Reaction conditions: 2.0 μM primer P2, 200 mM HEPES pH 7.0, 100 mM MgCl2. (b) Top: Primer-extension was carried out on 2.0 μM template T5 (UC5), T6 (s2UC5), or T7 (s2TC5) in the presence of 40 mM 2-MeImpG and 50 mM 2-MeImpA on ice for 7 days. Bottom: Products were sequenced and the sequence reads binned according to the number of nucleotides added to the primer; R = A or G.