Fig. 2.

Dynamic ¹⁹F NMR spectra (−116 ~ −121 ppm) of various FAF-G:C match template/primer models at n−1, n, n+3, and n+6 positions (n=primer terminus; G=FAF-adduct). Spectra of all sequences obtained at seven standard temperatures (labeled as 5, 10, 20, 30, 40, 50, and 60 °C). Additional temperatures were recorded for n−1 (23, 25 °C), n (12, 15, 21, 22, 23, 25 °C), n+1 (15, 22, 25 °C), n+3 (15, 18 °C), and n+6 (15, 25, 33, 35 °C). Inset shows NOESY/exchange spectra of the n+6 dC-match duplex at 5 and 20 °C (dotted red circles indicate the areas for possible off-diagonal cross peaks). Tm (10⁻⁴ M) of the n+3 template-primer is 24 ~ 29 ºC. Tm (10⁻⁴) of the n+6 dC template-primer is 45.6±0.4 ºC