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. 2014 Sep 10;42(18):11752–11762. doi: 10.1093/nar/gku740

Figure 7.

Figure 7.

(Left) Cleavage assay reactions (A,B,C) of designed hammerhead (wild-type), mutant C116G and mutant G142U. For the wild-type (A), mutant C116G (B), and mutant G142U (C) gel images, lane 1 is the undigested RNA (full-length, FT), lanes 2–5 are reactions in cleavage buffer (50 mM Tris pH 7.5, 5 mM MgCl2) at the 0 s, 30 min, 5 h and 24 h time points respectively (5′ and 3′ cleavage products indicated). For the wild-type (A), lane 6 is a reaction lacking Mg (50 mM tris pH 7.5) incubated for 24 h. It is evident that cleavage only occurs for the wild-type sequence, and when Mg is present. (Right) Cleavage time series curve (D) for the 166 nt designed hammerhead, with observed cleavage rate of 1.3/min with an Fmax of 0.47 and MSE of 0.0026. This construct displays kinetics comparable with that of wild-type hammerheads, although the cleavage amount Fmax is much lower than that of wild-type hammerheads.