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. Author manuscript; available in PMC: 2015 Feb 20.
Published in final edited form as: Chem Biol. 2014 Jan 2;21(2):238–245. doi: 10.1016/j.chembiol.2013.12.004

Figure 2.

Figure 2

Assay of ligation activity for successive populations of enzymes over the course of in vitro evolution. Reactions were performed in the substrate-A format for the E1 enzyme (wt), starting population (0), and populations obtained after rounds 1–10, employing 250 (black), 100 (gray), or 50 (white) nM substrate, always in two-fold excess over enzyme. Reaction conditions: 25 mM MgCl2, pH 8.5, 42 °C.