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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 1.

Figure 1

Formats for in vitro evolution of RNA ligation activity. (A) Substrate-A format, employing a separate 5′-biotinylated molecule bearing a 3′-hydroxyl. The primer for reverse transcription is pre-hybridized to the 3′ end of the enzyme to prevent interaction of this region with the substrate. (B) Substrate-B format, employing a separate 3′-biotinylated molecule bearing a 5′-triphosphate. Curved arrow indicates the site of ligation. Circled “B” indicates the biotin moiety. Open lines within the enzyme indicate nucleotides that were mutagenized at 21% degeneracy in the starting population.