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Identification of hot spot mutations through amplicon deep sequencing. A, schematic of mutation frequency at each amino acid in the adiponectin globular domain (residues 109–244) after the final FACS2 selection round. B, residues showing more than a 10% mutation rate in FACS round 2 were then analyzed to identify preferred substitutions. C, hot spot mutations are represented as side chains in a ribbon representation of the structure.
