Fig. 2.

Optimized multi-site NM protocol yields near-comprehensive coverage of combinatorial libraries. Results from the optimized ‘dual directions’ strategy for generating comprehensive combinatorial libraries. (left) Histograms for frequency of the total number of variants and (right) violin plots of the frequency of each variant relative to the number of mutations from CR6261 (replicate 1 A; replicate 2 B) or CR9114 (replicate 1 C; replicate 2 D). For all panels, the violin plots were normalized where a log₁₀ frequency of zero corresponds to a mutation not being observed by deep sequencing. The dashed gray lines indicate the relative frequency for a uniformly distributed library.