Different amounts of beads and xyloglucan were used in the different rounds of selection.
Many sequence classes peaked in frequency at different rounds of selection, but in this case, one class appeared to consistently maintain a high frequency.
In the regular panning method, small differences were in the stringency of selection between different rounds of panning, but in modified subtractive panning after two or three rounds of selection, the antigen-binding clones showed a clear enrichment.
In addition, we used growth in the presence of ethanol as a selection method between different rounds of genome shuffling in some approaches.
For both genome shuffling based on random mating and on targeted mating, we also investigated the dynamics of not applying any selection in between the different rounds of genome shuffling.
Third, we also performed the different rounds of random genome shuffling in the absence of any selection.
Figure 2 CHs are reallocated in different rounds of transmission.
To compare ethanol tolerance between different rounds of shuffling, for each replicate and the unselected F1 pool 24, random isolates were obtained after streaking frozen populations, which were obtained after washing off the biomass after selection, on YPD agar.
In the third round of selection, two different strategies, which differed in the number of washes and the concentration of the coating conjugates, were used to select for specific binders to monensin.
Because the library is compact and a protease sensitive helper phage, KM13 [ 37] was used during bio-panning, only one or two round of selection were performed on eight different antigens.
