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. 2013 Sep 23;41(Pt 5):1189–1194. doi: 10.1042/BST20130123

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© 2013 The author(s) has paid for this article to be freely available under the terms of the Creative Commons Attribution Licence (CC-BY)(http://creativecommons.org/licenses/by/3.0/) which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited.

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Green coloration indicates ideal performance, pale pink coloration indicates tolerable performance, and deep pink coloration indicates unacceptable performance, where non-degenerate methods (^a) include ‘small-intelligent libraries’ [8], Slonomics™ [10,11] and ProxiMAX. (A) Diversity was calculated using the formula d=1/(NΣ_kp_k²) [12] and is in agreement for a 12-mer peptide saturated with codon NNN [13]. (B) Ratios represent the theoretical relative concentrations of each individual gene combining any of the most common codons (leucine/arginine/serine, NNN/NNK; or leucine/valine, 22c trick) compared with each individual gene containing any combination of the rarest codons (^b) [methionine/tryptophan, NNN; cysteine/aspartate/glutamate/phenylalanine/histidine/isoleucine/lysine/methionine/asparagine/glutamine/tryptophan/tyrosine, NNK; or 18 codons (omitting leucine/valine), 22c trick]. (C) Truncation is calculated as the percentage of sequences that contain one or more termination codons within the saturated region. (D) NNN/NNK/trinucleotide oligonucleotides may be used as a DNA cassette, or as primers in PCR-based mutagenesis; Slonomics™ and ProxiMAX require a fixed number of oligonucleotides and the numbers of primers for the 22c trick [9] and small-intelligent libraries [8] were calculated using the formulae in the respective publications for saturating consecutive codons. (E) ^cAs dictated by degeneracy limitations.