Recently, our team successfully delivered a highly customized variant library featuring 16 variable positions, including predefined amino acid biases (e.g., Thr at X2 and Ser at X3 enriched at a 3:1 ratio over other amino acids). The project also required a 109-scale library, >50% correct sequence rate, complete BbsI restriction site removal, and E. coli codon optimization.
 
Using our trinucleotide-mediated (trimer) oligo synthesis, we precisely controlled the amino acid composition at every variable position. NGS-based quality analysis confirmed outstanding performance:
  • 109 library diversity achieved, with 50.01% valid sequences, meeting the project's >50% accuracy requirement.
  • Accurate amino acid bias control, with Thr (X2) and Ser (X3) frequenciesmatching the designed 3:1 ratio, significantly reducing distribution bias compared with conventional degenerate codons.
  • Excellent positional coverage, with measured amino acid frequencies across all variable sites closely matching the theoretical design, ensuring balanced diversity throughout the library.
This project demonstrates the strength of Trimer technology for constructing highly complex variant libraries requiring precise amino acid distributions and exceptional sequence quality.
 
If you're planning a similar library or would like to review the complete NGS analysis, we'd be happy to discuss your project and recommend the most suitable library design strategy.

 
 
 
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Synbio Technologies LLC
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