Journal
BMC BIOTECHNOLOGY
Volume 4, Issue -, Pages -Publisher
BMC
DOI: 10.1186/1472-6750-4-19
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Background: We developed a method to make a various high quality random peptide libraries for evolutionary protein engineering based on a combinatorial DNA synthesis. Results: A split synthesis in codon units was performed with mixtures of bases optimally designed by using a Genetic Algorithm program. It required only standard DNA synthetic reagents and standard DNA synthesizers in three lines. This multi-line split DNA synthesis (MLSDS) is simply realized by adding a mix-and-split process to normal DNA synthesis protocol. Superiority of MLSDS method over other methods was shown. We demonstrated the synthesis of oligonucleotide libraries with 1016 diversity, and the construction of a library with random sequence coding 120 amino acids containing few stop codons. Conclusions: Owing to the flexibility of the MLSDS method, it will be able to design various rational libraries by using bioinformatics databases.
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