Multi-line split DNA synthesis: a novel combinatorial method to make high quality peptide libraries

• Main Authors: Ueno Shingo, Soramoto Sayaka, Tabuchi Ichiro, Husimi Yuzuru
• Format: Article
• Language: English
• Published: BMC 2004-09-01
• Series: BMC Biotechnology
• Online Access: http://www.biomedcentral.com/1472-6750/4/19

<p>Abstract</p> <p>Background</p> <p>We developed a method to make a various high quality random peptide libraries for evolutionary protein engineering based on a combinatorial DNA synthesis.</p> <p>Results</p> <p>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 10¹⁶diversity, and the construction of a library with random sequence coding 120 amino acids containing few stop codons.</p> <p>Conclusions</p> <p>Owing to the flexibility of the MLSDS method, it will be able to design various "rational" libraries by using bioinformatics databases.</p>