Precision Synthesis of Variant Libraries Enables Comprehensive Interrogation of Single-Site Variant Space

Single Site Variant Libraries (SSVL) enable the interrogation of sequence space to identify key residues in protein structure and function. Traditional methods of generating variant diversity, such as error- prone PCR and site-directed mutagenesis using degenerate or semi-degenerate oligonucleotides, generate only a random sampling of mutations in the gene sequence. These methods can create premature stop codons and incorporate redundant codons, which leads to poor control and uneven distribution of diversity. Twist Bioscience’s silicon-based DNA synthesis platform, on the other hand, enables precise synthesis of variants to allow unprecedented, efficient, and comprehensive interrogation of the single variant space.