Similar to error-prone PCR-based random mutagenesis, AquaMutant may be used to introduce random mutations limited to defined DNA sequence (e.g., a gene of interest) by treating a corresponding PCR product or restriction fragment in vitro. The treated DNA fragment, with or without further PCR amplification, may subsequently be ligated into a plasmid vector for expression.
However, unlike error-prone PCR-based random mutagenesis, AquaMutant also may be used to introduce random mutations in live cells or model organisms. Such in vivo random mutagenesis has many advantages: it is simple to carry out; it does not require prior genetic knowledge to study a phenotype; it can mutate multiple related genes concurrently and overcome the limitation of in vitro random mutagenesis where the mutation or knockout of a single gene may be compensated by other genes; and it is particularly suited for direct evolution to create mutants with desired properties by multiple rounds of mutation-selection.
Mutant libraries created by usage of AquaMutant are useful tools in life science research to investigate biological processes and disease mechanisms, screen for enzymes and antibodies with improved properties, and use directed evolution to generate mutants with desired functionalities and phenotypes.
AquaMutant is intended for in vitro laboratory research only.