Directed evolution of Thal: Results
For random mutagenesis of thal adjustment of the mutation rate during epPCR is an essential precondition to obtain high-quality mutant libraries suitable for the directed enzyme evolution. A mutation frequency of 2-7 nucleotide mismatches per gene is recommended for the initial mutation approach. Regarding the 1.6 kb thal gene a mutation rate of 2 mutations/kb was desired and therefore epPCR optimized with regard to this target.
KeywordsRandom Mutagenesis Elution Fraction Crude Lysate Relative Conversion Preparative Synthesis
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