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i DNA Evolution

i DNA Evolution Our proprietary i DNA Evolution Technology We developed a simple and economical method for preparing a mutant protein library useful for the directed evolution of a protein, and have achieved a transposon-mediated Random Codon based Mutagenesis (RCM) method. The error-prone PCR method is widely used for constructing a mutant DNA library of a gene. In this method, a mutant library is prepared by controlling the polymerization conditions to change the error rate of a polymerase. However, the error-prone PCR method has a problem in that it is difficult to control the error rate appropriately to obtain a desired frequency of mutation. Moreover, the frequency of co-occurrence of more than one base substitution within a codon is too low, so that the number of mutant amino acids for a given amino acid residue is limited. If you use our RCM method, you will prepare a library of mutant polynucleotides with high diversity through transposon-mediated random substitution, insertion or deletion of nucleotides on a polynucleotide coding for a target protein. (Patent KR 0446417, US 6955879, JP 3777158) Next, we developed a new approach to in vitro DNA recombination technique termed Recombined Extension on Truncated Templates(RETT). This method does not use DNA endonucleases for generation of shuffling blocks. Instead, it makes unidirectional single-stranded DNA (ssDNA) fragments by either DNA polymerase in the presence of random primers or serial deletion with exonucle

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