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Randomize Gene Sequences with New PCR Mutagenesis Kit



GeneMorph PCR mutagenesis kit produces a unique mutational spectrum

Janice Cline Holly Hogrefe

Stratagene

The GeneMorph PCR mutagenesis kit simplifies the process of constructing mutant libraries for protein evolution and structure-function studies. Errors are introduced during PCR at low, moderate, or high mutation frequencies using a single set of optimal reaction conditions. The kit features Mutazyme DNA polymerase, a novel error-prone DNA polymerase mutant developed at Stratagene. Mutazyme DNA polymerase produces all possible transition and transversion mutations with minimal bias, and its mutational spectrum is distinct from that of Taq DNA polymerase. Unlike current Taq- based protocols, the GeneMorph method is robust and can be applied to mutagenesis of long targets (1-10 kb).

Error-prone PCR is a random mutagenesis technique for introducing amino acid changes into proteins. Mutations are deliberately introduced during PCR through the use of error-prone DNA polymerases and reaction conditions.1,2 Randomized DNA sequences are cloned into expression vectors, and the resulting mutant libraries are screened for altered or improved protein activity. To analyze structure-function relationships, mutation frequencies of one amino acid change (1-2 nucleotide changes) per gene are desired, while directed evolution studies typically empl
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