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GeneMorph II random mutagenesis kit
上海恒斐生物科技有限公司
-20
30 reactions
Random mutagenesis is a powerful tool for elucidating protein structure-function relationships and for modifying proteins to improve or alter their characteristics. Error prone PCR is a random mutagenesis technique for generating amino acid substitutions in proteins by introducing mutations into a gene during PCR. Mutations are deliberately introduced through the use of error prone DNA polymerases and/or reaction conditions. The mutated PCR products are then cloned into an expression vector and the resulting mutant library can be screened for changes in protein activity. Random mutagenesis allows researchers to identify beneficial mutations in the absence of structural information, or when such mutations are difficult to predict from protein structure.
The mutational bias exhibited by error prone PCR enzymes undoubtedly skews representation of random mutant libraries, diminishing the effective size of the collection produced by error prone PCR. Mutazyme II DNA polymerase is a novel error prone PCR enzyme blend, formulated to provide useful mutation rates with minimal mutational bias. Mutazyme II is a blend of two error prone DNA polymerases Mutazyme I DNA polymerase (from the original GeneMorph Random Mutagenesis Kit) and a novel Taq DNA polymerase mutant that exhibits increased misinsertion and misextension frequencies compared to wild type Taq. For the Mutazyme II polymerase formulation, the Mutazyme I polymerase and the Taq polymerase mutant have been combined to produce a less biased mutational spectrum with equivalent mutation rates at A’s and T’s vs. G’s and C’s (see Figure 1). Therefore, libraries created with Mutazyme II should exhibit greater mutant representation compared to libraries generated with other enzymes. However, the original GeneMorph I kit favors mutations at G’s and C’s which in some cases may be desirable.
With the GeneMorph II random mutagenesis kit, mutation rates of 1–16 mutations per kb can be achieved using a single set of buffer conditions (MgCl2, balanced dNTPs) optimized for high product yield. The desired mutation rate can be controlled simply by varying the initial amount of target DNA in the reaction or the number of amplification cycles performed
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