Diversify PCR Random Mutagenesis Kit for sequence mutations

The Diversify PCR Random Mutagenesis Kit employs a nontoxic method for mutagenizing sequences of up to 4.0 kb in length. The kit broadly distributes all possible mutations without creating hot spots. By allowing you to manipulate mutagenic conditions, the Diversify PCR Random Mutagenesis Kit provides optimal mutagenesis of sequences over an exceptionally broad size range. Select mutation rates from two to eight mutations per 1,000 bp simply by varying the amounts of two key reagents—manganese (Mn2+) and dGTP (Cadwell and Joyce 1992; Leung, Chen, and Goeddel 1989).

The Diversify PCR Random Mutagenesis Kit contains our Titanium Taq PCR system to reliably amplify DNA fragments of up to 4.0 kb in length. This extended amplification range makes the Diversify kit ideal for mutating operons, plasmids, and sequences corresponding to large proteins, in addition to short DNA fragments. The kit also contains a rapid control reaction that allows the relative comparison of mutation rates in the control fragment in just two hours following PCR. In addition to the Titanium Taq polymerase mix, the Diversify PCR Random Mutagenesis Kit includes all of the reagents and buffers needed for mutagenesis.

Overview

Controlled, random mutagenesis for investigating protein function
Precise control of mutation rate
No mutational hot spots
Obtain all possible base substitutions

Applications

Random mutagenesis

Mutation rates can be controlled by buffer conditions

Mutation rates can be controlled by buffer conditions. Mutation rates for buffer conditions 1, 5, and 9 were obtained from extensive DNA sequencing (>15,000 bp each). Remaining mutation rates were standardized to sequencing data using an in vivo mutagenesis assay (3).

The Diversify PCR Random Mutagenesis Kit amplifies fragments as large as 4.0 kb

The Diversify PCR Random Mutagenesis Kit amplifies fragments as large as 4.0 kb. A 4.0-kb plasmid was amplified under the buffer conditions shown in Lanes 1–9 correspond to PCR buffer conditions 1–9. Lane M: DNA size marker.

References

Cadwell, R. C. & Joyce, G. F. Randomization of genes by PCR mutagenesis. PCR Methods Appl. 2, 28–33 (1992).

Leung, D., Chen, E., and Goeddel, D. V. PCR-based random mutagenesis using skewed nucleotide concentrations. Technique 1, 1–15 (1989).

Product citations

Mo, J. Y., Maki, H. & Sekiguchi, M. Mutational specificity of the dnaE173 mutator associated with a defect in the catalytic subunit of DNA polymerase III of Escherichia coli. J. Mol. Biol. 222, 925–36 (1991).