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Diversify® PCR Random Mutagenesis Kit

 
 

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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.

The Diversify Method

By allowing you to manipulate mutagenesis conditions, 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 (Figure 1; 1, 2).

The Diversify method utilizes Clontech's 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.

The Diversify PCR Random Mutagenesis Kit includes the TITANIUM Taq PCR System and all mutagenesis buffers and reagents.

 
 
 
 

Figure 1. 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).

 
 

Figure 2. 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 Figure 1. Lanes 1–9 correspond to PCR buffer conditions 1–9. Lane M: DNA size marker.

 
 
 
 

Applications

  • Create all possible base mutations at a random level

    • References

    1. Leung, D. W., et al. (1989) Technique 1:11–15.
    2. Cadwell, R. C. & Joyce, G. F. (1992) PCR Methods Appl. 2(1):28–33.
    3. Mo, J.-Y., et al. (1991) J. Mol. Biol. 222(4):925–936.

    Components

    TITANIUM Taq DNA Polymerase
    TITANIUM Taq PCR Buffer
    Diversify dNTP the Mix
    dGTP
    dNTP Mix
    Manganese Sulfate
    Purified H2O
    Control PCR Template & Primer Mix
    Taq I Restriction Enzyme

    Storage Conditions

    –20°C
     
     
     
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    Diversify PCR Random Mutagenesis Kit 30 rxns 630703 $602.00 USD    Add to cart
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