i7® High-Fidelity DNA Polymerase 2x master mix is a ready to use premix which contains high-fidelity DNA Polymerase, dNTPs, MgCl2, PCR enhancers and stabilizers with optimized proprietary reaction buffer. i7® High-Fidelity DNA Polymerase 2x master mix has the high-fidelity, sensitivity and processivity with an error rate ~2.8×102 fold lower than Taq DNA polymerase, and significantly lower than the error rates of other proofreading enzymes.

Product Description

Intact Genomics (IG) i7® High-Fidelity DNA Polymerase 2x master mix is ready to use premix which contains i7 high-fidelity DNA Polymerase, dNTPs, MgCl2, PCR enhancers and stabilizers with optimized proprietary reaction buffer. i7 high fidelity DNA Polymerase is a genetically engineered, heat stable DNA polymerase which has 5´→3´ polymerase and 3´→5´ exonuclease (proofreading) activities. This enzyme has high fidelity, sensitivity and processivity with an error rate ~2.8×102-fold lower than Taq DNA polymerase, and significantly lower than other proofreading enzymes in the marketplace (1).

Activity Data
We have tested i7® high-fidelity 2x master mix with difficult templates for PCR amplification. Typical PCR results are shown below:

Applications

• Long and difficult template DNA amplification
• Cloning
• High-fidelity PCR
• Efficient for amplifying high GC content template DNA with magic enhancer.

 Product Includes

• i7^® high-fidelity DNA Polymerase 2x master mix

Storage Temperature
–20ºC

 Heat Inactivation
No

 Storage Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC3257 3259

Technical Support

Intact Genomics is committed to supporting the worldwide scientific research community by supplying the highest quality reagents. Each new lot of our products is tested to ensure they meet the quality standards and specifications designated for the product. Please follow the instructions carefully and contact us if additional assistance is needed. We appreciate your business and your feedback regarding the performance of our products in your applications.

Protocol

1. Thaw i7® high-fidelity 2x master mix, primer solutions, and mix thoroughly before use.
2. Prepare a reaction mix according to the following table:
(The reaction mix typically contains all the components
needed for PCR except the template DNA).

3. Mix the reaction mixture thoroughly.
4. Add template DNA to the individual PCR tube containing the reaction mixture.
5. Program the thermal cycler according to the manufacturer’s instructions. A typical PCR cycling program is outlined in the following table.

6. Place the PCR tubes in the thermal cycler and start the cycling program.

References

1. Frey, B. and Suppman, B. (1995). BioChemica. 2, 34-35.

Downloads

Manual – i7 High-Fidelity DNA Polymerase 2X Master Mix
MSDS – ig i7 High-Fidelity DNA Polymerase 2X MM