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I7® High-Fidelity DNA Polymerase
i7® High-Fidelity DNA Polymerase 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.
Price range: $152.00 through $335.00
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Product Description
Intact Genomics (IG) 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 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 in the marketplace (1). It is ideal for cloning and can be used for long (up to 20kb) or difficult amplicons. This product is supplied with the Intact Genomics proprietary 2.5x i7® PCR reaction buffer containing MgCl2 with a final (1x) concentration of 2 mM. This buffer allows for amplification of non GC rich templates and of GC rich templates up to 84%.
Protein Purity
The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).
Product Source
E. coli strain expressing genetically engineered i7® High-Fidelity DNA Polymerase gene.
Comparison Data
We have tested this DNA Polymerase activity with λ DNA and other difficult templates for PCR amplification up to 20kb. Intact Genomics (IG) i7® high-fidelity DNA Polymerase generates robust and high-quality PCR products in comparison with other high-fidelity DNA polymerases available in the marketplace (data 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
- 2.5x i7® PCR Buffer with Mg2+
Storage Buffer
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC
Storage Temperature
–20ºC
Heat Inactivation
No
Quality Control Assays
Free from detectable nuclease activities.
Unit Definition
One unit is defined as the amount of enzyme that
incorporates 10 nmoles of dNTP into acid-insoluble form in 30 minutes at 72º C.
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Protocol
1. Thaw 2.5x i7 PCR Buffer, dNTP 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.
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