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.
i7® High-Fidelity DNA Polymerase
$148.00 – $325.00
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). i7® high-fidelity DNA polymerase 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%.
The physical purity of this enzyme is ≥98% as assessed by SDS-PAGE with Coomassie® blue staining (see figure below).
E. coli strain expressing genetically engineered i7® High-Fidelity DNA Polymerase gene.
We have tested i7® High-Fidelity 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):
- Long and difficult template DNA amplification
- High-fidelity PCR
- Efficient for amplifying high GC content template DNA with magic enhancer
- i7® High-Fidelity DNA Polymerase
- 2.5x i7® PCR Buffer with Mg2+
50 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.5 @ 25ºC
Quality Control Assays
i7® High-Fidelity DNA Polymerase is free from detectable nuclease activities.
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.
200 Units, 500 Units
- Thaw 2.5x i7 PCR Buffer, dNTP mix, primer solutions and mix thoroughly before use.
- Prepare a reaction mix according to the following table: (The reaction mix typically contains all the components needed for PCR except the template DNA. )
- Mix the reaction mixture thoroughly.
- Add template DNA to the individual PCR tube containing the reaction mixture.
- Program the thermal cycler according to the manufacturer’s instructions. A typical PCR cycling program is outlined in the following table.
- Place the PCR tubes in the thermal cycler and start the cycling program.