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.