i7™ Hot Start 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™ Hot Start High-Fidelity DNA Polymerase
$199.00 – $399.00
Description
Intact Genomics (IG) i7 Hot Start High-Fidelity DNA Polymerase is a genetically engineered, heat stable DNA polymerase which has 5´→3´ polymerase and 3´→5´ exonuclease (proofreading) activities. i7 Hot Start High-Fidelity DNA Polymerase is chemically modified that leads to complete inactivation of the polymerase until the initial heat activation step at the start of PCR. Hot start PCR reduces non-specific amplification during setup stages of the reaction and helps increase PCR specificity and sensitivity. 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 Hot Start 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 5x PCR reaction buffer containing MgCl2 with a final (1x) concentration of 2 mM, and 5x magic enhancer that enables efficient amplification of GC rich templates up to 84%.
Activity data
i7 Hot Start High-Fidelity DNA Polymerase generates robust and high-quality PCR products with difficult templates (Fig. A). PCR extension temperatures can be used between 68 to 72º C (Fig. B). This enzyme is resistant to different PCR inhibitors such as heparin, humic acid and xylan (Fig. C).
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 Hot Start High-Fidelity DNA Polymerase
- 5x i7 PCR Buffer with Mg2+
- 5x Magic Enhancer
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
i7 Hot Start High-Fidelity DNA Polymerase is 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.
3281 3283
Additional information
| Unit | 200 Units, 500 Units |
|---|
Protocol
- Thaw i7 PCR Buffer, dNTP mix, primer solutions, 5x magic enhancer (if required) 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.
References
Frey, B. and Suppman, B. (1995). BioChemica. 2, 34-35.
