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.

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 2.5x 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%.

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).

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
  • 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
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.