Hi Fidelity polymerase

Hi Fidelity polymerase is a mixture of thermostable enzymes. It is specifically developed to synthesize length of PCR product up to 25 kb and with low error rate. Hi Fi polymerase synthesizes higher yields of product from genomic DNA, cDNA, and bacterial cultures. It has 2.5 hours half life at 96oC and easily amplify PCR product of G-C rich or secondary structure DNA by adding G- C rich buffer.

-20°C

Buffer containing 25mM MgCl₂

One unit is defined as the amount of enzyme that will incorporate 10nmole of dNTP into acid-insoluble material in 30 minutes at 74ºC. The reaction conditions are: 50mM Tris-HCl pH8.8, 50mM NaCl, 5mM MgCl₂, 200uM each of dATP, dCTP, dGTP, dTTP, 10 mg activated calf thymus DNA and 0.1mg/ml BSA in a final volume of 50μl.

Hi Fidelity Polymerase is suitable for amplifying targets up to 25 kb from the following templates:

Amplification of longer targets (up to 15 kb) may be possible, but may require more template and longer elongation times.

Use 0.3 mM per primer as a general starting point. For larger amounts of template (e.g., 200 ng genomic DNA), increasing the concentration up to 0.5 μM per primer may improve yield.

The annealing temperature is slightly higher than with Typical PCR. The optimal annealing temperature should be ~2°C lower than the Tm of the primers used. A range of 58–68°C is recommended.

As little as 30 seconds per kb is suitable for most targets. Use up to 60 seconds per kb for maximum yield.

IMPORTANT: Annealing temperature should be 2-6ºC lower than the primer melting temperature.

Shipping and temporary storage at -20 and for up to 1 month at room temperature has no detrimental effects on the quality of Hi Fidelity DNA polymerase.