Tfl DNA Polymerase is a thermostable enzyme of approximately 94kDa isolated from Thermus flavus. The enzyme replicates DNA at 74°C and exhibits a half-life of 40 minutes at 95°C. Tfl DNA Polymerase catalyzes the polymerization of nucleotides into duplex DNA in the 5´→3´ direction in the presence of magnesium and the polymerization of nucleotides into DNA using an RNA template in the 5´→3´ direction in the presence of manganese. The enzyme also possesses a 5´→3´ exonuclease activity. Tfl DNA Polymerase is recommended for use in PCR and primer extension reactions at elevated temperatures.
Tfl DNA Polymerase is provided with a 10X Reaction Buffer that does not contain magnesium. Sufficient 25mM MgSO4 is provided separately to allow optimization of enzyme performance under different conditions.
- 10X Reaction Buffer: 200mM Tris-acetate (pH 8.9 at 25°C), 100mM ammonium sulfate.
- Unit Definition: One unit is defined as the amount of enzyme that will incorporate 10nmol of dNTPs into TCA-insoluble material in 30 minutes at 74°C.
- Source: Thermus flavus.
- Storage Buffer: 20mM Tris-HCl (pH 8.0 at 25°C), 100mM KCl, 0.1mM EDTA, 1mM DTT, 0.5% Nonidet®-P40, 0.5% Tween® 20 and 50% glycerol.
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- Kaledin, A.S. et al. (1981) Biokhimiia 46, 1576–84.