New England Biolabs, M0294S, Tth Endonuclease IV

Related Categories DNA Repair Enzymes and Structure-specific Endonucleases Specification Unit Definition One unit is defined as the amount of enzyme required to cleave 1 pmol of a 60-mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 μl in 1 hour at 65°C. * An AP site is created by treating 10 pmol of a 60-mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C. Reaction Conditions 1X ThermoPol® Reaction Buffer Incubate at 65°C 1X ThermoPol® Reaction Buffer 20 mM Tris-HCl 10 mM (NH4)2SO4 10 mM KCl 2 mM MgSO4 0.1% Triton® X-100 (pH 8.8 @ 25°C) Usage Concentration 10,000 units/ml Storage Buffer 10 mM Tris-HCl 100 mM KCl 1 mM DTT 0.1 mM EDTA 50% Glycerol 0.1% Triton® X-100 pH 7.4 @ 25°C Heat Inactivation No Unit Assay Conditions 1X ThermoPol Reaction Buffer containing 5 pmol of fluorescently labeled oligonucleotide duplex in a total reaction volume of 10 μl. FAQ Q: How does Tth Endonuclease IV perform in PCR? A: Tth EndoIV can survive typical PCR cycling. The percentage surviving will depend on the specific conditions. It has a significantly higher thermostability in ThermoPol buffer than Standard Taq buffer and we recommend adding Zn++ (usually ZnCl2) to the reaction to 25 micromolar to ensure the enzymes thermostability. We have not seen this level of Zn++ to effect PCR reactions.