New England Biolabs, M0261L, Therminator™ DNA Polymerase

Therminator DNA Polymerase is a 9°N™ DNA Polymerase variant with an enhanced ability to incorporate modified substrates such as dideoxynucleotides, ribonucleotides and acyclonucleotides (1,2). Related Categories DNA Labeling Specification Unit Definition One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 75°C. Reaction Conditions 1X ThermoPol® Reaction Buffer 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) Storage Buffer 10 mM Tris-HCl 100 mM KCl 1 mM DTT 0.1 mM EDTA 50% Glycerol pH 7.4 @ 25°C Heat Inactivation No Molecular Weight Theoretical: 90000 daltons 5' - 3' Exonuclease No 3' - 5' Exonuclease No Strand Displacement + Unit Assay Conditions 1X ThermoPol Reaction Buffer, 200 µM dNTPs including [ 3 H]-dTTP and 15 nM primed single-stranded M13mp18. FAQ Q: Can Therminator™ DNA Polymerase be used in other buffers? A: Substituting buffers is not recommended. Q: When should Therminator™ DNA Polymerase be used? A: When modified nucleotides such as dideoxynucleotides, ribonucleotides, and acyclonucleotides must be incorporated. Q: Can Therminator™ DNA Polymerase be substituted for ThermoSequenase™? A: Yes, both incorporate modified nucleotides. For labs requiring large amounts of polymerase, using Therminator DNA Polymerase could result in considerable cost savings. Optimizing the Therminator DNA Polymerase reaction for various applications may be required. Q: Can Therminator™ DNA Polymerase be used for sequencing? A: Therminator DNA Polymerase can be used for sequencing but optimization and licensing may be required. Using the standard ABI nucleotide mix gives less than optimal results. Q: Why are some of the A's in my sequence weak when using Therminator™ DNA Polymerase? A: Under the sequencing conditions tested, A after C consistently gives a weak signal.