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EconoTaq DNA Polymerase is provided with a choice of 10X Reaction Buffer with Mg++ (“with Mg++”); or 10X Reaction Buffer without Mg++ and a separate tube of 25 mM MgCl2 (“separate Mg++”).
QC specifications for EconoTaq DNA Polymerase are rigorous:
Concentration: 5 units/µl. One unit catalyses the incorporation of 10 nmol of dNTP into acid-insoluble material in 30 minutes at 70°C in 50 mM Tris-HCl (pH 9.0), 50 mM NaCl, 5 mM MgCl2, 200 µM dGTP, dATP, dTTP dCTP (a mix of un-labelled and [33P] dCTP), 10 µg of activated calf thymus DNA, and 0.1 mg/ml BSA.
Storage Buffer: 10 mM Tris-HCl (pH 7.5), 100 mM KCl, 0.1% Triton X-100, 0.1 mM EDTA, 1mM DTT, and 50% glycerol.
10X Reaction Buffer:100mM Tris-HCl (pH 9.0), 500 mM KCl, 1% Triton X-100, and with or without 15mM MgCl2.
PCR Activity: EconoTaq DNA Polymerase is tested in DNA amplification using a variety of templates and primers.
Activity Determination: One unit of EconoTaq DNA Polymerase catalyses the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 minutes at 70 °C in 50 mM Tris-HCl (pH 9.0), 50 mM NaCl, 5 mM MgCl2, 200 µM dGTP, dATP, dTTP, dCTP (a mix of un-labelled and [33P]dCTP), 10 µg Activated Calf Thymus DNA, and 0.1 mg/mL BSA.
Absence of Endonuclease or Nicking Activity: Incubation of 10 U of EconoTaq DNA Polymerase with 1 µg of supercoiled pBR322 DNA for 16 hours at 70 °C results in no detectable conversion to relaxed or linear forms detectable by agarose gel electrophoresis.
Absence of Exonuclease Activity: Incubation of 10 U of EconoTaq DNA Polymerase with 1 µg of HindIII-cut lambda DNA for 16 hours at 70 °C resulted in no smearing of bands on agarose gels.
Purity: EconoTaq DNA Polymerase is >99% pure as determined by SDS PAGE. There is no detectable DNA contamination.
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