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Taq DNA Polymerase

  • Stable thermophilic DNA polymerase, suitable for applications requiring high temperature synthesis of DNA.
  • Description:
  • Taq DNA Polymeraseis a thermostable enzyme of approximately 94 kDa from Thermus aquaticus.
  • Ultrapure, recombinant protein.
  • The enzyme replicates DNA at 74°C and exhibits a half-life of 40 min at 95°C (1,2).
  • Catalyzes the polymerization of nucleotides into duplex DNA in the 5´->3´ direction in the presence of magnesium ions.
  • Maintains the 5´->3´ exonuclease activity.
  • Lacks the 3´->5´ exonuclease activity.
  • Adds extra A at the 3′ ends.
  • Taq DNA Polymerase is recommended for use in PCR and primer extension reactions at elevated temperatures to obtain a wide range of DNA products up to 10 kb.

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmoles of dNTP into acid-insoluble material in 30 min at 74°C. The reaction conditions are: 50 mM Tris-HCl (pH 9.0 at 25°C), 50 mM NaCl, 5 mM MgCl2, 200 μM each of dATP, dCTP, dGTP, dTTP (a mix of unlabeled and [³H]dTTP), 10 μg activated calf thymus DNA and 0.1 mg/ml BSA in a final volume of 50 μl.

Storage Conditions: Store at -20°C

Storage Buffer: 20 mM Tris-HCl (pH 8.0 at 22°C), 100 mM KCl, 0.5% Tween 20, 0.5% Igepal CA-630, 0.1 mM EDTA, 1 mM dithiothreitol, 50% glycerol and stabilizers.

۱۰ x Reaction Buffer:

۱۰ x Pol Buffer A (optimization buffer without MgCl2): the buffer allows to optimize MgCl2 concentration.

۱۰ x Pol Buffer B (general application, up to 10 kb): the buffer contains 15 mM MgCl2 and is optimized for use with 0.2 mM of each dNTP.

۱۰ x Pol Buffer C (coloured): 10 x Pol Buffer B enriched with two gel tracking dyes and a gel loading reagent. The buffer enables direct loading of PCR products onto an agarose gel