Native Taq DNA Polymerase

Stable thermophilic DNA polymerase, suitable for applications requiring high temperature synthesis of DNA.

Source: Thermus aquaticus

Description:

• Taq DNA Polymerase is a thermostable enzyme of approximately 94 kDa from Thermus aquaticus.
• 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.
• 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.
• Native Taq DNA Polymerase is recommended for use in special PCR applications, where traces ofE.coli genomic DNA may interfere with amplification specificity.

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, 0.1 mM EDTA, 1 mM dithiothreitol, 50% glycerol and stabilizers.

10 x Reaction Buffer:

10 x Pol Buffer A (optimization buffer without MgCl₂): the buffer allows to optimize MgCl₂concentration.

10 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.

10 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.

Quality Control: All preparations are assayed for contaminating endonuclease, 3'-exonuclease, and nonspecific single- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.

References:

1. Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bacteriol. 127, 1550.
2. Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I.(1980) Biokhimiya 45, 644.