Taq DNA Polymerase (Top quality general application Taq DNA polymerase)

Top quality general application Taq DNA polymerase

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

Description:

• Taq DNA Polymerase is 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, 0.1 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.