Details

Description

• Perpetual OptiTaq DNA Polymerase is a modified and balanced blend containing top quality Thermus aquaticus DNA Polymerase, Pyrococcus furiosus DNA Polymerase, anti-Taq DNA Polymerase antibodies, reversible inhibitors and enhancers.
• Ultrapure, recombinant proteins are used to prepare Perpetual OptiTaq DNA Polymerase.
• Our carefully selected anti-Taq antibodies have high thermal stability, providing protection against non-specific primer extension from room temperature to 70°C.
• The polymerase activity is restored during the initial denaturation step when amplification reactions are heated at 92-95°C for two minutes.
• Formation of inactive complexes between Taq DNA Polymerase and an anti-Taq antibody forms a basis for automatic \"hot start\" PCR, which allows for the assembly of PCR reactions at room temperature.
• High stability of the complexes allows for the enormous increase of PCR specificity, sensitivity and yield in comparison to the conventional PCR assembly method.
• Automatic \"hot start\" PCR is a convenient method when assembling multiple PCR reactions, saving time and reagents.
• Clean and safe laboratory practice assured, due to removed necessity to open hot tubes.
• The blend catalyzes the polymerization of nucleotides into duplex DNA in the 5→3 direction in the presence of magnesium ions and exhibits the 3→5 proofreading activity, resulting in considerably higher PCR fidelity than possible with unmodified Taq DNA polymerase (1).
• Enables increased amplification product yield in comparison with Taq DNA polymerase over wide range of PCR products.
• Maintains the 5→3´ exonuclease activity.
• Adds extra A at the 3 ends.
• Suitable for multiplex PCR due to increased specificity, wider tolerance for Mg²⁺, salts concentration, and pH (2,3).
• Improves PCR results with critical templates, such as containing GC-rich regions, palindromes or multiple repeats.
• Perpetual OptiTaq 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 20 kb.

Storage Buffer

20 mM Tris-HCL (pH 8.0 at 22°C)100 mM KCI0.5% Igepal CA-6300.1 mM EDTA1 mM dithiothreitol50% glycerol

10X Reaction Buffers

10X Pol Buffer A (optimization buffer without MgCl₂)The buffer allows to optimize MgCl₂ concentration.

10X Pol Buffer B (general application without MgCl₂)The buffer contains 15 mM MgCl₂ and is optimized for use with 0.2 mM of each dNTP.

10X Pol Buffer C (colored)10X 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 singe- and double-stranded DNase activities. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis.

References

(1) Cline, J., Braham, J. and Hogrefe, H. (1996) Nucleic Acids Res. 24, 3545.(2) Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bacteriol. 127, 1550.(3) Kaledin, A.S., Silusarenko, A.G. and Gorodetskii, S.I. (1980) Biokhimiya 45, 644.