Details

Description

• Thermostable enzyme of approximately 94 kDa fromThermus aquaticus
• Ultrapure, Recombinant protein

Applications

• Recommended for use in PCR and primer extension reactions at elevated temperatures to obtain a wide range of DNA productsup to 10 Kb
• Replicates DNA at 74°C and exhibits a half-life of 40 minutes 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

Advantages

• Helps visualize the addition of the polymerase to the reaction
• Confirms complete mixing
• Enables direct loading of PCR products onto an agarose gel without the addition of a gel loading buffer
• Added dyes allow tracking of electrophoresis progress
• Does not interfere with PCR performance
• Does not interfere with most downstream applications

Note: Not recommended for any applications using absorbance or fluorescence excitation

Reagents Supplied

10X Taq Buffer A10X Taq Buffer B

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 at 74°C in 30 minutes

Reaction Buffer

10 x Taq Buffer A (optimization buffer without MgCl₂):allows to optimize MgCl₂concentration10 x Taq Buffer B (general application, up to 10 kb):contains 15 mMMgCl₂and isoptimized for use with 0.2mM of each dNTP

Storage Buffer

20 mM Tris-HCl (pH 8.0 at 22°C100 mM KCl0.1 mM EDTA1 mM dithiothreitol50% (v/v) glycerolStablizers

Assay Conditions

50 mM Tris-HCl (pH 9.0 at 25°C)50 mM NaCl50 mM MgCl₂200 µM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [³H]dTTP10 µg activated calf thymus DNA0.1 mg/ml bovine serum albuminFinal volume of 50 µl

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

Storage Conditions

Store at -20°CProduct shipped on dry ice

References

1. Chien, A., Edgar, D.B. and Trela, J.M. (1976) J. Bacteriol. 127, 15502. Kaledin, A.S., Sliusarenko, A.G. and Gorodetskii, S.I. (1980) Biokhimiya 45, 644

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