Details

Source

Thermus aquaticus

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
• Taq DNA Polymerase enzyme replicates DNA at 74°C and exhibits a half-life of 40 minutes at 95°C (1, 2)
• Taq DNA Polymerase 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

Reagents Supplied

10X Taq Buffer A10X Taq Buffer B10X Taq Buffer C

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 minutes at 70°C

10X Reaction Buffer

10X Taq Buffer A (optimization buffer without MgCl₂): Buffer allows to optimize MgCl₂concentration10X Taq Buffer B (general application, up to 10 kb): Buffer contains 15 mMMgCl₂and isoptimized for use with 0.2mM of each dNTP10X Taq Buffer C (colored):10X Taq Buffer B Enriched with two gel tracking dyes and a gel loading reagent; Enables direct loading of PCR products onto an agarose gel

Storage Buffer

20 mM Tris-HCl (pH 8.0 at 22°C)100 mM KCl0.1 mM EDTA1 mM dithiothreitol50% glycerolStabilizers

Assay Conditions

25 mM Tris-HCl (pH 9.5 at 25°C)50 mM KCl10 mM MgCl₂1 mM dithiothreitol200 μM each of dCTP, dGTP, dTTP, and dATP (a mix of unlabeled and [α-³²P] dATP)10 μg activated calf thymus DNA1 mg/ml bovine serum albumin15µg activated calf thymus DNATotal reaction volume is 50µl

Quality Control

All preparations are assayed for contaminating endonuclease, 3-exonuclease, and nonspecific single- and double-stranded DNase activitiesTypical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis

Storage Conditions

Store at -20°CShipped on dry ice

Downloads

MSDS PDF

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