Taq DNA Polymerase, Native

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Reagents Supplied

10X Taq Buffer A
10X Taq Buffer B
10X Taq Buffer C 

Source 

Thermus aquaticus

Description

• Taq DNA Polymerase is a thermostable enzyme of approximately 94 kDa from Thermus aquaticus

Applications

• The enzyme 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
• 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 of E.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 minutes at 70°C

10 x Reaction Buffer

10 x Taq Buffer A (optimization buffer without MgCl₂): the buffer allows to optimize MgCl₂ concentration
10 x Taq 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 Taq Buffer C (colored): 10 x Taq 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

Assay Conditions

25 mM Tris-HCl (pH 9.5 at 25°C)
50 mM KCl
10 mM MgCl₂
1 mM dithiothreitol 
200 μM each of dCTP, dGTP, dTTP, and dATP (a mix of unlabeled and [α-³²P] dATP)
10 μg activated calf thymus DNA
1 mg/ml bovine serum albumin 
15 µg activated calf thymus DNA
Total reaction volume is 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°C
Shipped on dry ice 

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