Taq DNA Polymerase, Native

Source 

Thermus aquaticus

Description

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

Applications

• 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
• Recommended for use in PCR and primer extension reactions at elevated temperatures to obtain a wide range of DNA products up to 10 Kb
• Recommended for use in special PCR applications, where traces of E.coli genomic DNA may interfere with amplification specificity

Reagents Supplied

10X Taq Buffer A
10X Taq Buffer B
10X 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): Allows to optimize MgCl₂ concentration
10X Taq Buffer B (general application, up to 10 kb): Contains 15 mM MgCl₂ and is optimized for use with 0.2 mM of each dNTP
10X Taq Buffer C (colored): Enriched with two gel tracking dyes and a gel loading reagent, 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