VividTaq™ DNA Polymerase

Reagents Supplied

10X Taq Buffer A
10X Taq Buffer B

Description

• VividTaq™ DNA Polymerase is a thermostable enzyme of approximately 94 kDa from Thermus aquaticus
• Ultrapure, Recombinant protein

 Applications

• VividTaq™ 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
• 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.

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): 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

Storage Buffer

20 mM Tris-HCl (pH 8.0 at 22°C
100 mM KCl
0.1 mM EDTA
1 mM dithiothreitol
50% (v/v) glycerol
Stablizers 

Assay Conditions

50 mM Tris-HCl (pH 9.0 at 25°C)
50 mM NaCl
50 mM MgCl₂ 
200 µM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [³H]dTTP
10 µg activated calf thymus DNA
0.1 mg/ml bovine serum albumin
Final 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°C
Product 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