VividTaq™ DNA Polymerase

Description

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

Applications

• 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

Reagents Supplied

10X Taq Buffer A
10X 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 MgCl₂ concentration
10 x 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

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