Thermostable DNA polymerase
Thermus aquaticus, recombinant, E. coli
For in vitro use only!
Unit Definition: One unit is defined as the amount of the enzyme required to catalyze the incorporation of 10 nmoles of dNTP's into an acid-insoluble form in 30 minutes at 70 °C using hering sperm DNA as substrate.
Shipping: shipped on blue ice
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Concentration: 5 units/μl
Taq Pol - red / high yield buffer is recommended for use in routine PCR reactions. The buffer system is optimized for high efficiency and gives superior amplification results in a broad range of reaction conditions with most primer-template pairs. The buffer system facilitates the incorporation of labeled or modified nucleotides into DNA. The inert red dye allows easier handling and facilitates the preparation of the master mix. Note that the ammonium based buffer contains detergents and may interfere with automated pipetting systems.
The enzyme replicates DNA at 72 °C. It catalyzes the polymerization of nucleotides into duplex DNA in 5'→3' direction in the presence of magnesium. It also possesses a 5'→3' polymerization-dependent exo-nuclease replacement activity but lacks a 3'→5' exonuclease activity.
Taq Pol - red / high yield buffer contains an inert red dye for easier handling and to facilitate the preparation of the master mix.
Taq Pol - red (red cap)
5 units/μl Taq DNA Polymerase in 20 mM Tris-HCl, 100 mM KCl,
0.1 mM EDTA, 1 mM DTT, 0.5 % Tween-20, 0.5 % Nonidet P-40, inert red dye, 50 % (v/v) Glycerol, pH 8.0 (25°C)
High Yield Buffer complete (green cap),
600 mM Tris-HCl, 150 mM (NH4)2SO4, 20 mM MgCl2,
0.05 % Tween-20, 0.05 % Nonidet P-40, pH 8.8 (25°C)
MgCl2 Stock Solution (yellow cap)
25 mM MgCl2
Recommended 50 μl PCR assay:
|5 μl||10x High Yield Buffer complete||green cap|
|200 μM||each dNTP||-|
|0.2 - 1 μM||each Primer||-|
|2 - 50 ng||template DNA||-|
|0.2 - 0.5 μl|
(1 - 2.5 units)
|Taq Pol -red||red cap|
|Fill up to 50 μl||PCR-grade water||-|
|MgCl2 Stock Solution||2 μl||4 μl||6 μl||8 μl|
|final MgCl2 conc.||3 mM||4 mM||5 mM||6 mM|
|94 °C||2 min||1x|
|denaturation||94 °C||30 sec||30x|
|annealing1)||45 - 68 °C||30 sec||30x|
|elongation2)||72 °C||30 sec - 4 min||30x|
|72 °C||2 min||1x|
For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary for each new template DNA and/or primer pair.
Availability Restriction: Exclusively distributed in Japan by Greiner BioOne