Related Product Information
Taq DNA Polymerase
retains full activity after incubation at 95°C for 4 hours and has five-fold better processivity than Taq DNA polymerase. An optimized buffer reduces the need for additional optimization in many cases, making ThermalAce™ the enzyme of choice for a wide variety of applications. Sufficient reagents are provided for 100 or 500 amplification reactions of 50 μl each (at 2 units of ThermalAce™ DNA Polymerase per reaction).
|ThermalAce™ DNA Polymerase (2 U/μl)
|10X ThermalAce™ Buffer
|50X dNTP Mix (10 mM each of dATP,
dCTP, dGTP, dTTP, at pH 8.0) 200 μl 1 ml
ThermalAce™ DNA Polymerase
2 U/μl in 50 mM Tris-HCl (pH 8.0), 100 mM KCl, 1 mM Dithiothreitol (DTT), 0.1 mM EDTA, 50% Glycerol, 0.1% Triton® X-100
10X ThermalAce™ Buffer
600 mM Tris-HCl (pH 9.25), 15 mM MgSO4, 300 mM NaCl, 0.1 mg/ml bovine serum albumin (BSA), 0.1% Triton® X-100, and proprietary components
One unit of enzyme is the amount of enzyme required to incorporate 10 nmoles of dNTPs into acid insoluble material in 30 minutes at 74°C. This is the standard unit definition for all thermostable polymerases used for PCR.
Add components in the following order to each reaction vessel on ice.
DNA template x μl Primers (100 ng each) 1 μl 50X dNTP Mix (10 mM each dNTP) 1 μl 10X ThermalAce™ Buffer 5 μl Sterile Water to 49 μl ThermalAce™ (2 U/μl)*
Note: Up to 3 U of enzyme (1.5 μl) may be added for difficult templates. A master mix can be prepared for multiple reactions to enable accurate pipetting.
Cap/seal the reaction vessels and flick with your finger for several seconds to mix. Place reaction(s) on ice until ready to cycle.
Program the thermal cycler as follows. Note that the annealing temperature may vary depending on the Tm of your primers. The optimal annealing temperature is typically 5°C below the Tm of the primers.
Denaturation 98°C 95°C
Denaturation 98°C 95°C
Annealing 65°C (5°C < Tm) 55°C (5°C < Tm)
Extension 72°C 74°C
- Maintain the reaction at 4°C after cycling. The samples can be stored at -20°C until use. Analyze 5–10 μl of sample by agarose gel electrophoresis.
- Barnes, W. M. (1992). “The Fidelity of Taq Polymerase Catalyzing PCR is Improved by an N-terminal Deletion.” Gene 112: 29–35.
- Barnes, W. M. (1994). “PCR Amplification of Up to 35-kb DNA with High Fidelity and High Yield from Lambda Bacteriophage Templates.” Proc. Natl. Acad. Sci. USA 91: 2216–2220.