Description
Taq Plus DNA Polymerase Recombinant is a mixture of Taq and Pfu. Taq Plus is used to improve the reliability and yield of conventional primer extension reaction.
Taq Plus has two following advantages over Taq:
- High fidelity with an error frequency 1.6/106 (or 0.0016/103) during DNA synthesis.
- Taq Plus increases the efficiency of polymerization reaction, resulting in a great percentage of extenuation reaction completion up to 10 kb to 30 kb. Pfu has a temperature optimum between 72-78°C and remains > 95% active following 1-hour incubation at 95°C.
Source
Recombinantly produced in E. coli.
Unit Definition
One unit of the enzyme catalyzes the incorporation of 10nmole of deoxyribonucleotides into a polynucleotide fraction in 30 min at 74°C.
Storage Buffer
20 mM Tris-HCl (pH 8.0), 1 mM DTT, 0.1 mM EDTA, 100 mM KCl, Stabilizers, and 50% glycerol.
Formulation
5U/µl.
10x Reaction Buffer
200 mM TrisHCI (pH 8.8)
100 mM KCI
100 mM (NH4)2SO4
20 mM Mg SO4
1% Triton X-100
1 mg/ml bovine serum albumin (BSA).
Reaction Conditions
All reagents, including Taq Plus, should be mixed immediately before use. DNA synthesis is performed in 100 ul of mixture containing 20‐200 uM dNTPs, 0.3‐1 uM Primers, 0.1‐ 0.250 ng of template DNA, 10 ul of 10x reaction buffer and 2.5‐5 units of Taq Plus. Mix the reaction gently, centrifuge briefly and then overlay with light mineral oil. Inially, denature the reacon by incubang at 95°C for 5 minutes and then cool to 40‐68°C for 5 minutes to allow the primers to anneal to the template DNA.
Optimization of DNA Synthesis
It is important to ad the reaction components in the following order:
- H2O.
- 10x Reaction Buffer.
- dNTPs.
- DNA template and primers.
- Taq Plus.
Shipping & Storage
Ships on ice packs. Upon receipt, store the product at the temperature recommended below:
Stable for 5 days at 10°C, for longer period of time store at -20°C.
Usage
This product is intended for Laboratory Research Use Only. Not for use in diagnostic or therapeutic procedures. This product may not be used as a pharmaceutical or veterinary drug, agricultural product, or food additive.