20 mM Tris-HCl (pH8.0)

0.1 mM EDTA

1 mM DTT

100 mM KCl

50% Glycerol

Stabilizers

Taq Platinum DNA Polymerase is a chemical-modified Hot Start Taq polymerase with 3'-5' exonuclease activity and 5'-3' exonuclease activity. The enzyme activity of Taq Platinum DNA Polymerase is blocked at room temperature. Its activity can only be activated after heating at 94°C for 5-10 min, thus avoiding non-specific amplification caused by primer non-specific annealing or primer dimer at low temperature before the initial cycle of PCR reaction, and greatly improving the sensitivity and specificity of PCR reaction. In addtion, Taq Platinum DNA Polymerase has very high fidelity, which is second best to Pfu polymerase. The extension speed of DNA polymerization is faster than Pfu polymerase and the amplification efficiency is higher.

One-tube Taq Platinum MasterMix (National High-Tech Product Certification)

■ The Taq Platinum MasterMix has improved specificity and sensitivity of PCR reaction and can amplify complex templates with high GC content, secondary structure and the like. As low as 2 copies of the target template can be amplified, ensuring more accurate experimental results.

■ The unique Taq Platinum MasterMix formula makes the whole reaction system very stable, and the activity will not be affected by repeated freeze-thaw or long-term storage at 4°C.

■ The stable and efficient pre-prepared PCR mixed solution can make the operation fast and simple, greatly reducing labor intensity and sampling error. High-performance PCR enhancer and optimizer are also included in the mix, which reduces the requirements on PCR conditions.

■ This product has both dye-containing and dye-free systems. Dye-containing MasterMix products can be directly electrophoresed after PCR, without adding loading buffer.

The purity by SDS-PAGE detection is more than 99%; No activity of exogenous nuclease is detected; Single-copy gene in human genome could be amplified effectively; No significant activity change when stored at room temperature for one week.

It has 5'-3' exonuclease activity and 3'-5' exonuclease activity, and its fidelity is next to Pfu polymerase. The extension speed of Taq Platinum Polymerase is faster than Pfu polymerase and the amplification efficiency is higher. PCR products can be directly ligated to the blunt end or cloned with TA vector. If the cloning efficiency needs to be improved, it is recommended to purify first and add 3’-dA overhangs before cloning into TA vector.

■ The primer length is usually 20-25 mer. However, when performing long fragment PCR, the primer length should be increased to 30-35 mer.

■ There is no complementary pairing between the two primers, especially for the last 3 bases at the 3' end.

■ GC content should be 50-60%, and avoid local rich GC or AT. In order to make primer and template bind stably, avoid AT rich structure at the 3' end.

■ Avoid primer to form secondary structure.

■ Select two primers with Tm temperatures close to each other.

■ When the primer is less than 20 mer: Tm=2°C×(A+T)+4°C×(G+C).

■ When the primer is more than 20 mer: Tm=81.5+0.41×(GC%)-600/L, where L is the length of the primer.

■ Set the annealing temperature at (Tm-5)°C.