50 mM Tris-HCl (pH8.2)

0.1 mM EDTA

1 mM DTT

50% Glycerol

Stabilizers

Pfu DNA Polymerase is expressed from E.coli with cloned Pyrococus Furiosis DNA Polymerase gene and purified and separated by multiple column purification. Because Pfu has 3'-5' exonuclease activity, it can proofread in DNA amplification process, while traditional Taq DNA Polymerase cannot. Although other Taq DNA polymerases such as Vent, Deep Vent, Tli, UITma, etc. have proofreading functions, Pfu has the lowest mismatch rate among all Taq DNA polymerases found so far. Pfu DNA Polymerase has better thermal stability than common Taq DNA polymerase, and it can maintain more than 90% activity at 95°C for 1 hour. 

One-tube Pfu PCR Mix (China National High-Tech Product Certification)

■ The Pfu PCR Mix 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 Pfu 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 mix 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 PCR Mix products can be directly electrophoresed after PCR, without adding sample buffer.

The activity of 1 unit (U) Pfu DNA Polymerase is defined as the amount of enzyme required to incorporate 10 nmol deoxynucleotides into acid-insoluble substances at 74°C within 30 min using activated salmon sperm DNA as a template/primer.

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 3'-5' exonuclease activity and no 5'-3' exonuclease activity. The extension speed of DNA amplification is lower than that of Taq polymerase, and generally the extension speed of Pfu enzyme is 0.5-1 kb per minute. The thermal stability of Pfu is better than Taq. For templates with high GC content, the denaturation temperature can be increased to 98°C, which has no effect on the activity of Pfu polymerase. The PCR product is blunt-ended, which can be added with 3’-dA overhangs before ligated with TA vector or cloned with blunt-ended vector.

It can be used for high fidelity amplification of DNA, such as gene expression cloning, site-directed mutation, single nucleotide polymorphism (SNP) analysis and end repairing.