i-pfu DNA Polymerase
PCR core kit of high-purity pfu DNA polymerase with low error rate and proofreading function that is essential for high-accuracy gene cloning
i-pfu DNA Polymerase is a typical heat-resistant proof-reading enzyme with 3 '→ 5' exonuclease activity. It has high fidelity and guarantees an error rate of 1x10-6 (one error rate per 13,000 base pairs) .The proof-reading activity of pfu DNA Polymerase is that when a DNA polymerase is polymerized, mismatched (non-complementary) nucleotides can be removed and corrected with complementary nucleotides. This activity allows PCR products of more accurate base sequences to be obtained than Taq DNA Polymerase, but allows amplification of longer DNA templates compared to Taq DNA Polymerase. This is because the mismatched nucleotide interferes with the polymerization reaction.
The Pfu DNA Polymerase can remove the DNA polymerase from the template during the DNA synthesis process. It is suitable for long-size PCR (up to ~ 30 kb) which is difficult to amplify. It has high fidelity and proof reading function compared with general Taq DNA Polymerase. As a result, DNA fragments longer than Taq DNA Polymerase (up to 30 kb) can be amplified. Because of these properties, Pfu DNA Polymerase is widely used in experiments where accuracy is required, such as gene cloning and expression, although polymerization efficiency is low and it takes more time to polymerize.Unlike Taq DNA Polymerase, this i-pfu DNA Polymerase produces blunt-end amplification products. Therefore, in order to clone the amplification product, it can be cloned into a blunt-end vector. To clone T vector, a base A must be added.
|i-pfu DNA Polymerase(2.5U/ml)||250 units x 1 tube|
|10× PCR Buffer(w/15 mM Mg2+)||1 ml x 1 tube|
|10× Mg2+ free PCR Buffer||1 ml x 1 tube|
|10 mM dNTPs (2.5 mM/each)||500 μl x 1 tube|
|25 mM Mg2+||1 ml x 1 tube|
Sensitivity comparison test
To compare the sensitivity of i-pfu DNA Polymeras and Company A and B using the same function product, 1 Kb and 4.5 Kb fragment primer was diluted by template concentration and amplified. i-pfu DNA Polymeras exhibits excellent sensitivity even at lower template concentrations compared to products with the same function of other companies.
[ Panel A ] 1 Kb amplification(λDNA)
Lane M, 1 Kb DNA Ladder; lane 1, 200 pg λDNA; lane 2, 100 pg λDNA; lane 3, 50 pg λDNA; lane 4, 25 pg λDNA; lane 5, 12.5 pg λDNA; lane 6, 6.25 pg λDNA; lane 7, 3.125 pg λDNA; lane N, negative control
[ Panel B ] 4.5 Kb amplification(TopoXL/5F)
Lane M, 1 Kb DNA Ladder; lane 1, 50 ng DNA; lane 2, 10 ng DNA; lane 3, 2 ng DNA; lane 4, 400 fg DNA; lane 5, 80 fg DNA; lane 6, 16 fg DNA; lane 7, 3.2 fg DNA; lane N, negative control