DNA Polymerase/Master Mixes

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 proofreading activity of i-pfu DNA Polymerase allows PCR products with more accurate base sequences to be obtained than when using Taq DNA Polymerase. When using i-pfu DNA Polymerase, DNA fragments longer than Taq DNA Polymerase (up to 30 kb) can be amplified. Because of these properties, i-pfu DNA Polymerase is widely used in experiments in which high accuracy is required, such as gene cloning and expression. Unlike Taq DNA Polymerase, this i-pfu DNA Polymerase produces blunt-end amplification products.

Sensitivity Comparison

Figure 1: To compare the sensitivity of i-pfu DNA Polymeras with similar products from Company A and B, 1 Kb and 4.5 Kb fragment primers were diluted by template concentration and amplified. i-pfu DNA Polymeras showed excellent sensitivity even at lower template concentrations compared to other products with the same function.

[ 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

Features

• High accuracy makes it possible to amplify long DNA templates 
• PCR products have more accurate bases 
• Low error rate of 1x10^-6
• Used in blunt-end Cloning
• Ideal for experiments requiring high accuracy such as gene cloning and expression

Applications

• General PCR
• Blunt Cloning
• Template amplification for DNA sequence analysis

Contents

• i-pfu DNA Polymerase (2.5 U/ml), 250 units
• 10× PCR Buffer (with 15 mM Mg²⁺), 1ml
• 10× Mg²⁺-free PCR Buffer, 1ml
• 10mM dNTPs (2.5 mM/each), 500μl 
• 25mM Mg²⁺, 1ml

Literature