Thermo Scientific Phusion U DNA Polymerase is a new engineered, high-fidelity enzyme developed using fusion technology. Due to a proprietary mutation in Phusion's dUTP-binding pocket, Phusion U overcomes an important limitation of proofreading enzymes - its ability to incorporate dUTP and read through the presence of uracil in the DNA template.

In addition to the ability to use uracil, Phusion U has all the superior properties of other Phusion DNA polymerases - high accuracy, high speed, the ability to amplify long amplicons up to 20 kb and is based on Affibody High specificity of hot start. These properties make Phusion U Hot Start DNA an excellent choice for critical applications such as amplification of bisulfite-converted or damaged DNA and control of residual contamination.

Phusion U Hot Start DNA Polymerase is also available in master mix and Green formats. Phusion U Hot Start PCR Master Mix (F-533S and F-533L) is a convenient 2X mix designed to minimize pipetting steps. Just the user adds templates and primers. Phusion U Green forms (F-556S and F-556L) are a combination of Phusion U Hot Start DNA Polymerase and 5X Phusion Green Buffer. The buffer contains a density reagent and two tracking dyes for direct gel loading of PCR products. This colored buffer does not interfere with Phusion U performance and is compatible with downstream applications such as DNA sequencing, ligation, and restriction enzyme digestion.

Key Features

• Accuracy—High-Fidelity DNA Polymerase (25x Taq)
• Uracil -Tolerant—Engineered to incorporate dUTP and amplify uracil-containing templates
• Specificity—Hot start to reduce non-specific amplification and primer degradation
br>• Speed—short extension time (15-30 s/kb)

Applications

• Amplification sulfite conversion of DNA
• Amplify damaged or aged DNA
• Residual contamination control
• Uracil excision (USER) based cloning method

Using Phusion DNA Polymerase
The annealing temperature design rules of Phusion DNA polymerase are different from those of many ordinary DNA polymerases (such as Taq DNA polymerase). For best results, use www.thermofisher.cn/tmcalculator.