Thermo Scientific Phusion U Multiplex PCR Master Mix is a ready-to-use, 2X end-point PCR master mix designed for simultaneous amplification of multiple targets in a single tube. Over 20 primer pairs may be combined into a single reaction for highly specific and efficient multiplexing over a broad range of primer and template concentrations.

The Phusion U Multiplex PCR Master Mix contains a proprietary buffer with balanced concentrations of all PCR components eliminating the need for tedious optimization. The Phusion U Green Multiplex PCR Master Mix (F-564S or F-564L) further simplifies the workflow - it includes a density reagent and two electrophoresis tracking dyes for direct loading of PCR products on gels.

Highlights
• Specific and efficient multiplex PCR of over 20 targets up to 2.5 kb
• Excellent results with templates of suboptimal purity due to inhibitor tolerance
• Fast cycling and direct loading of PCR products on gels with Green format (F-564S and F-564L)

Applications
• Genotyping
• Pathogen detection
• Food testing
• Analysis of genetically modified organisms
• Amplification of microsatellites

The master mix is based on Phusion U Hot Start DNA Polymerase, a high performance enzyme developed using fusion technology. Similarly to other enzymes from Phusion family, Phusion U DNA Polymerase incorporates more nucleotides per binding event than any other DNA polymerase. This allows achieving high yields of PCR products with shorter extention times and consequently reduced total protocol times. Significantly enhanced processivity also enables successful multiplex PCR on difficult targets such as GC-rich templates or templates of suboptimal purity. Being highly tolerant of many PCR inhibitors, Phusion U Multiplex PCR Master Mix can even be used with unpurified samples such as blood or serum.

Specificity of multiplex PCR is increased by Affibody-mediated hot start mechanism. Reversibly bound Affibody ligand inhibits the polymerase at ambient temperatures preventing amplification of nonspecific products and formation of primer-dimers.

Using Phusion DNA Polymerases
Annealing rules for Phusion DNA Polymerases are different from many common DNA polymerases (such as Taq DNA polymerases). For optimal results, use our Tm calculator at www.thermofisher.com/tmcalculator.