photo of Phusion U Multiplex PCR Master Mix

Thermo Scientific Phusion U Multiplex PCR Master Mix enables simultaneous amplification of over 20 DNA targets up to 2.5 kb over a wide range of template concentration and GC content. The Master Mix is based on the high-fidelity Phusion U Hot Start DNA Polymerase, an enzyme developed using fusion protein technology that enables fast, specific and robust multiplex PCR. This high performance Phusion PCR Master Mix contains all of the components for end-point multiplex PCR except for primers and DNA template, and requires minimal optimization of reaction conditions.

Features

  • Simultaneous amplification of over 20 targets up to 2.5 kb in length
  • Excellent results even with GC-rich or suboptimal purity templates
  • Minimal optimization of reaction conditions required
  • Direct loading of PCR products on gels with Green Buffer formats

Did you know?

The Green Buffer in the Phusion U Green Multiplex PCR Master Mix includes a density reagent and two tracking dyes, enables direct loading of PCR products on gels, and thus simplifies workflows. The Green Buffer does not affect PCR performance.

gel image of 5-, 10-, 15-, 20- and 24-plex reaction from 20 ng of human genomic DNA using Phusion U Multiplex PCR Master Mix

Highly multiplexed PCR: Over 20 targets up to 2.5 kb in a single tube

5-, 10-, 15-, 20-, and 24-plex reaction from 20 ng of human genomic DNA using Phusion U Multiplex PCR Master Mix. Size range of amplicons was from 73 bp to 2.5 kb, and GC content varied in a range of 35–67%. The Master Mix enabled robust amplification of all amplicons even in highly multiplexed reactions.

M1 – Thermo Scientific GeneRuler 50 bp DNA Ladder
M2 – Thermo Scientific GeneRuler 100 bp Plus DNA Ladder

gel image showing 19 fragments (73-2527 bp) from human genomic DNA simultaneously amplified using different multiplex PCR master mixes

Unmatched performance and speed

19 fragments (73–2,527 bp) from human genomic DNA were simultaneously amplified using different multiplex PCR master mixes according to manufacturers’ recommendations. Phusion U Multiplex PCR Master Mix enabled amplification of all 19 fragments and resulted in the fastest PCR protocol.

M – GeneRuler 100 bp Plus DNA Ladder

gel image showing amplification of 14 targets (70–1000 bp) from decreasing amounts of human genomic DNA

Multiplex PCR with low amount of template DNA

Sensitivity of several multiplex PCR master mixes was tested by amplifying 14 targets (70–1,000 bp) from decreasing amounts of  human genomic DNA. Only Phusion U Multiplex PCR Master Mix delivered efficient amplification of all targets even from the smallest (0.1 ng) DNA input.

M – Thermo Scientific GeneRuler 100 bp DNA Ladder.

gel image showing 14 fragments (70–1000 bp) amplified directly from 2 µL of human blood using different multiplex PCR master mixes

Multiplex PCR with whole blood samples

14 fragments (70–1,000 bp) were amplified directly from 2 µL of human blood using different multiplex PCR master mixes according to manufacturers’ recommendations. Phusion U Multiplex PCR Master Mix enabled robust amplification of all targets from blood due to its high tolerance to PCR inhibitors.

M – GeneRuler 100 bp Plus DNA Ladder

Over 20 targets in 1 tube
gel image of 5-, 10-, 15-, 20- and 24-plex reaction from 20 ng of human genomic DNA using Phusion U Multiplex PCR Master Mix

Highly multiplexed PCR: Over 20 targets up to 2.5 kb in a single tube

5-, 10-, 15-, 20-, and 24-plex reaction from 20 ng of human genomic DNA using Phusion U Multiplex PCR Master Mix. Size range of amplicons was from 73 bp to 2.5 kb, and GC content varied in a range of 35–67%. The Master Mix enabled robust amplification of all amplicons even in highly multiplexed reactions.

M1 – Thermo Scientific GeneRuler 50 bp DNA Ladder
M2 – Thermo Scientific GeneRuler 100 bp Plus DNA Ladder

Unmatched performance
gel image showing 19 fragments (73-2527 bp) from human genomic DNA simultaneously amplified using different multiplex PCR master mixes

Unmatched performance and speed

19 fragments (73–2,527 bp) from human genomic DNA were simultaneously amplified using different multiplex PCR master mixes according to manufacturers’ recommendations. Phusion U Multiplex PCR Master Mix enabled amplification of all 19 fragments and resulted in the fastest PCR protocol.

M – GeneRuler 100 bp Plus DNA Ladder

Highly sensitive PCR
gel image showing amplification of 14 targets (70–1000 bp) from decreasing amounts of human genomic DNA

Multiplex PCR with low amount of template DNA

Sensitivity of several multiplex PCR master mixes was tested by amplifying 14 targets (70–1,000 bp) from decreasing amounts of  human genomic DNA. Only Phusion U Multiplex PCR Master Mix delivered efficient amplification of all targets even from the smallest (0.1 ng) DNA input.

M – Thermo Scientific GeneRuler 100 bp DNA Ladder.

High inhibitor tolerance
gel image showing 14 fragments (70–1000 bp) amplified directly from 2 µL of human blood using different multiplex PCR master mixes

Multiplex PCR with whole blood samples

14 fragments (70–1,000 bp) were amplified directly from 2 µL of human blood using different multiplex PCR master mixes according to manufacturers’ recommendations. Phusion U Multiplex PCR Master Mix enabled robust amplification of all targets from blood due to its high tolerance to PCR inhibitors.

M – GeneRuler 100 bp Plus DNA Ladder

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