Multiplex PCR, or multiplex polymerase chain reaction, is a variation of the traditional PCR method that allows for the amplification of multiple DNA targets in a single tube. Multiplex PCR can concurrently amplify multiple target DNA sequences in a single reaction, while standard PCR can only amplify one target sequence. Therefore, multiplex PCR offers the advantage of time and resource savings.

Thermo Scientific Phusion U Multiplex PCR Master Mix facilitates multiplex PCR by amplifying 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. The master mix contains all the components for end-point multiplex PCR except for primers and DNA template and requires minimal optimization of reaction conditions.


Thermo Scientific Phusion U Multiplex PCR formats


Benefits of Phusion U Multiplex PCR Master Mix

  • 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


Benchmarking data on Phusion U Multiplex PCR Master Mix

The Phusion U Multiplex Master Mix enables robust amplification of all amplicons in highly multiplexed reactions. Over 20 targets up to 2.5 kb can be amplified in a single tube (Figure 1).

Gel with flanking marker ladders and and multiple lanes of samples

Figure 1. Multiplex capabilities: 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%. M1: Thermo Scientific GeneRuler 50 bp DNA Ladder, M2: Thermo Scientific GeneRuler 100 bp Plus DNA Ladder

Phusion U Multiplex PCR Master Mix enabled amplification of multiple fragments with a fast PCR protocol offering excellent performance and speed.

Gel with flanking marker ladders and and multiple lanes of samples

Figure 2. 19 fragments (73–2,527 bp) from human genomic DNA were simultaneously amplified using different multiplex PCR master mixes according to manufacturers’ recommendations. M: GeneRuler 100 bp Plus DNA Ladder

Phusion U Multiplex PCR Master Mix enabled concurrent amplification of multiple targets with low quantities of template DNA.

Composite of four gels; each gel has five sample lanes and one ladder lane

Figure 3. 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

Phusion U Multiplex PCR Master Mix exhibits high tolerance to PCR inhibitors, such as blood, and enables robust amplification of all targets.

Gel with six lanes of sample flanked on either side by ladders

Figure 4. 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. The Phusion U Multiplex PCR Master Mix delivered best results. M: GeneRuler 100 bp Plus DNA Ladder

Highly multiplexed PCR

The Phusion U Multiplex Master Mix enables robust amplification of all amplicons in highly multiplexed reactions. Over 20 targets up to 2.5 kb can be amplified in a single tube (Figure 1).

Gel with flanking marker ladders and and multiple lanes of samples

Figure 1. Multiplex capabilities: 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%. M1: Thermo Scientific GeneRuler 50 bp DNA Ladder, M2: Thermo Scientific GeneRuler 100 bp Plus DNA Ladder

Exceptional performance

Phusion U Multiplex PCR Master Mix enabled amplification of multiple fragments with a fast PCR protocol offering excellent performance and speed.

Gel with flanking marker ladders and and multiple lanes of samples

Figure 2. 19 fragments (73–2,527 bp) from human genomic DNA were simultaneously amplified using different multiplex PCR master mixes according to manufacturers’ recommendations. M: GeneRuler 100 bp Plus DNA Ladder

Highly sensitive PCR

Phusion U Multiplex PCR Master Mix enabled concurrent amplification of multiple targets with low quantities of template DNA.

Composite of four gels; each gel has five sample lanes and one ladder lane

Figure 3. 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

Phusion U Multiplex PCR Master Mix exhibits high tolerance to PCR inhibitors, such as blood, and enables robust amplification of all targets.

Gel with six lanes of sample flanked on either side by ladders

Figure 4. 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. The Phusion U Multiplex PCR Master Mix delivered best results. M: GeneRuler 100 bp Plus DNA Ladder


Related brochures

 PCR and electrophoresis workflow
 Molecular biology research essentials

Additional resources

Stylesheet for Classic Wide Template adjustments

仅供科研使用，不可用于诊断目的。

仅供科研使用，不可用于诊断目的。