Multiplex PCR – It’s a party in my eppi!

Multiplex PCR – The V.I.P. among polymerase chain reactions

Do you know it?

The term multiplexing refers to the use of multiple fluorescent dyes to examine various elements within a sample.
Numerous fluorescent dyes with unique emission spectra are available and can be used to differentially label probes for various targets. Multiplexing allows thus different targets to be observed in parallel, adding more context to your observations and consequently providing more meaningful results.

Who invited it to the party?

The beginning was made in 1988 by a group of researchers from the Institute for Molecular Genetics, Baylor College of Medicine in Houston. Their paper „Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification“ [1] was the first to describe multiplexing PCR for molecular diagnostics. Since then, things have become more and more colorful in our labs.

Tell me more about Multiplex PCR!

Multiplex PCR is a cost-effective and rapid method of real time PCR that simultaneously saves required sample volume. However, the requirements for highly specific multiplex amplification can be a real challenge for researchers. A successful multiplex assay must meet important parameters. Among them, consistent amplification of all targets under the same solution and thermocycling conditions, while maintaining a low limit of detection for all targets. Primer dimer, cross-amplification, and off-target amplification must be minimized to avoid spurious results. Optimization of primer sequences, primer concentrations, salt concentrations, enzyme selection, and thermocycling profile to match the particular Tm is imperative.

Why should I get to know it?

Multiplex PCR offers a number of advantages that significantly improve your lab work, your process costs and your efficiency:

• Process cost savings compared to single real time PCR. Including consumables such as pipette tips, PCR tubes, enzymes and last but not least electricity, which has recently become a relevant cost factor.
  

• The processing time from primer and sample design to the result is much shorter with multiplex PCR. Several different detections can be detected in only one PCR run. This minimizes labor time costs and maximizes labor time efficiency.
  

• In general, much less samples are required, resulting in huge savings in sample resources that may be costly to prepare.

We are friends already!

Shortening the time to outcome is particularly important in many diseases such as sepsis, respiratory or gastrointestinal diseases. Therefore, we at Kilobaser are committed to providing every scientist with fast and independent access to various DNA probes. With our Kilobaser DNA & RNA Synthesizer Extended Edition, the Kilobaser MultiRack as well as the Kilobaser MultiClick Kit we enable you to save time, cost and effort.