Multiplex PCR enables researchers to obtain more answers in fewer reactions and a shorter timespan than singleplex PCR, because many targets are amplified in a single PCR reaction. This makes multiplexing especially valuable in applications where reaction cost or sample availability is limiting, including mutation detection, pathogen identification, clinical diagnostics research, and infectious disease screening.
Speed and efficiency are important factors to consider when developing molecular diagnostic assays, in order to process large numbers of samples and provide actionable results in a timely manner. Every improvement in the workflow leads to a significant time savings, and eliminating pipetting steps has the added benefits of reducing the potential for contamination and error and minimizing repetitive strain-induced injury. Simplifying procedures also enables better standardization between users, which can be crucial when screening projects are scaled up for widespread use.
However, while multiplex PCR reactions are very powerful, they are not easy to set up. They require lengthy optimization—and the higher the multiplex, the more complex the setup. Other key challenges include primer design and finding PCR conditions that work for all primer pairs (which usually have different amplification efficiencies). To simplify multiplex PCR assays and streamline their workflows, we have developed a new PCR master mix—SuperPlex Premix—which combines the power of multiplex PCR with the convenience of a master mix. SuperPlex Premix is a hot-start 2X PCR master mix based on our high-yield Titanium Taq DNA polymerase, which is ideal for multiplex PCR as well as for high-yield singleplex PCR. The premix contains all the reagents necessary for multiplex PCR and is easy to use—just add primers, sample, and water.
Here, we have compared the performance of our SuperPlex Premix to four other commercially available PCR master mixes that are marketed for multiplex PCR. Our results show that SuperPlex Premix provides the most even amplification in the shortest amount of time compared to the other multiplex master mixes tested.