First used as a method in detecting deletions in the dystrophin gene back in 1988, multiplex PCR has since made its way into many protocols across a variety of research areas. While still used in gene deletion analyses, multiplex PCR, as well as qPCR, is an incredibly useful tool in many applications, such as SNP genotyping, pathogen detection, and food analysis to name just a few.
Multiplex allows for the researcher to detect multiple targets in a single reaction well. The prerequisite for this is using more than one primer pair for simultaneous target amplification and analysis.
Here are some of the key benefits you will see when adopting multiplexing into your workflow:
Saves money
Saves rare samples
More reliable results
Multiplex PCR is a useful tool in any laboratory working with repeating samples and targets. Incorporating it into your workflow allows you to work through your samples faster while using fewer samples as well as consumables. Multiplexing can be applied both in endpoint PCR as well as in real-time qPCR.
In endpoint PCR, gene targets are discriminated by amplicon size and detected typically via gel electrophoresis. The amplicon size may be limited by the properties of a DNA polymerase. Typical Taq DNA polymerases allow amplification of up to 5 kb fragments. With endpoint PCR generally, more targets can be detected in one reaction compared to real-time qPCR. In qPCR, the amplicon length is rather limited, typically between 100-200 bp, and the amplicons are detected in real-time using mostly target-specific hydrolysis probes (i.e TaqMan) or dsDNA intercalating dyes (i.e. SYBR® Green, EvaGreen®, or other similar dyes). Multiplex qPCR with hydrolysis probes is highly adopted in the diagnostic sector. It is used for detecting several pathogens and internal control in the same sample in an extremely specific manner. Its time-saving, cost-efficient, and increased reliability features make it highly beneficial for routine and high throughput experiments.
Each target gene must bind a differently labeled probe to set its amplification signal apart from the other targets in the same reaction, as each label emits fluorescent light at a different wavelength that is detected by specific channels in the qPCR cycler.
When starting your next project, consider leveraging the benefits of multiplexing to increase your efficiency!
We are proud to present Solis Hot Start HiFi DNA Polymerase, a high-performance, high-fidelity enzyme designed for precision-demanding PCR applications such as sequencing, cloning, and mutagenesis. This product has superior performance with a variety of amplicons, can be used for long-range and challenging PCR, and has broad multiplexing capabilities.
It doesn’t happen often that products developed almost three decades ago remain not only on the product list but as one of the most popular products overall. This is, however, the case with our main polymerases. These have become classics that seem to never go out of style like jeans or a little black dress. So, what makes them so special? Let’s take a look!
Every company has to start from somewhere. There can either be an “aha” moment - a sudden spark of inspiration that sets everything in motion. Other times it can happen gradually with ideas coming and going until something bigger grows out of all this brainstorming. For Solis BioDyne, it was the latter. As we celebrate 30 years since our founding, we think it’s the perfect time to share our origin story with you.
Have you ever thought about doing a PCR experiment, but you are not quite sure what product to use? Or the product you have chosen doesn’t work as well as it should and you are looking for an alternative? We are here to help you out with some recommendations from fellow scientists based on different application possibilities.
