Ever find yourself with more work than time to complete it? Multiplexing technologies may be your solution, allowing you to get more done in the lab while ensuring that you still go home at a reasonable hour. In addition to alleviating stress, new multiplexing assays, reagents, and equipment are accelerating the pace of research, allowing you to kill a manifold number of birds with just one stone.
The common analogy above refers to the ability to perform hundreds—and even thousands—of assays all at once. With new dyes and optical detectors, multiplexing meets the demands of the fervor to understand each gene of the human genome and every protein in every cell. Multiplexing methods—techniques that concurrently perform multiple probes for certain molecules—accelerate discovery and ensure reproducibility with the simultaneous assays serving as internal controls for the others. And by boosting the speed of discovery, researchers are coming closer to understanding and treating diabetes, obesity, cardiovascular disease, autoimmunity, inflammation, cancer, infection, and other diseases and conditions. Clinical labs benefit as well; picking up the pace of diagnostic testing can mean faster delivery of lifesaving treatments to patients.
Sophisticated dyes are central to following multiple probes added simultaneously to the same reaction vessel. These new dyes, tailor-made for multiplexing, provide distinct signals with less noise and more stability—even in the high temperatures of quantitative PCR. In addition, labeling several probes with different colored dyes enables you to convert simple microarray screenings or microplate assays into multiplex experiments.
Beads are most popular in the multiplexing world. Bound to several types of substrates, including oligonucleotides, proteins, antibodies, or drugs, the beads help reveal the biological function of the probes that specifically bind to certain substrates. For maximum convenience, you can purchase one of the many pre-made beads carrying cytokines, single nucleotide polymorphisms, phosphoproteins, and proteins associated with diabetes, obesity, cardiovascular disease, autoimmunity, inflammation, cancer, infection, and other diseases and conditions. Or, you can take advantage of services that produce customized beads, run the assays, analyze the results, and provide a comprehensive report.
In addition to simultaneously using probes with different dyes, using beads with unique spectral signatures greatly expands your multiplexing capacity. With the popular Luminex beads, for example, these signatures are the distinct spectral markers produced by a ratio of a red dye and an infrared fluorescent dye. Each Luminex bead carries one type of substrate. By incubating all 100 beads with cells, serum, or other types of samples, you can concurrently screen for 100 analytes, instead of performing 100 separate assays.
How’s that for cutting your workload? Beads and other products, such as those below, offer tremendous momentum for your project—and for getting you out the door at the end of the day.
Multiplexing real-time, quantitative PCR (rt, q-PCR) assays using un-optimized reporter dye sets places unusual performance and data analysis demands on today’s rt, q-PCR thermocyclers. Biosearch Technologies has developed two new series of performance-optimized dyes for multiplex real-time, q-PCR—the CAL Fluor® and Quasar® Dyes. These new dyes are high performing, low cost alternative dyes for many commonly used fluorophores, and are compatible with all probe/primer formats and major thermocyclers. See Biosearch's website for a complete list of compatible thermocyclers.
