Optimizing RT-qPCR assays for multiplex gene expression analysis

26 Aug 2026

Learn how Bio-Rad solutions can help optimize RT-qPCR workflows for reliable multiplex gene expression analysis. This application note explores best practices for reference gene selection, assay validation, and data analysis, and demonstrates how Bio-Rad’s validated assays, reagents, instruments, and software can support multiplex results comparable to singleplex qPCR—helping streamline workflows without compromising data quality.

Resource details:

Resource type: Application note
Page count: 4
Read time: 6 mins

CFX Opus Real-Time PCR Systems

Bio-Rad

The CFX Opus System is the next evolution in qPCR from Bio-Rad. With improved thermal performance and our proprietary, accurate optical shuttle system, your data will be more consistent than ever. A sleek, modern design includes a refreshed and easy-to-use interface with new connectivity options and features. In addition to ethernet compatibility, the CFX Opus System can connect directly to Wi-Fi for completely wireless operation and data retrieval. The CFX Opus System is capable of stand-alone operation and direct data transfer to a USB device, or it can be connected to a computer. With internet connectivity, the CFX Opus System can be programmed to send your data and notifications directly to your email or local networked drive, enabling easy access by collaborators or from other network-connected locations. Features and Benefits Quick system set up — easy installation and factory-calibrated optics Our best thermal uniformity and accuracy to date Offers Wi-Fi connectivity and BR.io cloud platform analysis tools Sensitive modern touch screen — responsive even with double gloves Updated interface is easier to use and more intuitive than ever Uniform optical system free of regular calibration requirements Audit and e-signature compatibility with CFX Maestro Security Edition Software Available in 96-well (5-color detection) and 384-well (4-color detection) formats Applications and Uses Gene expression without requiring ROX End-point analysis for sequence detection Protein thermal shift assays Assay optimization with thermal gradient

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PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Gene Expression and Molecular CloningMolecular cloning is a set of techniques that utilizes vectors to transfer recombinant DNA into host cells and is an essential tool for investigating the expression of genes and proteins in bacterial or mammalian cells. A variety of vectors optimized for gene cloning and expression in a range of host organisms are available, alongside competent cells for genetic replication. Here, you can explore a range of molecular tools, high-quality genomic and cDNA libraries, premade clones, transformation and transfection reagents and mutagenesis or gene expression detection assays and expression arrays. Find the best gene expression and molecular cloning products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.RT qPCRReal-time quantitative PCR (RT-qPCR) is a technique used to measure gene expression and quantify DNA or RNA in real-time. It is essential in genomics, diagnostics, and biotechnology research. Explore RT-qPCR systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.