Design Your Own Droplet Digital™ PCR Assays
Product News Aug 25, 2016
A critical aspect to the success of a ddPCR experiment is the use of a well-designed assay, which requires careful selection of PCR primers and probes specific to the gene of interest. Many factors influence assay design, such as amplicon length, GC content, sequence specificity, and melting temperature, all of which impact the final target sequence, primers, and probes. Flawed assay design can result in laborious testing and optimization, which may take several iterations.
Bio-Rad’s ddPCR assay website takes the guesswork out of ddPCR assay design and expands the portfolio of assays already available to ddPCR users. The website enables users to instantly design assays for any target of interest or order preexisting mutation detection and copy number determination assays.
The portfolio of preexisting assays includes nearly 1,000 wet-lab validated assays guaranteed to work when properly used and more than 2,500 predesigned assays that employ the same design criteria to deliver reliable and robust ddPCR results. Assays that have not been predesigned and/or validated can be designed in real time through the website and will include protocols to help users optimize their results for a high rate of success. Detailed information is provided for each assay, including the MIQE context sequence, amplicon length, compatible restriction enzymes, and recommended copy number variation reference assays.
Extending customer convenience further, Bio-Rad will also offer a combined assay format for mutation detection that includes both mutant and wild-type assays in a single tube. This format eliminates the potential for user error when assays are manually combined.
Additional benefits of Bio-Rad’s new assay design website include:
• Comprehensive analysis of assay specificity — perform sequence alignment to human, mouse, or rat reference genomes
• Multiple fluorophore options — choose either FAM or HEX probes for copy number determination assays
• Assay validation tools — order positive-control DNA for mutation detection assays