Bioradiations Staff
Introducing Bio-Rad’s PrimePCR™ Assays and Panels for real-time PCR. Our PrimePCR™ assays are expertly designed and wet-lab validated to ensure optimal assay performance. Our unique approach aids in meeting the MIQE guidelines by thorough validation of every single assay using rigorous and state-of-the-art laboratory testing procedures. With the new PrimePCR assays, Bio-Rad offers a complete workflow solution for real-time PCR including RNA isolation kits, reverse transcription kits, real-time PCR supermix, as well as real-time PCR detection systems and data analysis software.
The PrimePCR product portfolio includes:
• Individual SYBR® Green assays
• Pre-plated pathway and disease specific panels
• Custom configured plates
• Experimental control assays
• Reference gene panels
Fig.1. The PrimePCR assays and panels. The online ordering tool enables you to browse and order primer assays and pathway panels. It also allows you to design your own custom plate or create a custom assay.
Guaranteed Performance
Wet-lab validation of every assay provides confidence in results while eliminating the time-consuming assay design and optimization steps. In accordance with the MIQE guidelines, the PrimePCR assays were evaluated in terms of specificity, efficiency (see Figure 2), linear dynamic range, and background signal in negative controls. Assays are validated for use with iScript™ Advanced cDNA synthesis kit for RT-qPCR and SsoAdvanced™ SYBR® Green supermix for superior performance. Validation information is available for every assay online at www.bio-rad.com/PrimePCR. The details of the assay performance standards are indicated in the table below.
Assay Performance Standards and Efficiencies
| Sensitivity | Accurate detection of 20 copies |
| Specificity | Validated amplicon sequence with next generation sequencing (NGS). Minimal primer dimer formation and genomic DNA cross reactivity |
| Amplification Efficiency | 90%–110% |
| Dynamic Linear Range | Minimum of six orders of magnitude. Detection of a symbolic template standard curve from 20 to 20 million copies |
| R² | >0.98 |
Fig. 2. Distribution of PrimePCR assay efficiencies. While efficiencies of 90–110% were targets, final analysis shows that a large majority of assays actually fall between 95 and 105%.
More Than 300 Pathway and Collection Panels
Predesigned real-time PCR plates are available for canonical pathways and related collections to help identify and investigate key targets in a biological area of interest. There are interactive pathway maps for 260 canonical pathways online. The pathway maps illustrate protein interactions and regulation to provide a comprehensive picture of signaling and disease processes (see Figure 3).
Fig. 3. Apoptosis and survival — TNFR1 signaling pathway map. This pathway map is one example of the 260 canonical pathway maps provided by Thomson Reuters. Pathway maps illustrate protein interactions and regulation to provide a comprehensive picture of signaling and disease processes. On the Bio-Rad website, each target on the map can be selected to view related gene and protein information.
Customizable 96- and 384-well Plate Formats
The custom plate configurator allows users to easily layout assays on a plate exactly as they choose or use a suggested plate layout template. The configurator guides the user through each step of the plate configuration process, such as selecting a plate template (see Figure 4), searching for and selecting assays, adding assays to the plate, and reviewing the plate for accuracy.
Fig. 4. PrimePCR custom plate configurator. A screenshot from the first steps of the plate configurator online.
Visit www.bio-rad.com/PrimePCR to search and order assays, panels, and controls. PrimePCR assays are available for ordering in select countries. Please check with your local Bio-Rad representative for details.