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We have collaborated with Biogazelle, leaders in real-time PCR, to design and experimentally validate SYBR® Green-based gene expression assays for the human and mouse transcriptome. Primer pairs for each gene were designed by prioritizing gene regions most commonly occurring in transcript variants. Every assay was wet-lab validated using iScript™ advanced cDNA synthesis kit and SsoAdvanced™ SYBR® Green supermix.
Assay Performance Standards
PrimePCR Assays: Meeting the MIQE Guidelines by Full Wet-lab Validation
Bio-Rad Laboratories has partnered with GeneGo, a Thomson Reuters business and leading provider of systems biology tools, to offer a complete pathway solution for real-time PCR-based gene expression analysis. As part of the collaboration, Thomson Reuters provided interactive pathway maps for 260 canonical pathways. Each pathway belongs to one or more general biological categories such as Cancer. The pathway maps illustrate protein interactions and regulation to provide a comprehensive picture of signaling and disease processes. The curated pathways were also used to design panels of real-time PCR assays specifically tailored to the top ranked gene targets for differential gene expression analysis. Each gene target within a pathway was assigned a score based on frequency of differential expression and popularity in scientific research both historically and currently. The resulting score were used to rank and select which assays were included on the corresponding real-time PCR pathway panel.
PrimePCR Pathway Analysis: Pathway Curation and Real-Time PCR Panel Design Strategy
Control assays and synthetic DNA templates are designed to aid in assessing the key experimental factors that may be impacting your real-time PCR results.
DNA Contamination Control AssayThe PrimePCR™ DNA contamination control assay is designed to determine if genomic DNA (gDNA) is present in a sample at a level that may affect PCR results. The assay may also be used to compare relative levels of gDNA contamination present in different samples to determine if PCR results may be affected.
Positive PCR Control AssayThe PrimePCR positive control assay is designed to qualitatively assess the performance of a PCR reaction associated with a single sample. The assay may also be used to compare the relative performance of the PCR reactions associated with different samples.
RNA Quality AssayThe PrimePCR RNA quality assay is designed to determine if RNA integrity may adversely affect PCR results of a single sample. The assay may also be used to compare the relative RNA integrity between different samples in order to determine if PCR results may be affected.
Reverse Transcription Control AssayThe PrimePCR reverse transcription control assay is designed to qualitatively assess the performance of the reverse transcription reaction associated with a single sample. The assay may also be used to compare the relative performance of the reverse transcription reactions associated with different samples.
Reference Gene AssaysReference genes are used in relative gene expression analysis to normalize for variation in the amount of input messenger RNA (mRNA) between samples. For accurate quantitation, it is important to use one or more reference genes that are shown not to change expression as a result of the experimental treatment or condition. To aid in this, we have suggested a set of commonly used reference genes that can be used individually, easily screened using our pre-plated 96-well and 384-well reference panels, or added to custom designed plates.
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Aurum™ Total RNA Mini KitThe Aurum total RNA mini kit produces high-quality DNA-free total RNA from a wide range of starting materials including cultured cells, bacteria, and yeast, as well as animal and plant tissues.
iScript™ Advanced cDNA Synthesis Kit for RT-qPCRThe iScript advanced cDNA synthesis kit for RT-qPCR is a simple, fast, and sensitive first-strand cDNA synthesis kit for gene expression analysis by reverse-transcription qPCR.
PrimePCR™ Assays and PanelsExperimentally validated real-time PCR assays are available as individual assays, preplated pathway- and disease-specific panels, or custom- configured plates.
SsoAdvanced™ Universal Real-Time PCR SupermixThe SsoAdvanced™ universal real-time PCR supermixes employ our patented Sso7d fusion enzyme technology for higher processivity, increased PCR inhibitor tolerance, and robust performance with difficult qPCR targets. Available for probe- and dye-based assays in ROX- dependent or ROX-independent real-time PCR detection systems.
Real-Time PCR Detection SystemsReal-time PCR amplification systems combine thermal cyclers with optical reaction modules for singleplex and multiplex detection of fluorophores. All systems feature thermal gradient functionality.
Amplification SoftwareSoftware for Bio-Rad thermal cyclers supports multi-instrument control and collection and analysis of real-time data. Data analysis tools include ΔCq and ΔΔCq analyses and scatter plot, volcano plot, heat map, and clustergram outputs.
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View the products required to complete your real-time PCR experiments.
Pricing for PrimePCR real-time PCR assays is based on assay type, the number of reactions, and plate configuration for 96- and 384-well plates.
Bulk discounts are shown below; download the pricing flyer for full details.
Real-Time PCR Assays and Controls
5 or more assays
Pathway and Collection Panels
20 or more plates
Custom PlatesCustom 96-well & 384-well paltes with 1–96 unique genes
Custom 384-well plates with 97 or more unique genes
30 or more plates
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