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RT-qPCR Services
Real Time Quantitative PCR (RT-qPCR) is a powerful tool for analyzing gene expression. By quantitating gene transcript, RT-qPCR reveals trends in cellular response to disease, drugs, and signaling processes.
Ichor’s RT-qPCR services combine state-of-the-art instrumentation with expert execution to achieve accurate and reliable results. Our Applied Biosystems QuantStudio 3 and QuantStudio 5 platforms offer incredible experimental flexibility, without compromising sensitivity.
Several Reporter Options
Our platform can analyze reactions containing Taq-Man probes or SYBR intercalating agents, providing maximum experimental flexibility.
Broad Dye Compatibility
Multiple filter sets provide several options for probe labeling, and enable multiplex analysis.
Sample Preparation Support
Full RT-qPCR service can include sample preparation and processing, or can utilize customer-provided templates.
Multiplex Analysis
Interrogating multiple analytes from a single sample yields more accurate results with less time and materials.
Single Copy Detection
Our RT-qPCR platform provides unsurpassed accuracy and a broad dynamic range, allowing detection of single copy templates.
Comprehensive RT-qPCR Support
Ichor offers end-to-end RT-qPCR service, and can provide as much support as is needed to ensure the success of your project.
Meticulous Optimization Yields Better Multiplex Results
For RT-qPCR reactions containing multiple primer pairs, primer design must account for Tm compatibility while avoiding primer dimer occurrence. To ensure the best performance, we acquire pre-validated primers from trusted vendors. Once received, each primer pair is tested in gradient runs individually, and then in combination, to optimize amplification parameters. Agarose gel electrophoresis is performed during initial optimization to validate appropriate product formation, and melt curve analysis is performed at every run to ensure that fluorescence signal is attributed to appropriate amplicons.
Amplification plot and standard curve of 1047bp DNA template. (A) Amplification plot with different shade of blue colors represented different concentrations of template which were illustrated in the figure. (B) Standard curve of Ct value versus template concentration.
Transcript Response to Drugs and Disease
Featured Capabilities / Case Studies
Gene Expression Profiling
Characterize Dynamic Cell Behaviors
Gene regulation underlies cellular response to changing conditions. Ichor’s RT-qPCR services offer streamlined gene expression analysis to characterize cell response. Custom primer and probe design are integrated into our established protocols for reverse transcription and RT-qPCR analysis to provide accurate and reliable expression profiles.
Here, cells in culture were treated and analyzed longitudinally for expression of several mRNA transcripts. CT values for each transcript of interest were normalized to those of a housekeeping gene and were graphed as the fold change in expression compared to 0 hr. using the ΔΔCT method.
Relative gene expression profiling can be used to analyze transcriptional changes in primary or immortalized cells in culture, or can be applied to tissue and blood isolated from human or animal studies.
Absolute Quantitation
Template Copy Number Determination
RT-qPCR can be used for absolute copy number quantitation of template DNA. By producing a purified template of known concentration, a standard curve can be established to relate template copy number to CT value. Using this curve, templates of unknown concentration are subjected to identical reaction conditions, and CT values are used to back-calculate initial template DNA quantities.
Here, a standard curve was generated by subjecting serial dilutions of template DNA to RT-qPCR amplification and analysis. The resulting standard curve was plotted on a log scale and showed a good linear regression fit, allowing for absolute quantitation of DNA template from two unknown samples.
Gene Induction in Disease Models
Transcriptional Changes Reflect Cellular Response to Small Molecules
Here, U87-MG cells/animals were treated/inoculated with single-dose of 10 uM Alendronate. Cells/tissues were harvested at designated time, and analyzed by RT-qPCR to characterize the change in expression level of CCL20 genes relative to ACTB1 house-keeping gene.
Similar assessments can also measure the cellular response to disease interventions. Ichor’s RT-qPCR services offer customizable transcript profiling workflows to ensure that changes in response to disease or drugs are captured and characterized.
Frequently Asked Questions
Find out more about our customizable RT-qPCR service offerings.