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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.

Your data package includes instrument read files, CT values, and calculated fold-expression data. RT-qPCR services and analysis can be customized according to your needs.

Well-designed primers enable successful RT-qPCR experiments. Our reagents are pre-validated to provide superior performance, and reactions are monitored on an ongoing basis to continually verify quality.

For projects requiring absolute quantitation of template copy number, we generate a standard curve to enable the determination of template copy number from RT-qPCR CT value.

For difficult reactions, or to identify appropriate parameters for multiplex experiments, we conduct initial optimization of amplification conditions to ensure reliable assay performance.

Take advantage of our in-house veterinary and cell culture capabilities to streamline your process. We can conduct the experiments, harvest samples, prepare templates, and run RT-qPCR in one integrated workflow.

Meticulous Optimization Yields Better Multiplex Results

Cycle _DNA

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. 

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.

Rapid and Reliable RT-qPCR Results

Our high-quality RT-qPCR services deliver accurate results, with shorter turnaround times than the competition.

Frequently Asked Questions

Find out more about our customizable RT-qPCR service offerings.

Ichor provides whatever processing you need.  We accept cell pellets/harvested tissue, extracted RNA, or cDNA. We can also perform your cell or animal treatments in-house to streamline your workflow.

Your data package includes raw read files (or exported data), along with desired endpoints and analyses, including CT values, and standard curve or fold expression calculations. Custom analyses are always offered.

Yes- we work with trusted vendors for the acquisition of primers and probes. Our suppliers provide pre-validated reagents, and we always confirm performance in-house prior to executing your assay.

When you partner with us, you gain access to our experienced team of scientists, who can perform initial optimization to identify appropriate conditions for cell or animal treatments, and can optimize reactions for downstream RT-qPCR.