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microRNA PCR Services

We offer a complete microRNA profiling service using our highly sensitive and specific microRNA PCR system - performed by those who know the system best. Our expert scientists perform your microRNA profiling in our state-of-the-art automated laboratories with rigorous quality control and maximum reproducibility tailored to your research needs and budget.

Exiqon, now a QIAGEN company


Exiqon and QIAGEN have joined forces to bring you an enriched product and service offering that builds on the expertise and product synergies of the two companies.

Effective 1 October 2017, Exiqon products and services are fully integrated with QIAGEN, and available for purchase only through QIAGEN.

Meet the new face of Exiqon Services.
Exiqon microRNA PCR Service at a glance:
  • Based on our highly specific and sensitive miRCURY LNA™ Universal RT microRNA PCR system
  • Covers everything from the initial consultation to the final report including all raw data and detailed data analysis
  • Consultation with microRNA PCR experts throughout the process
  • System flexibility and sensitivity ensures cost-efficient experimental setup
  • Experiments performed by expert scientists in state-of-the-art laboratories
  • Rigorous quality control in all steps
  • Normalization procedure optimized for each individual project

A complete microRNA PCR Service

Exiqon's microRNA PCR experts will ensure that you get the best service throughout the projects, from an initial consultation and tailored experimental setup to the data analysis and scientific follow-up. Data and results from your microRNA profiling project will be delivered in an easy-to-read report with publication grade illustrations including an Excel file with all the raw data.

The standard workflow of an Exiqon microRNA PCR Services project is summarized in Figure 1. Details of each step in the process are described in the following tab. Please contact us for further details.
Figure 1 LNA™ improves microRNA PCR performance.
Workflow of a microRNA PCR Services project (Click to learn more)

1. Consultation and experimental design

When you engage in a microRNA PCR Service project with Exiqon, you are assured direct communication with the scientists performing your experiments throughout the duration of the project. Each project begins with a free consultation with a microRNA expression profiling expert. Together, we design an experimental setup that best satisfies your research needs and budget. Following this we will complete a detailed sample-submission form making sure that all experimental details and subsequent analyses are clearly defined and understood by both parties.

2. RNA sample

High quality samples are important for accurate microRNA quantification. At the initial consultation, we offer recommendations on suitable RNA extraction and clean-up methods. Alternatively, you can take advantage of our expertise and submit your samples to our RNA Isolation Service. Due to the universal RT reaction and the sensitivity of our miRCURY LNA™ microRNA PCR system, we can perform analysis with minute RNA input. We normally use 40ng total RNA for full genome profiling and RNA corresponding to just 16μl plasma for profiling with our Serum/Plasma Focus microRNA PCR Panels.

3. RNA sample quality control (QC)

After receiving your RNA samples our specialists will assess the integrity of each sample and determine its concentration. This quality control is not relevant for sample types with no ribosomal RNA such as RNA extracted from blood serum and plasma. In addition, we offer PCR-based QC tests to assess the performance of samples prior to microRNA qPCR profiling. Samples are tested in PCR assays for the amplification of selected microRNAs and spike-ins. Amplification levels are tested to be within known ranges and compared between samples to identify potential outliers. Two tests are offered: basic and extended (See details below).

qPCR based quality control of samples:

RNA PCR QC – Basic:
  • Quantification of four pre-selected microRNAs and a synthetic spike-in.
  • A single RNA input is used.
  • Useful for identification of outlier samples that should be omitted or replaced in the study.
We recommend that this test is performed on all samples.

RNA PCR QC – Extended:
  • Quantification of four pre-selected microRNAs and a synthetic spike-in
  • Three different RNA input amounts are tested
  • Useful for determination of optimal RNA input
  • Useful for detection of inhibitors of the RT and PCR reactions
      This test is recommended for samples types where there is a risk of presence of PCR inhibitors and for sample types we have little experience with. We recommend testing of one sample per experimental group.


    4. microRNA qPCR

    All PCR reactions are performed using our miRCURY LNA™ Universal RT microRNA PCR products which give superior experimental flexibility and allow you to identify and focus on the microRNAs that carry information: full genome profiling with miRNome Panels, Focus Serum/Plasma and Cancer Panels and custom-defined Pick-&-Mix Panels.

    Our state-of-the-art laboratories use high throughput robotic pipetting stations that ensure superior reproducibility. Risk of template contamination is minimized by performing cDNA synthesis and PCR reactions in separate locations. All projects include negative control samples to determine background levels.

    Did you know?

    Our miRCURY LNA™ Universal RT microRNA PCR system is ideally suited for the detection of microRNA in difficult samples such as FFPE and biofluids including serum and plasma.


    5. PCR quality assessment

    Due to their small size microRNA are extremely challenging PCR targets. We have therefore developed a unique automated QC system that allows for careful analysis of the quality of each individual PCR reaction prior to data analysis. Melt curves are inspected, amplification efficiencies are calculated and quantification cycle(Cq) values are compared to background levels in the negative control samples. Based on these analysis reactions are flagged and removed from the data set if they show:
    • Several melting points or have melting points that are not within assay specifications
    • Amplifications with efficiencies outside our accepted range
    • Amplifications with Cq values within a threshold range of background signal

    6. Normalization and data analysis

    Before data analysis, we make sure your data are normalized to correct for potential overall differences between samples. The method of normalization is optimized for each individual project using sophisticated software packages. In case of biological replicates average ΔCq values are calculated and ΔΔCq values are determined based on the biological grouping of samples.

    Data analysis appropriate for the experiment at hand is performed, e.g., Principle Component Analysis (PCA) and heat maps based on unsupervised clustering. We check if samples group according to biology and look for any signs of experimental bias in the data set. Finally, statistical analysis of customer defined group comparisons is performed.

    In connection with large service projects we also offer an extensive customized bioinformatics analysis.

    7. Report and final consultation

    An easy-to-read summary report is provided containing a description of the project, assessments of sample and data quality and an overview of the results of the data analysis with publication-grade illustrations. An extensive Excel file with all raw data is also included. The final report is delivered as a link to our secure webserver. See the tab “PCR Report” for further details on the content of the final report.

    Scientific follow-up can be organized in case you have questions to your final report or wish to discuss the significance of your results further.
    Figure 1

    Biomarker discovery in colorectal cancer (CRC)

    Exiqon has extensive knowledge of microRNA profiling in a wide range of samples and has been involved in numerous biomarker discovery projects.

    We are currently working on an advanced and promising program on early detection of colorectal cancer (CRC) by microRNA analysis of patient blood plasma using our microRNA qPCR platform. Experimental details on this study are presented below and in Figure 1.
     

    *See experimental setup in figure 1.

    In addition to our extensive knowledge of biomarkers, we are experts on our own qPCR system. We are therefore uniquely positioned to assist you in all phases of biomarker discovery projects – from experimental design guidance to advanced bioinformatics. An overview of the general biomarker discovery project setup is presented below.
     

    Biomarker discovery workflow using Exiqon’s microRNA PCR system. (Click to learn more)

    Figure 1
    Download our new conference poster regarding our study on early detection of CRC.

    The result of your microRNA PCR study is presented in an easy-to-read summary report

    The report contains description of the project, assessments of sample and data quality and an overview of the results of the data analysis with publication-grade illustrations.

    An extensive Excel file with all raw data is also included. The final report is delivered as link to our secure webserver. Scientific follow-up can be organized in case you have questions to your final report or wish to discuss the significance of your results further.

    The microRNA PCR Profiling report consists of:


    An RNA sample quality control (QC) report

    An easy-to-read summary report with publication-grade illustrations
    • Description of the project and workflow
    • QA/QC of sample and data
    • Plots showing the effect of normalization
    • PCA plots and heat maps
    • Heat maps based on unsupervised and supervised clustering
    • Results of statistical analysis of customer defined group comparisons.
    • Summary of results.
    • If relevant, data is compared to Exiqon’s in-house microRNA expression database
    A material and methods section
    • Ready to use for publication purposes.
    Project summary Excel report with publication-grade illustrations
    • Raw and normalized Cq values
    • Statistical analysis and heat maps according to customer specifications
    Figure 1
    Dr. Wojciech Fendler

    microRNAs as disease biomarkers


    "Excellent technical and scientific feedback allows researchers from different fields to incorporate miRNA experiments into their study protocols."

    Dr. Wojciech Fendler works in the Department of Pediatrics, Oncology, Hematology and Diabetology at the Medical University of Lodz, Poland. He is interested in the application of microRNAs as biomarkers in disease and has performed microRNA profiling using Exiqon Services.

    Read full story...

    Dr. Anja Bye

    Can microRNAs tell you how fit you are?


    "It was essential to us that the RNA-isolation service was available and that we could get our results within a short time frame. Finally, it was important that the service provider had a good reputation."

    Our research focuses on identifying the key cellular and molecular mechanisms underlying the beneficial effects of physical exercise on the heart, arteries and skeletal muscle in the context of disease prevention and management through experimental, clinical and epidemiological studies. Identifying the cellular and molecular mechanisms associated with aerobic fitness is important, because it may help us develop new and better methods to prevent and treat cardiovascular disease.

    Read full story...

    Dr. Wendy Alderton

    Determining the quality of biobank serum for microRNA analysis


    "Exiqon staff were extremely helpful in helping to design the study, analyze and interpret the results for us using all their past experience in this area."

    Abcodia is engaged in the validation and discovery of biomarkers for the early detection and screening of disease and has exclusive commercial license to a large prospective serum biobank. They have recently completed a successful pilot study on microRNA detection which confirms the quality of the UKCTOCS serum for microRNA discovery and validation projects.

    Read full story...

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