circRNA qPCR Service

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circRNA qPCR Service

IntegrateRNA provides a comprehensive set of circRNA qPCR (quantitative PCR) services, including divergent primer design, RNA isolation/QC, RNase R treatment, cDNA synthesis, qPCR and data analysis. We are confident to manage the entire circRNA qPCR services workflow and deliver reliable data in a timely and cost-effective manner.

Circular RNA (circRNA), the 3' and 5' ends of which are covalently linked, constitutes a class of RNA recently discovered to be widespread and abundant. circRNAs are generally formed by alternative splicing of pre-mRNA in which an upstream splice acceptor is joined to a downstream splice donor in a process known as 'backsplicing'. Recent studies show that circRNA is a novel potential biomarker for disease diagnosis, also functions as miRNA/RBP sponge to play regulatory role in disease development and progression.

Although RNA-seq analysis and microarray-based expression profiling analysis provide essential information regarding circRNA junctions and annotation, it’s necessary to perform biochemical experiment to validate high-throughput results. Among the most basic tools to validate a circRNA is RT-qPCR (reverse transcription followed by quantitative polymerase chain reaction).

Workflow of circRNA qPCR Service

  • Divergent primer design
  • The directions of the primers face toward each other for the circular RNA sequence but away from each other with the linear RNA. This prevents the amplification of species that do not contain the backsplice junction, such as mRNA or genomic DNA.

    Divergent primers designed for circRNA junction sitesFigure 1. Divergent primers designed for circRNA junction sites

  • Total RNA isolation and quality control
  • Degradation of linear RNA by digestion with RNase R and cDNA synthesis
  • PCR and circRNA sequencing
  • qPCR analysis of circRNA

Benefits

  • Our scientists and lab personnel are experts in TaqMan and SYBR Green chemistries.
  • We can identify suitable endogenous controls for your experiments.
  • Our complete qPCR service workflow includes RNA QC, application-specific sample processing, divergent primer design, qPCR, and data analysis according to your specific needs.

Deliverable

  • Divergent primers (If required)
  • Detailed experiment report
  • Raw data, amplification curve and melting curve of qPCR

As a leading biotechnology company, IntegrateRNA has extensive experience with all types of RNA discovery, analysis, and related bioinformatics services, and is dedicated to providing you the most comprehensive and professional RNA services. Our experienced team of experts performs quality management and follows each process to ensure confident and fair results. If you have any questions or needs, please feel free to contact us.

References:

  1. Panda A C. et al. Detection and Analysis of Circular RNAs by RT-PCR. Bio-protocol, 2018, 8(6).
  2. Barrett S P. et al. Circular RNAs: Analysis, expression and potential functions. Development, 2016, 143(11):1838-1847.
For research use only. Not intended for any clinical use.
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