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Related Concept Videos

Real Time RT-PCR02:57

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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
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Quantification of Circular RNAs Using Digital Droplet PCR
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Profiling of circRNAs using an enzyme-free digital counting method.

Lasse Sommer Kristensen1

  • 1Department of Biomedicine, Aarhus University, Denmark.

Methods (San Diego, Calif.)
|February 14, 2021
PubMed
Summary

Circular RNAs (circRNAs) are promising disease biomarkers, but their detection is challenging. An enzyme-free NanoString nCounter method offers accurate quantification of these circular RNA molecules.

Keywords:
Accurate quantificationBiomarkersCancerCircular RNANanoString nCounter

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Identification of Circular RNAs using RNA Sequencing
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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • Circular RNAs (circRNAs) are increasingly recognized as significant biomarkers for various human diseases, particularly cancer.
  • The unique circular structure of circRNAs presents challenges for accurate detection and quantification.
  • Enzymatic methods, like those using reverse transcriptases, can compromise circRNA quantification accuracy and reproducibility.

Purpose of the Study:

  • To review the advantages of the NanoString nCounter technology for circRNA quantification.
  • To discuss methodological considerations, potential pitfalls, and disadvantages associated with this enzyme-free digital method.

Main Methods:

  • Review of existing literature on circRNA detection and quantification.
  • Focus on the application and adaptation of the NanoString nCounter platform for circRNA analysis.
  • Discussion of enzyme-free digital quantification principles.

Main Results:

  • The NanoString nCounter platform provides an enzyme-free approach for digital circRNA quantification.
  • This method addresses limitations associated with traditional enzymatic detection.
  • Methodological considerations and potential challenges for accurate circRNA measurement are highlighted.

Conclusions:

  • The NanoString nCounter technology presents a valuable tool for accurate circRNA biomarker quantification.
  • Understanding the method's advantages and limitations is crucial for reliable biomarker discovery.
  • Enzyme-free digital detection offers a promising avenue for advancing circRNA-based diagnostics.