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Related Experiment Video

Updated: Jul 24, 2025

Quantification of Circular RNAs Using Digital Droplet PCR
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Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

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Single-Cell Analysis of circRNA Using ddPCR.

Jiayi Peng1, Feng Li1, Xiangdong Xu1

  • 1Division of Oral and Systemic Health Sciences, School of Dentistry, University of California, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 10, 2023
PubMed
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Droplet digital PCR (ddPCR) offers sensitive quantification of nucleic acids. This method is applied here to measure circular RNAs (circRNAs) in single pancreatic cancer cells.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genomics

Background:

  • Circular RNAs (circRNAs) are increasingly recognized for their roles in cancer development and progression.
  • CircRNAs hold potential as diagnostic and prognostic biomarkers in oncology.
  • Novel methods are needed for precise quantification of circRNAs, especially in single cells.

Purpose of the Study:

  • To describe the detailed procedures for quantifying circular RNAs (circRNAs) in individual pancreatic cancer cells.
  • To demonstrate the application of Droplet digital PCR (ddPCR) for sensitive circRNA analysis.
  • To establish a methodology for single-cell circRNA profiling in cancer research.

Main Methods:

  • Utilizing Droplet digital PCR (ddPCR) technology for nucleic acid quantification.
Keywords:
Droplet digital PCRPancreatic cancer cellsSingle-cell analysiscircRNA

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Last Updated: Jul 24, 2025

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  • Fractionating samples into approximately 20,000 individual nanoliter-sized droplets for PCR amplification.
  • Employing automated droplet readers to analyze fluorescence signals for accurate quantitation.

    • Main Results:

    • ddPCR provides highly sensitive and accurate quantification of nucleic acid molecules, even at low copy numbers.
    • The described methodology allows for the precise measurement of circRNA levels within single pancreatic cancer cells.
    • This approach enables detailed analysis of circRNA heterogeneity at the single-cell level.

    Conclusions:

    • Droplet digital PCR is a powerful tool for the sensitive and accurate quantification of circRNAs.
    • The described single-cell ddPCR method facilitates the study of circRNA expression in pancreatic cancer.
    • This technique can advance the understanding of circRNA functions and their utility as cancer biomarkers.