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Related Experiment Video

Updated: Dec 24, 2025

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method
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MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as A Novel Detection and Quantification Method

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Homogeneous multiplexed digital detection of microRNA with ligation-rolling circle amplification.

Zhian Hu1, Fujian Xu1, Gongwei Sun1

  • 1Department of Chemistry, Tsinghua University, Beijing 100084, China. sczhang@mail.tsinghua.edu.cn.

Chemical Communications (Cambridge, England)
|April 15, 2020
PubMed
Summary

This study introduces a simple method for detecting multiple microRNAs simultaneously using fluorescence flow cytometry. The technique converts microRNAs into nanoflower balls for direct counting, enabling efficient digital detection.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • MicroRNA (miRNA) detection is crucial for understanding biological processes and disease diagnostics.
  • Existing multiplexed miRNA detection methods can be complex and time-consuming.
  • There is a need for simple, efficient, and accurate digital miRNA detection strategies.

Purpose of the Study:

  • To develop a simple multiplexed digital microRNA detection strategy.
  • To enable simultaneous detection and direct counting of multiple microRNAs.
  • To utilize fluorescence flow cytometry for high-throughput analysis.

Main Methods:

  • Proposed a novel strategy for multiplexed digital microRNA detection.
  • Employed isothermal ligation-rolling circle amplification to convert miRNAs.
  • Generated series of nanoflower balls (NFBs) as digital reporters.
  • Utilized fluorescence flow cytometry for direct counting of NFBs.

Main Results:

  • Successfully achieved simultaneous detection of multiplexed microRNAs.
  • Demonstrated the conversion of miRNAs into countable nanoflower balls.
  • Showcased direct counting of NFBs by fluorescence flow cytometry.
  • Established a simple and effective digital miRNA detection approach.

Conclusions:

  • The proposed strategy offers a simple and effective method for multiplexed digital microRNA detection.
  • Isothermal ligation-rolling circle amplification coupled with flow cytometry provides a powerful tool for miRNA quantification.
  • This approach has potential applications in diagnostics and biological research requiring high-throughput miRNA analysis.