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Related Experiment Video

Updated: Dec 10, 2025

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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DSN/TdT recycling digestion based cyclic amplification strategy for microRNA assay.

Jing-Lin He1, Ting-Ting Mei1, Ling Tang1

  • 1Hunan Provincial Key Laboratory of Materials Protection for Electric Power and Transportation, Hunan Provincial Key Laboratory of Cytochemistry, School of Chemistry and Food Engineering, Changsha University of Science and Technology, Changsha, 410114, PR China.

Talanta
|September 5, 2020
PubMed
Summary

This study introduces a novel fluorescence signal amplification strategy for detecting microRNAs (miRNAs) using DSN/TdT recycling digestion. This method offers sensitive and specific miRNA detection, crucial for early cancer diagnosis and biomarker analysis.

Keywords:
Duplex-specific nucleaseMicroRNARecycling digestionTerminal deoxynucleotidyl transferase

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Sensitive and specific detection of microRNAs (miRNAs) is critical for early cancer diagnosis.
  • Existing methods may lack sensitivity or require complex designs.

Purpose of the Study:

  • To develop a simple and sensitive fluorescence signal amplification strategy for miRNA detection.
  • To enable efficient and sequence-independent miRNA analysis.

Main Methods:

  • Utilized a DSN/TdT recycling digestion and extension mechanism for signal amplification.
  • DSN enzyme initiates digestion on miRNA heteroduplexes, enabling miRNA recycling.
  • TdT enzyme synthesizes polydeoxyguanylic tails, leading to dsDNA formation and further DSN/TdT cycles.

Main Results:

  • Achieved massive fluorophore separation and significant signal amplification.
  • Demonstrated sequence-independent reactions for both DSN digestion and TdT extension.
  • Successfully analyzed miRNA samples from MCF-7 cell lysates and Cu (II) ion samples.

Conclusions:

  • The developed strategy provides a promising analytical platform for DNA nicking-related studies.
  • This method holds potential for tumor biomarker measurement in clinical diagnostics.
  • The approach is simple, sensitive, and applicable to real-world sample analysis.