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Related Concept Videos

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Ribosome Profiling

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Related Experiment Video

Updated: May 28, 2026

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
07:27

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs

Published on: August 3, 2011

Label-free high-throughput microRNA expression profiling from total RNA.

Demin Duan1, Ke-xiao Zheng, Ye Shen

  • 1Suzhou Institute of Nano-tech and Nano-bionics, Chinese Academy of Sciences, 398 Ruoshui Road, Suzhou, 215123, China.

Nucleic Acids Research
|October 7, 2011
PubMed
Summary

This study introduces a label-free method for microRNA (miRNA) detection using a novel hybridization-triggered fluorescence strategy. The Stacking-Hybridized Universal Tag (SHUT) assay simplifies miRNA profiling from total RNA, enhancing accuracy and efficiency.

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Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays
10:58

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR (qPCR) Arrays

Published on: December 3, 2010

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • MicroRNAs (miRNAs) are crucial biological regulators and potential disease biomarkers.
  • Current miRNA detection methods often require labeling, which is labor-intensive, time-consuming, and prone to bias.
  • Existing assays struggle to differentiate between mature miRNAs and precursors (pre-miRNAs) when using total RNA.

Purpose of the Study:

  • To develop a label-free, high-throughput method for miRNA expression profiling.
  • To overcome the limitations of labeling-based miRNA detection techniques.
  • To address the challenge of cross-hybridization with pre-miRNAs in total RNA samples.

Main Methods:

  • A hybridization-triggered fluorescence strategy was employed for microarray-based analysis.
  • The Stacking-Hybridized Universal Tag (SHUT) assay utilizes a fluorophore-linked oligonucleotide Universal Tag.
  • Direct capture of the Universal Tag by target-bound probes occurs via base-stacking interactions.

Main Results:

  • The SHUT assay enables label-free, high-throughput miRNA expression profiling directly from total RNA.
  • The assay successfully detected miRNA expression using as little as 100 ng of human tissue total RNA.
  • Demonstrated high specificity for homogenous miRNAs and superb discrimination against single-base mismatches.
  • Pre-miRNAs produced negligible signals, confirming the assay's ability to directly use total RNA.

Conclusions:

  • The SHUT assay offers a simplified, efficient, and accurate approach for miRNA profiling.
  • This label-free method eliminates the need for RNA labeling, reducing bias and labor.
  • The assay's specificity and ability to use total RNA advance miRNA detection for research and diagnostics.