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Related Experiment Video

Updated: Feb 21, 2026

High Throughput MicroRNA Profiling: Optimized Multiplex qRT-PCR at Nanoliter Scale on the Fluidigm Dynamic ArrayTM IFCs
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Using FirePlex™ Particle Technology for Multiplex MicroRNA Profiling Without RNA Purification.

Michael R Tackett1, Izzuddin Diwan2

  • 1Abcam Inc., One Kendall Square Suite, B2304, Cambridge, MA, 02139, USA. Michael.Tackett@abcam.com.

Methods in Molecular Biology (Clifton, N.J.)
|October 8, 2017
PubMed
Summary

This study introduces a novel multiplex assay for profiling circulating microRNAs (miRNAs) directly from plasma or serum without RNA purification. This method enhances accuracy and simplifies analysis for biomarker discovery.

Keywords:
High-throughputHybridizationMultiplexPolymerase chain reactionmicroRNA

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genomics

Background:

  • MicroRNA (miRNA) profiling accuracy is critical for biomarker discovery.
  • Traditional methods often involve RNA purification, introducing potential bias and inaccuracies.
  • Minimizing sample processing steps is key to enhancing miRNA assay reliability.

Purpose of the Study:

  • To present a novel multiplex circulating miRNA assay for direct profiling from biofluids.
  • To demonstrate the assay's ability to profile up to 65 miRNAs simultaneously.
  • To eliminate the need for RNA purification, thereby reducing bias.

Main Methods:

  • Utilizing FirePlex™ hydrogel particles for multiplex miRNA capture.
  • Performing direct analysis from plasma or serum samples (including heparin plasma).
  • Employing conventional flow cytometry and user-friendly software for data acquisition and analysis.

Main Results:

  • Achieving picomolar sensitivity and high specificity in miRNA capture.
  • Enabling profiling of up to 65 miRNAs in a single reaction well.
  • Demonstrating PCR-level sensitivity from small volumes of crude biofluid (10 μL) or purified RNA (<100 pg).

Conclusions:

  • The novel assay provides an accurate and efficient method for circulating miRNA profiling.
  • Direct analysis from biofluids simplifies the workflow and minimizes experimental bias.
  • This approach facilitates rapid and reliable miRNA biomarker discovery.