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Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

Updated: May 30, 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

Ultrasensitive multiplexed microRNA quantification on encoded gel microparticles using rolling circle amplification.

Stephen C Chapin1, Patrick S Doyle

  • 1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, United States.

Analytical Chemistry
|August 5, 2011
PubMed
Summary

This study presents a novel method for precisely quantifying microRNAs (miRNAs) in complex biological samples. The technique achieves high sensitivity and enables direct detection in unprocessed serum, paving the way for rapid diagnostics.

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Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
10:28

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Related Experiment Videos

Last Updated: May 30, 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

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
10:28

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Accurate quantification of multiple biomolecules in complex samples is challenging.
  • Existing methods often require extensive sample preparation and can suffer from amplification bias.
  • There is a need for sensitive, direct detection platforms for biomarkers like microRNAs (miRNAs).

Purpose of the Study:

  • To develop a flexible platform for multiplexed quantification of microRNAs (miRNAs).
  • To achieve subfemtomolar sensitivity and single-molecule resolution in biomolecule analysis.
  • To enable direct detection of miRNAs in unprocessed biological samples like human serum.

Main Methods:

  • Utilized encoded hydrogel microparticles for capturing miRNA targets.
  • Employed rolling circle amplification (RCA) of a universal adapter sequence for signal amplification.
  • Integrated multiple fluorescent reporters for enhanced detection and eliminated amplification bias.
  • Leveraged gel particle resistance to fouling for direct sample analysis.

Main Results:

  • Achieved subfemtomolar sensitivity in multiplexed miRNA quantification.
  • Demonstrated single-molecule reporting resolution.
  • Successfully detected miRNAs directly in small quantities of unprocessed human serum.
  • Eliminated the need for RNA extraction or target-amplification steps.

Conclusions:

  • The developed hydrogel microparticle platform offers a sensitive and versatile solution for biomolecule analysis.
  • The RCA-based approach provides high sensitivity and eliminates amplification bias.
  • Direct detection in unprocessed serum has significant implications for developing rapid, noninvasive diagnostic assays.