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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 29, 2026

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR
07:37

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

Published on: June 26, 2016

Micro-RNA quantification using DNA polymerase and pyrophosphate quantification.

Hsiang-Ping Yu1, Yi-Ling Hsiao, Hung-Yin Pan

  • 1Institute of Biotechnology, National Taipei University of Technology, 1, Section 3, Taipei 106, Taiwan.

Analytical Biochemistry
|September 14, 2011
PubMed
Summary

A new method rapidly quantifies micro-RNA (miRNA) using DNA polymerase and pyrophosphate detection. This simple, inexpensive assay is highly sensitive and broadly applicable for RNA and DNA detection.

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Related Experiment Videos

Last Updated: May 29, 2026

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR
07:37

Circulating MicroRNA Quantification Using DNA-binding Dye Chemistry and Droplet Digital PCR

Published on: June 26, 2016

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing
14:15

Highly Efficient Ligation of Small RNA Molecules for MicroRNA Quantitation by High-Throughput Sequencing

Published on: November 18, 2014

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:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Accurate quantification of micro-RNA (miRNA) is crucial for understanding gene regulation and disease.
  • Existing methods for miRNA detection can be complex, time-consuming, or lack sensitivity.

Purpose of the Study:

  • To develop a rapid, sensitive, and cost-effective method for miRNA quantification.
  • To demonstrate the assay's applicability to purified RNA samples and its broad potential for RNA and DNA detection.

Main Methods:

  • A novel assay utilizing DNA polymerase activity and pyrophosphate quantification.
  • Micro-RNA acts as a primer, and a DNA probe serves as the template for DNA synthesis.
  • Pyrophosphate detection quantifies the extent of DNA synthesis, indicating miRNA presence and quantity.

Main Results:

  • Detection of as little as five femtomoles of synthetic RNA.
  • Quantification of hsa-miR-16 and hsa-miR-21 in SiHa cell RNA samples (0.34 fmol/μg and 0.71 fmol/μg, respectively).
  • Assay completion in under 5 minutes post-RNA purification, unaffected by pre-miRNA.

Conclusions:

  • The developed method offers a simple, inexpensive, and rapid approach for miRNA quantification.
  • The assay's generality allows for wide application in detecting various RNA and DNA targets.
  • This technique holds significant potential for diverse molecular diagnostic and research applications.