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Real Time RT-PCR

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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Updated: Feb 21, 2026

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

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

Published on: June 26, 2016

Microfluidics-based digital quantitative PCR for single-cell small RNA quantification.

Tian Yu¹, Chong Tang¹, Ying Zhang¹

¹Department of Physiology and Cell Biology, University of Nevada, Reno School of Medicine, Reno, Nevada, USA.

Biology of Reproduction

|October 13, 2017

Summary

This summary is machine-generated.

A new microfluidics-based digital quantitative PCR (mdqPCR) method enables sensitive small RNA analysis in single cells. This technique revealed unique microRNA (miRNA) profiles in individual sperm cells.

Keywords:

RNA quantification digital PCR fertility germ cells small RNA sperm

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Quantification of Circular RNAs Using Digital Droplet PCR

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Last Updated: Feb 21, 2026

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

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Quantification of Circular RNAs Using Digital Droplet PCR

Quantification of Circular RNAs Using Digital Droplet PCR

Published on: September 16, 2022

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Quantitative PCR (qPCR) methods face limitations in sensitivity and specificity for single-cell small RNA analysis.
Existing digital quantitative PCR (dqPCR) methods are restricted to miRNA quantification and require proprietary reagents.

Purpose of the Study:

To develop a novel microfluidics-based digital quantitative PCR (mdqPCR) method for high-throughput, sensitive small RNA quantification at the single-cell level.
To assess the applicability of mdqPCR for analyzing diverse small RNA species beyond miRNAs.

Main Methods:

Utilized the Fluidigm BioMark HD system for template partitioning and high-throughput digital quantitative PCR (dqPCR).
Developed a microfluidics-based approach (mdqPCR) for comprehensive small RNA analysis.

Main Results:

The mdqPCR method demonstrated high sensitivity and reproducibility for single-cell small RNA quantification.
Successfully quantified not only miRNAs but also other small RNA species at the single-cell level.
Discovered unique microRNA (miRNA) profiles in individual sperm cells.

Conclusions:

The developed mdqPCR method overcomes limitations of conventional techniques for single-cell small RNA analysis.
mdqPCR offers a versatile and sensitive platform for diverse small RNA species quantification.
This advancement enables novel discoveries, such as the unique miRNA signatures within individual sperm.