Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Structure elucidation of an exopolysaccharide produced by Sphingomonas Sanxanigenens NXO2.

Food chemistry·2025

Same author

Droplet-based Cytotoxicity Assay to Assess Chimeric Antigen Receptor T cells at the Single-cell Level.

Journal of visualized experiments : JoVE·2025

Same author

Efficacy and safety profile of a subtilisin protease produced by fermentation in <i>Bacillus licheniformis</i> to be used as a feed additive.

Heliyon·2022

Same author

Safety and efficacy profile of a phytase produced by fermentation and used as a feed additive.

Heliyon·2021

Same author

Peer preferences and characteristics of same-group and cross-group social interactions among autistic and non-autistic adolescents.

Autism : the international journal of research and practice·2021

Same author

Verification of CRISPR editing and finding transgenic inserts by Xdrop indirect sequence capture followed by short- and long-read sequencing.

Methods (San Diego, Calif.)·2021

Search research articles

Related Experiment Video

Updated: Feb 25, 2026

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

Published on: January 22, 2014

Quantitative RT-PCR for MicroRNAs in Biofluids.

Michael Thorsen¹, Thorarinn Blondal¹, Peter Mouritzen²

¹Exiqon A/S, Skelstedet 16, Vedbæk, DK-2950, Denmark.

Methods in Molecular Biology (Clifton, N.J.)

|July 28, 2017

Summary

This summary is machine-generated.

Researchers developed improved methods for detecting microRNAs in biofluids using quantitative reverse transcription polymerase chain reaction (RT-PCR). These protocols address challenges like low RNA levels and inhibitors, enhancing diagnostic biomarker potential.

Keywords:

Biofluids Expression analysis RNA isolation RT-qPCR microRNA

More Related Videos

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

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

Published on: April 14, 2015

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Published on: February 12, 2018

Related Experiment Videos

Last Updated: Feb 25, 2026

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

Cerebrospinal Fluid MicroRNA Profiling Using Quantitative Real Time PCR

Published on: January 22, 2014

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

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

Published on: April 14, 2015

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Absolute Quantification of Plasma MicroRNA Levels in Cynomolgus Monkeys, Using Quantitative Real-time Reverse Transcription PCR

Published on: February 12, 2018

Area of Science:

Biomarker Discovery
Molecular Diagnostics
Genomics

Background:

MicroRNAs (miRNAs) in biofluids show potential as minimally invasive diagnostic biomarkers.
Quantitative reverse transcription polymerase chain reaction (RT-PCR) is a sensitive method for miRNA detection.
Challenges in biofluid miRNA quantification include low RNA yield, inhibitors, and preanalytical variability.

Purpose of the Study:

To describe optimized procedures for sensitive and accurate miRNA detection in biofluids.
To address challenges associated with miRNA quantification in complex biological samples.
To establish robust protocols for reliable miRNA biomarker development.

Main Methods:

Development of highly sensitive miRNA detection assays.
Optimization of sample handling and preparation protocols for biofluids.
Implementation of extensive quality control (QC) measures throughout the workflow.

Main Results:

Established procedures that overcome common limitations in biofluid miRNA analysis.
Demonstrated the feasibility of accurate miRNA quantification despite low RNA levels and inhibitors.
Ensured reliability and reproducibility of miRNA expression measurements.

Conclusions:

Optimized protocols enhance the utility of biofluid miRNAs as diagnostic biomarkers.
The described methods improve the sensitivity and accuracy of miRNA quantification.
These advancements facilitate the development of minimally invasive diagnostic tools.