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...

PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

You might also read

Related Articles

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

Sort by

Same author

Interrelationships Among Physical Fitness, General Motor Coordination, and Soccer-Specific Technical Skills in Youth Soccer Players.

Sports (Basel, Switzerland)·2026

Same author

Clinical and vaginal microbiota effects of oral Lactobacillus crispatus M247 combined with vaginal laser therapy in menopausal women with atrophic vulvovaginitis: a prospective, randomized and controlled study.

Lasers in medical science·2026

Same author

Ketogenic Diets and Low Carb Diets: The Effects on the Intestinal Microbiota. A Narrative Review.

Nutrition reviews·2026

Same author

Extracellular vesicles released by uterine fibroid cells: modulation by omega-3 fatty acids and impact on recipient myometrial cells.

F&S science·2026

Same author

Role of <i>Bifidobacterium breve</i> PRL2020 in reducing symptoms of antibiotic-associated diarrhea in children.

Frontiers in pediatrics·2026

Same author

Effects of combined Mediterranean diet and physical activity intervention on the gut microbiome and disease progression in individuals with Parkinson's disease: study protocol for a multicenter, randomized controlled pilot study (PRIME study).

Frontiers in aging neuroscience·2026

Search research articles

Related Experiment Video

Updated: Jun 13, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Shape based kinetic outlier detection in real-time PCR.

Davide Sisti¹, Michele Guescini, Marco B L Rocchi

¹Dipartimento DiSUAN, Sezione di Biomatematica, Università degli Studi di Urbino Carlo Bo, Campus Scientifico Sogesta; Località Crocicchia - 61029 Urbino, Italy. davide.sisti@uniurb.it

BMC Bioinformatics

|April 14, 2010

Summary

A new strategy, Sample outlier detection (SOD), analyzes amplification curve shapes to identify outlier samples in real-time PCR. SOD is more sensitive than current methods for detecting outliers caused by inhibitors, improving quantification reliability.

More Related Videos

The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform

The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform

Published on: September 27, 2016

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Related Experiment Videos

Last Updated: Jun 13, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform

The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform

Published on: September 27, 2016

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Published on: June 8, 2019

Area of Science:

Molecular Biology
Biotechnology
Genomics

Background:

Real-time PCR is the preferred method for nucleic acid quantification.
Accurate quantification relies on the assumption of similar PCR efficiency between samples.
Biological sample components can inhibit PCR kinetics, affecting quantification accuracy.

Purpose of the Study:

To introduce a novel strategy, Sample outlier detection (SOD), for identifying outlier samples in real-time PCR.
To assess SOD's ability to detect outliers caused by PCR inhibitors.
To compare SOD's performance against existing methods.

Main Methods:

Richards function was used to parameterize amplification curves from fluorescence readings.
Amplification curve parameters (plateau, slope, inflection point) were analyzed.
SOD compared parameters of unknown samples to standards to detect outliers.

Main Results:

Amplification parameters did not correlate with input DNA, providing a unique fingerprint for each curve.
SOD effectively identified outlier profiles affected by inhibitors like tannic acid, IgG, and quercitin.
SOD demonstrated higher sensitivity than the current KOD method, with comparable specificity.

Conclusions:

Outlier detection in real-time PCR can be based on amplification curve shape, not solely PCR efficiency.
SOD offers improved real-time PCR analysis by reducing data variance.
This leads to more reliable nucleic acid quantification.