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

Estimation of gut microbiome motif associated with active tuberculosis - A case control study.

Bioinformation·2026

Same author

ML-FSID-FIS: A Multi-Level Feature Selection and Fuzzy Inference System for Intrusion Detection in IoMT.

Sensors (Basel, Switzerland)·2026

Same author

Generation of biologically responsive colon-like intestinal tissue patches from human induced pluripotent stem cells using a rapid co-differentiation platform.

Stem cell research & therapy·2026

Same author

FedSMOTE-DP: Privacy-Aware Federated Ensemble Learning for Intrusion Detection in IoMT Networks.

Sensors (Basel, Switzerland)·2026

Same author

A Prospective Analysis of Maternal and Fetal Outcomes in Term Pregnancies Complicated by Oligohydramnios at Bundelkhand Medical College.

Journal of pharmacy & bioallied sciences·2026

Same author

Enhancing Tuberculosis Diagnosis and Treatment Monitoring: Evaluating Ki67 and HLA-DR Biomarkers Via Point-of-Care Testing and Comparative Analysis of Flow Cytometry Versus ELISA in Whole Blood Samples.

Journal of pharmacy & bioallied sciences·2026

Search research articles

Related Experiment Video

Updated: Mar 8, 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

QMC-PCRx: a novel method for rapid mutation detection.

Henry O Ebili^1,2, James Hassall¹, Abutaleb Asiri¹

¹Division of Cancer and Stem Cell, Faculty of Medicine & Health Sciences, University of Nottingham, Nottingham, UK.

Journal of Clinical Pathology

|February 4, 2017

Summary

The novel QMC-PCRx method enables multiplexed mutation screening, significantly reducing workload and costs. This multiplexed analysis using high-resolution melting (HRM) achieves 100% concordance with singleplex testing.

Keywords:

CANCER GENETICS COLORECTAL CANCER PCR

More Related Videos

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Published on: February 5, 2014

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Related Experiment Videos

Last Updated: Mar 8, 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

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Published on: February 5, 2014

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Published on: March 29, 2017

Area of Science:

Molecular biology
Genetics
Biotechnology

Background:

Quick multiplex consensus PCR (QMC-PCR) is a two-stage method for rapid mutation screening in low-quality DNA.
The standard QMC-PCR involves a prediagnostic multiplex (PDM) reaction followed by individual diagnostic reactions with high-resolution melting (HRM) analysis.

Purpose of the Study:

To develop QMC-PCRx, a modified QMC-PCR protocol with a multiplexed second stage for simultaneous testing of multiple targets.
To assess the diagnostic accuracy and efficiency of QMC-PCRx compared to standard QMC-PCR.

Main Methods:

The PDM reaction was unchanged. The second stage was redesigned for multiplex HRM (mHRM) analysis using in silico design.
Seventeen colorectal cancer samples were tested for mutations in five hotspots using QMC-PCR (singleplex) and QMC-PCRx (multiplexed combinations).
Agreement between methods was determined by percentage concordance.

Main Results:

Multiplex HRM optimization allowed for distinct, non-overlapping peaks with up to three targets per reaction.
QMC-PCRx reduced the number of second-stage tests from 85 to 34 for 85 individual targets.
A 100% concordance was observed between the QMC-PCRx and standard QMC-PCR methodologies.

Conclusions:

Multiplexed analysis using HRM is feasible without compromising diagnostic accuracy.
The QMC-PCRx protocol offers a significant reduction in workload and cost for mutation screening.