Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

67.1K
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...
67.1K

You might also read

Related Articles

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

Sort by
Same author

Association of Markers of Kidney Tubular Secretion and Risk of Acute Kidney Injury after Acute Heart Failure.

Kidney360·2026
Same author

Markers of kidney tubule dysfunction and injury and long-term risk of acute kidney injury following coronary artery bypass graft surgery.

PloS one·2026
Same author

Electrocontrolled Injection-Coupled Droplet Microfluidic Platform for Antimicrobial Resistance Screening.

Analytical chemistry·2026
Same author

DELP-Net: A Differentiable Entropy Layer Pyramid Network for End-to-End Low-Rate DoS Detection.

Entropy (Basel, Switzerland)·2026
Same author

Corrigendum to "The deficiency of ALKBH5 promotes lenvatinib resistance and CD8<sup>+</sup> T cell exhaustion via accumulation of the cholesterol metabolite 27HC in hepatocellular carcinoma" [643 10 (2026) 218316].

Cancer letters·2026
Same author

Ionomer Structure Engineering for Optimal Triple-Phase Boundaries in Anion Exchange Membrane Water Electrolysers.

ACS applied materials & interfaces·2026

Related Experiment Video

Updated: Apr 14, 2026

Investigating Drivers of Antireward in Addiction Behavior with Anatomically Specific Single-Cell Gene Expression Methods
09:29

Investigating Drivers of Antireward in Addiction Behavior with Anatomically Specific Single-Cell Gene Expression Methods

Published on: August 4, 2022

2.9K

Simple regression for correcting ΔCt bias in RT-qPCR low-density array data normalization.

Xiangqin Cui1, Shaohua Yu2, Ashutosh Tamhane3

  • 1Department of Biostatistics, School of Public Health, University of Alabama at Birmingham, Birmingham, AL, 35294, USA. xcui@uab.edu.

BMC Genomics
|April 19, 2015
PubMed
Summary
This summary is machine-generated.

A new regression method corrects bias in reverse transcription quantitative PCR (RT-qPCR) data normalization. This approach improves the accuracy of gene expression analysis, particularly for rheumatoid arthritis studies.

More Related Videos

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays
10:58

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays

Published on: December 3, 2010

18.0K
Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR
14:14

Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR

Published on: December 6, 2014

17.5K

Related Experiment Videos

Last Updated: Apr 14, 2026

Investigating Drivers of Antireward in Addiction Behavior with Anatomically Specific Single-Cell Gene Expression Methods
09:29

Investigating Drivers of Antireward in Addiction Behavior with Anatomically Specific Single-Cell Gene Expression Methods

Published on: August 4, 2022

2.9K
Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays
10:58

Profiling of Pre-micro RNAs and microRNAs using Quantitative Real-time PCR qPCR Arrays

Published on: December 3, 2010

18.0K
Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR
14:14

Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR

Published on: December 6, 2014

17.5K

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Reverse transcription quantitative PCR (RT-qPCR) is a standard method for gene expression quantification.
  • Current normalization relies on the delta Ct (ΔCt) method, comparing target and reference genes.
  • This method can be susceptible to biases from varying PCR amplification efficiencies.

Purpose of the Study:

  • To identify and address bias in RT-qPCR data normalization.
  • To develop a more accurate method for analyzing gene expression data.

Main Methods:

  • Analysis of rheumatoid arthritis RT-qPCR low-density array datasets.
  • Examination of public mRNA and miRNA RT-qPCR array datasets.
  • Development of a per-gene regression method to adjust reference gene Ct values.

Main Results:

  • The standard ΔCt normalization method introduces significant bias due to differences in PCR amplification efficiency.
  • This bias leads to inaccurate fold change estimations and affects differential gene expression analysis.
  • The proposed regression method effectively removes the ΔCt bias.

Conclusions:

  • The per-gene regression method provides accurate normalization for RT-qPCR data.
  • This improved method can be applied to both low-density arrays and individual RT-qPCR assays.
  • It enhances the reliability of gene expression studies.