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

51.7K
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...
51.7K
Quantitative Analysis01:12

Quantitative Analysis

1.7K
Quantitative analysis is a technique for measuring the amount of specific constituents in a sample. When the sample's composition is unknown, qualitative analysis is performed first to identify its components, which ensures that the correct substances are measured during the quantitative phase.
In quantitative analysis, two key measurements are made: the sample quantity and a property proportional to the amount of the analyte (the substance being analyzed). This forms the basis of the...
1.7K

You might also read

Related Articles

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

Sort by
Same author

Full-Length 16S and 18S rRNA Long-Read Sequencing Reveals Gut Microbiome Diversity in the European Brown Hare (Lepus europaeus).

Environmental microbiology reports·2026
Same author

RNA-Based Biomarkers for Diagnostic Discrimination of Ischemic and Hemorrhagic Stroke: A Systematic Review.

Journal of clinical medicine·2026
Same author

Serum NFL and neuropsychological performance over ∼8 years in women with and without HIV: a longitudinal repeated measures study.

EClinicalMedicine·2025
Same author

Blood-based protein biomarkers during the acute ischemic stroke treatment window: a systematic review.

Frontiers in neurology·2024
Same author

Tachykinins and the potential causal factors for post-COVID-19 condition.

The Lancet. Microbe·2023
Same author

Oral hygiene, mouthwash usage and cardiovascular mortality during 18.8 years of follow-up.

British dental journal·2023

Related Experiment Video

Updated: Apr 26, 2026

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
10:50

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

16.0K

A method for quantitative analysis of standard and high-throughput qPCR expression data based on input sample

Mateusz G Adamski1, Patryk Gumann2, Alison E Baird3

  • 1Neurology, SUNY Downstate Medical Center, Brooklyn, New York, United States of America; Neurology, UJCM, Krakow, Poland.

Plos One
|August 5, 2014
PubMed
Summary

A new gene expression analysis method offers accurate quantification using input sample quantity, independent of control genes. This method is efficiency-corrected and normalized using a universal reference, proving valuable for clinical studies.

More Related Videos

Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass
14:29

Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass

Published on: May 1, 2013

13.8K
Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples
07:30

Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples

Published on: June 8, 2020

11.5K

Related Experiment Videos

Last Updated: Apr 26, 2026

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards
10:50

Single-cell Gene Expression Profiling Using FACS and qPCR with Internal Standards

Published on: February 25, 2017

16.0K
Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass
14:29

Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass

Published on: May 1, 2013

13.8K
Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples
07:30

Optimization for Sequencing and Analysis of Degraded FFPE-RNA Samples

Published on: June 8, 2020

11.5K

Area of Science:

  • Molecular Biology
  • Genomics

Background:

  • Quantitative polymerase chain reaction (qPCR) is the gold standard for gene expression analysis.
  • High-throughput qPCR enables simultaneous analysis of thousands of reactions, increasing sensitivity.
  • Existing methods often rely on control genes and can be sensitive to variations in amplification efficiency.

Purpose of the Study:

  • To present a novel gene expression analysis method adaptable for both standard and high-throughput qPCR.
  • To develop a method that is independent of control gene expression and accounts for input sample quantity.
  • To enable accurate and reproducible gene expression quantification across different batches and instruments.

Main Methods:

  • The input quantity method adjusts for the number of cells or input RNA.
  • The method incorporates efficiency correction and normalization using a universal reference complementary DNA (cDNA).
  • The method allows for absolute quantification and a non-efficiency corrected analysis as modifications.

Main Results:

  • The input quantity method demonstrated validity when compared to other common algorithms.
  • The method provides efficiency-corrected, normalized gene expression data.
  • Results are reproducible across different batches and instruments, irrespective of amplification efficiency variations.

Conclusions:

  • The presented gene expression analysis method is robust and accurate for both standard and high-throughput qPCR.
  • This technique is particularly valuable for clinical research involving whole blood and circulating leukocytes due to readily available cell counts.
  • The method enhances the reliability of gene expression studies by minimizing batch and instrument-specific variations.