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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...

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Related Experiment Video

Updated: Jul 4, 2026

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR

Published on: February 6, 2026

RNA isolation and real-time quantitative RT-PCR.

Haiyan Guan¹, Kaiping Yang

¹Children's Health Research Institute and Lawson Health Research Institute, Department of Obstetrics and Gynaecology, The University of Western Ontario, London, Ontario, Canada.

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

|June 3, 2008

Summary

Adipose tissue is an endocrine organ. This study presents specialized RNA isolation and real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) methods for analyzing messenger ribonucleic acid (mRNA) in adipose tissue.

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Area of Science:

Endocrinology
Molecular Biology
Gene Expression Analysis

Background:

Adipose tissue functions as a critical endocrine organ, secreting hormones and factors that influence systemic metabolism.
Gene activity regulation occurs at transcriptional and post-transcriptional levels, with messenger ribonucleic acid (mRNA) levels indicating gene expression.
Accurate mRNA quantification is essential for understanding gene activity within various tissues.

Purpose of the Study:

To present specialized techniques for isolating RNA from adipose tissue and adipocytes.
To describe a reliable and sensitive method for quantifying mRNA levels in adipose tissue.
To facilitate the study of gene expression in this important endocrine organ.

Main Methods:

Development and testing of unique RNA isolation protocols for lipid-rich adipose tissue and isolated adipocytes.
Application of real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) for mRNA analysis.
Validation of methods in rodent models (rats and mice).

Main Results:

Established effective RNA isolation procedures tailored to the challenges of adipose tissue.
Demonstrated the sensitivity and reliability of the RT-qPCR protocol for mRNA quantitation.
Provided a robust methodology for studying gene expression in adipose tissue.

Conclusions:

Specialized RNA isolation techniques are necessary for accurate analysis of adipose tissue.
Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) is a superior method for mRNA quantitation in this context.
The presented methods enable comprehensive investigation of adipose tissue's role in metabolic regulation.