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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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Updated: May 29, 2026

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs

Published on: April 14, 2015

Quantitative rt-PCR.

¹Clontech Laboratories, Palo Alto, CA.

Methods in Molecular Medicine

|March 11, 2011

Summary

This summary is machine-generated.

Quantitative analysis using reverse transcriptase polymerase chain reaction (RT-PCR) is challenging due to its two enzymatic steps and PCR

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Last Updated: May 29, 2026

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Area of Science:

Molecular Biology
Biochemistry

Background:

Reverse transcriptase polymerase chain reaction (RT-PCR) is a sensitive method for mRNA analysis.
Quantitative analysis with RT-PCR presents significant challenges.

Purpose of the Study:

To identify the primary obstacles in obtaining quantitative data from RT-PCR.
To explore adaptations for achieving accurate quantitative results with RT-PCR.

Main Methods:

Analysis of the two sequential enzymatic steps in RT-PCR: DNA synthesis and PCR amplification.
Evaluation of practical challenges and the impact of PCR's exponential nature on quantification.

Main Results:

The dual enzymatic steps and the inherent exponential amplification of PCR complicate quantitative measurements.
Practical considerations in performing RT-PCR also hinder accurate quantification.

Conclusions:

Despite inherent difficulties, RT-PCR can be adapted for accurate quantitative mRNA analysis.
Modifications to the RT-PCR protocol are key to overcoming quantification challenges.