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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...

PCR

PCR

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: May 9, 2026

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

Published on: April 8, 2017

Using PCR to Target Misconceptions about Gene Expression.

Leslie K Wright¹, Dina L Newman

¹Thomas H. Gosnell School of Life Sciences, Rochester Institute of Technology, Rochester, NY 14623.

Journal of Microbiology & Biology Education

|July 17, 2013

Summary

This summary is machine-generated.

This lab exercise uses PCR to teach biology students the difference between genes and gene expression, correcting misconceptions by showing that genes exist even when not expressed.

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Last Updated: May 9, 2026

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Area of Science:

Molecular Biology
Genetics Education

Background:

Many introductory biology students struggle to differentiate between genes (DNA) and gene expression (mRNA/protein).
A common misconception is that genes are only present when actively expressed.

Purpose of the Study:

To present a PCR-based laboratory exercise for first- and second-year biology students.
To address and correct common misconceptions regarding gene expression and the presence of genes.

Main Methods:

Students transform E. coli with an inducible GFP gene plasmid.
They observe induced and un-induced colonies.
A PCR experiment is conducted to challenge students' predictions about gene presence.

Main Results:

The laboratory exercise effectively creates cognitive dissonance, as PCR results contradict initial misconceptions.
Field testing demonstrated significant learning gains in both knowledge and application of gene and gene expression concepts.

Conclusions:

This PCR-based exercise is an effective tool for teaching the distinction between genes and gene expression.
The hands-on approach helps students overcome fundamental misunderstandings in molecular biology.