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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...

RACE - Rapid Amplification of cDNA Ends

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...

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Related Experiment Video

Updated: Nov 11, 2025

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR

Published on: May 16, 2012

Gene Expression Analysis by Reverse Transcription Quantitative PCR.

Eva M Campion¹, Sinéad T Loughran²

¹Department of Life Science, Faculty of Science, Institute of Technology Sligo, Ash Lane, Co. Sligo, Ireland. campion.eva@itsligo.ie.

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

|March 25, 2021

Summary

This summary is machine-generated.

Reverse transcription quantitative polymerase chain reaction (RT-qPCR) offers rapid detection of RNA transcripts. This protocol details RT-qPCR for analyzing gene expression in Helicobacter pylori or infected samples.

Keywords:

RNA Real-time PCR Relative quantification Reverse transcription cDNA

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Last Updated: Nov 11, 2025

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Area of Science:

Microbiology
Molecular Biology
Infectious Diseases

Background:

Helicobacter pylori (H. pylori) is a significant human pathogen.
Gene expression analysis is crucial for understanding host-pathogen interactions.
Reverse transcription quantitative polymerase chain reaction (RT-qPCR) is a sensitive method for RNA quantification.

Purpose of the Study:

To present a two-step protocol for gene expression analysis.
To enable the study of H. pylori gene expression.
To facilitate the investigation of host responses to H. pylori infection.

Main Methods:

Utilizes reverse transcription quantitative polymerase chain reaction (RT-qPCR).
Employs a two-step protocol for RNA analysis.
Applicable to H. pylori and infected host samples.

Main Results:

The protocol allows for specific amplification and quantification of RNA transcripts.
RT-qPCR enables detection and quantification of H. pylori.
Host gene expression in response to infection can be monitored.

Conclusions:

RT-qPCR is a valuable tool for H. pylori research.
The outlined protocol provides a method for analyzing gene expression in relevant samples.
This technique aids in understanding H. pylori infection dynamics and host responses.