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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

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Related Experiment Video

Updated: Jun 14, 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

Primer design for RT-PCR.

Kelvin Li¹, Anushka Brownley

¹J. Craig Venter Institute, Rockville, MD, USA. kli@jcvi.org

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

|March 20, 2010

Summary

This summary is machine-generated.

Computational methods enhance primer design for Polymerase Chain Reaction (PCR) experiments. Analyzing potential issues like primer dimers and off-target amplification computationally improves experimental success and data accuracy.

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Last Updated: Jun 14, 2026

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

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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

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Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)

Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)

Published on: August 6, 2011

Area of Science:

Molecular Biology
Bioinformatics

Background:

Primer design is critical for Polymerase Chain Reaction (PCR) success.
Suboptimal primers lead to inefficient amplification and non-specific products.

Purpose of the Study:

To highlight the importance of computational analysis in primer selection.
To demonstrate how bioinformatics tools can predict and prevent PCR failure.

Main Methods:

Utilizing computational sequence analysis tools.
A priori detection of potential primer pair limitations.

Main Results:

Identification of factors like primer dimers, alternative products, and sequence variations.
Minimization of PCR failures through predictive analysis.

Conclusions:

Computational primer design significantly reduces experimental failure rates.
Bioinformatics tools ensure specificity and accuracy in PCR amplification, preventing misleading results.