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PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

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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: May 19, 2026

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Quantitative PCR.

L Raeymaekers¹

¹Laboratorium Voor Fysiologie Ku Leuven, Campus Gasthuisberg, Leuven, Belgium.

Methods in Molecular Medicine

|March 11, 2011

Summary

This summary is machine-generated.

Quantitative PCR (polymerase chain reaction) is vital in clinical settings for detecting diseases and genetic changes. Accurately quantifying initial DNA amounts is challenging due to PCR

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Area of Science:

Molecular Biology
Clinical Diagnostics
Biotechnology

Background:

Quantitative PCR (polymerase chain reaction) is increasingly used in clinical investigations.
Applications include residual disease assessment, viral nucleic acid detection, and gene amplification/deletion analysis.
Reliable quantification of initial target template (T(o)) from PCR product (T(n)) remains a challenge.

Purpose of the Study:

To address the challenges in quantitative PCR (qPCR).
To improve the reliability of determining initial template amounts in qPCR assays.
To overcome limitations posed by PCR amplification and saturation.

Main Methods:

The study focuses on the principles and challenges of quantitative PCR (qPCR).
It discusses the relationship between initial template amount (T(o)) and PCR product accumulation (T(n)).
Key PCR properties influencing quantification, such as amplification factor and saturation, are examined.

Main Results:

The accuracy of qPCR is highly dependent on experimental conditions.
High amplification factors make PCR sensitive to minor variations.
Saturation phenomenon leads to decreased amplification efficiency in later cycles.

Conclusions:

Accurate quantitative PCR (qPCR) requires careful control of experimental conditions.
Understanding and mitigating the effects of amplification and saturation are crucial for reliable results.
Further research is needed to optimize qPCR protocols for clinical applications.