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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: Aug 7, 2025

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

Disclosing quantitative RT-PCR raw data during manuscript submission: a call for action.

Andreas Untergasser^1,2, Jan Hellemans³, Michael W Pfaffl⁴

¹Zentrum für Molekulare Biologie der Universität Heidelberg, Germany.

Molecular Oncology

|March 14, 2023

Summary

This summary is machine-generated.

Ensuring scientific data accuracy is crucial for reproducibility in RNA research. We recommend detailed methods, raw RT-qPCR data submission, and the new Real-time PCR Data Essential Spreadsheet Format (RDES) for transparency.

Keywords:

RNA RT-qPCR accuracy quantification

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Last Updated: Aug 7, 2025

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

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

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Area of Science:

Molecular Biology
Bioinformatics
Genomics

Background:

Growing concern over scientific data reproducibility across research fields.
Increased publications in RNA profiling and sequencing studies highlight the need for accurate quantification of coding and noncoding RNAs.

Purpose of the Study:

To address the challenges in RNA quantification reproducibility.
To propose standardized recommendations for improving data transparency and accuracy in RT-qPCR studies.

Main Methods:

Recommending accurate documentation of experimental procedures within the main text of publications.
Advocating for the submission of raw RT-qPCR data for all reported experiments.
Introducing the Real-time PCR Data Essential Spreadsheet Format (RDES) for unified data submission.

Main Results:

The proposed recommendations aim to enhance the reliability and transparency of RNA quantification data.
The RDES format provides a simple, unified structure for RT-qPCR raw data submission.

Conclusions:

Implementing these recommendations will improve the evaluation of data reproducibility in RNA research.
Standardized data reporting through RDES facilitates better scientific scrutiny and data sharing.