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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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qPCRTag Analysis - A High Throughput, Real Time PCR Assay for Sc2.0 Genotyping

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QPCR: Application for real-time PCR data management and analysis.

Stephan Pabinger¹, Gerhard G Thallinger, René Snajder

¹Institute for Genomics and Bioinformatics, Graz University of Technology, Petersgasse 14, 8010 Graz, Austria. stephan.pabinger@tugraz.at

BMC Bioinformatics

|August 29, 2009

Summary

A new web-based application called QPCR streamlines quantitative real-time polymerase chain reaction (qPCR) data analysis. This tool integrates storage, management, and multiple analysis methods for comprehensive gene expression quantification.

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Area of Science:

Molecular Biology
Bioinformatics

Background:

Quantitative real-time polymerase chain reaction (qPCR) is the standard for gene expression quantification.
Current qPCR data analysis involves multiple applications, lacking a unified solution.
Increasing sample sizes necessitate efficient and integrated analysis tools.

Purpose of the Study:

To develop a single, web-based application for the complete qPCR data analysis workflow.
To integrate data storage, management, and diverse analysis methods for qPCR experiments.

Main Methods:

Developed a Java-based, web-accessible application named QPCR.
Implemented data import functionality for qPCR instrument outputs.
Incorporated various analysis modules for cycle-threshold and efficiency calculations.

Main Results:

The QPCR application facilitates storage, management, and analysis of qPCR data.
It supports replicate handling, normalization, efficiency incorporation, and inter-run calibration.
Error propagation assessment and statistical testing on replicates are included, with customizable result visualization and export.

Conclusions:

A comprehensive web-based system, QPCR, has been developed to enhance qPCR experiment analysis.
The application integrates parsing, analysis, and chart generation into a single platform.
The QPCR system is freely available for use.