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Real Time RT-PCR

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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.
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Updated: May 29, 2026

TChIP-Seq: Cell-Type-Specific Epigenome Profiling

TChIP-Seq: Cell-Type-Specific Epigenome Profiling

Published on: January 23, 2019

GETPrime: a gene- or transcript-specific primer database for quantitative real-time PCR.

Carine Gubelmann¹, Alexandre Gattiker, Andreas Massouras

¹Institute of Bio-engineering, School of Life Sciences, Laboratory of Systems Biology and Genetics, Lausanne, Switzerland.

Database : the Journal of Biological Databases and Curation

|September 16, 2011

Summary

This summary is machine-generated.

GETPrime is a novel database and platform that automates the design of highly specific primers for quantitative real-time polymerase chain reaction (qPCR). This tool enables accurate profiling of alternative splice variants, improving the study of gene expression.

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Area of Science:

Molecular Biology
Genomics
Bioinformatics

Background:

Alternative splicing generates numerous splice variants, but their functions remain poorly understood due to a lack of sensitive expression profiling tools.
Quantitative real-time polymerase chain reaction (qPCR) is a sensitive method for nucleic acid quantification, including splice variants.
Existing primer design tools lack specificity for splice variants and vary in quality and functionality.

Purpose of the Study:

To introduce GETPrime, a novel database and automated platform for designing optimal qPCR primers.
To enable gene-specific or transcript-specific expression profiling of splice variants.
To facilitate the experimental validation of primer specificity and expression patterns.

Main Methods:

Development of a novel platform integrating automated features for qPCR primer design.
Consideration of all gene splice variants for primer selection.
Primer specificity validation and automated selection of best primer pairs based on strict criteria.
Graphical visualization of primer pairs within their genomic context.

Main Results:

GETPrime successfully designs primers that distinguish between splice variants.
Experimental validation confirmed high transcript specificity of GETPrime primers in complex biological samples.
The platform automates primer design, specificity validation, and visualization.

Conclusions:

GETPrime provides a user-friendly, free-access database for efficient primer retrieval and visualization.
This tool significantly advances the ability to study splice variant expression profiles with high sensitivity.
GETPrime supports research on alternative splicing in common model organisms.