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Main focus of the GENE
QUANTIFICATION web page is, to describe and summarize all technical
The GENE QUANTIFICATION web page illustrates the usefulness of reliable quantification strategies, e.g. absolute-, relative-quantification assays, and the difference between them in kinetic PCR & kinetic RT-PCR.
RT-PCR is the technique of
choice for analyzing mRNA in extremely low abundance. But real-time
To obtain high fidelity,
accuracy and reliability in reverse transcription and subsequent kinetic
Main focus of the PHYSIOLOGY
web page is, to describe all kind of kinetic RT-PCR applications in
The reverse transcription - polymerase chain reaction (RT-PCR) is the most sensitive method for the detection of low abundance mRNA, often obtained from limited tissue samples. However, real-time RT-PCR is a complex technique, there are substantial problems associated with its true sensitivity, reproducibility and specificity and, as a quantitative method, it suffers from the problems inherent in RT and PCR. The recent introduction of fluorescence based kinetic (real-time) RT-PCR procedures significantly simplifies the process of producing reproducible quantification of mRNAs and promises to overcome these limitations. Nevertheless, their successful application depends on a clear understanding of the practical problems, and careful experimental design, application and validation remain essential for accurate quantitative measurements of the transcriptom.
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