Analysis of mRNA Expression by Real-Time PCR

Authors: Stephen A. Bustin and Tania Nolan

Abstract: The last few years have seen the transformation of the fluorescence- based real-time reverse transcription polymerase chain reaction (RT-PCR) from an experimental tool into a mainstream scientific technology. Assays are simple to perform, capable of high throughput, and combine high sensitivity with exquisite specificity. The technology is evolving rapidly with the introduction of new enzymes, chemistries and instrumentation and has become the "Gold Standard" for a huge range of applications in basic research, molecular medicine, and biotechnology. Nevertheless, there are considerable pitfalls associated with this technique and successful quantification of mRNA levels depends on a clear understanding of the practical problems and careful experimental design, application and validation.

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See also: Current PCR books