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The arrival of termed reverse transcription-PCR (RT-PCR) meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. The main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. Quantitative RT-PCR provides the ability to accurately measure changes/imbalances in specific mRNA expression between normal and diseased tissues. Because of its remarkable sensitivity, RT-PCR enables the detection of low-abundance mRNAs even at the level of individual cells. RT-PCR has afforded many opportunities in diagnostics, allowing sensitive detection of RNA viruses such as HIV and HCV. RT-PCR facilitates many diverse techniques in research, including in situ localization of mRNA, antibody engineering, and cDNA cloning. In particular, the present work highlights how RT-PCR complements other technological advances, such as laser-capture microdissection (LCM), real-time PCR, microarray technology, HPLC, and time-resolved fluorimetry. RT-PCR has become one of the most widely applied techniques in biomedical research, and has been a major boon to the molecular investigation of disease pathogenesis. Determination of the pathogenesis of diseases at the molecular level is already beginning to inform the design of new therapeutic strategies.