In view of the ubiquity of the use of standard RT-PCR, the main objective of this volume is to provide novel, useful applications of RT-PCR. These include some useful adaptions and application that are relevant to the wider research communities who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptions are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imbalances in specific mRNA expression between normal and diseased tissues. RT-PCR has afforded many opportunities in diagnostics, allowing sensitive detection of RNA viruses such as HIV and HCV. RT-PCR facilities many diverse techniques in research, including in situ localization of mRNA, antibody engineering, and cDNA cloning. In particular, this volume highlights how RT-PCR, microarray technology, HPLC and time-resolved fluorimetry. This volume will stimulate the reader towards diverse new ways in which this remarkable technique can facilitate the molecular aspects of their biomedical research.
Once a tedious, highly skilled operation, reverse-transcription polymerase chain reaction (RT-PCR) has become a routine and invaluable technique used in most laboratories. In RT-PCR Protocols, Second Edition, expert researchers fully update the technologies presented in the popular previous edition, such as competitive RT-PCR, nested RT-PCR, RT-PCR from single cells, and RT-PCR for cloning. In addition, newer technologies are also explored, including multiplex RT-PCR, RT-LATE-PCR, and the greatly advanced field of real-time quantitative RT-PCR, while recent advances in creating the optimum RT-PCR reaction, e.g. RNA extraction, primer design, and reverse transcription, end the book with their indispensable input. Written in the highly successful Methods in Molecular Biology™ series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.
User friendly and up-to-date, RT-PCR Protocols, Second Edition acts as a handy companion to scientists from numerous diverse backgrounds who wish to explore further the marvels of gene expression.