Rt-pcr Protocols

A panel of highly regarded molecular biologists and clinical researchers describe in detail their most novel, useful, and interesting RT-PCR applications. Here the newcomer will find readily reproducible protocols for highly sensitive detection and quantification of gene expression, the in situ localization of gene expression in tissue, and the cloning of genes, as well as for analyzing T-cell clones and the differential expression of genes. For the expert seeking to extend the usefulness of RT-PCR, there are user-friendly applications that complement the latest technological advances, including laser-capture microdissection (LCM), real-time and quantitative PCR, microarray technology, cDNA cloning, and antibody engineering.

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.