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Transcript quantification methodology has evolved from Northern blots to RNase protection to competitive RT‐PCR. Each of these methods made strides toward greater sensitivity and accuracy in quantification. Since the introduction of the 7700 Sequence Detection System and Taqman ® chemistry by Applied Biosystems in 1996, quantitative real‐time PCR has become widely accepted as the most sensitive and accurate method for the quantification of both RNA and DNA from a wide variety of sources. Today, there are a number of real‐time instruments from a variety of vendors and several detection methodologies and kit chemistries available. At the same time, prices for real‐time platforms are falling dramatically. This trend means that the technology is moving from one found primarily in core laboratories to the bench tops of individual investigators. There are several ways to interrogate a cell for changes induced by artificial or natural agents during a biological process.
One way is to look for changes in cellular transcript levels that may indicate downstream changes in the corresponding protein. In another instance, the focus may be on the presence or absence of a viral or bacterial pathogen. In this case, detecting not only the presence but also the degree of infection provides valuable information. Alternatively, looking for the presence or the level of expression from a transgene or the inhibition of expression of an endogenous gene by an RNAi agent may be the question of interest. In all cases, quantitative real‐time PCR technology can be utilized to provide the required results. However, successful implementation of the technology requires the user to have a basic background in the theoretical principles of real‐time PCR as well as their practical application.
The goal of this review is to provide this information. It will not be possible to cover all aspects of real‐time PCR in this review. For a more comprehensive overview of real‐time PCR, “A‐Z of Quantitative PCR,” edited by Stephen Bustin, is highly recommended.
Abstract: This book is a comprehensive manual to allow both the novice researcher and the expert to set up and carry out quantitative PCR assays from scratch. However, this book also sets out to explain as many features of qPCR as possible, provide alternative viewpoints, methods, and aims to simulate the researchers into generating, interpreting, and publishing data that are reproducible, reliable, and biologically meaningful Written by the best experts in drug design from different countries. Edited by Stephen A. Bustin, Barts and The London Queen Mary's School of Medicine and Dentistry, London, United Kingdom.
Reviews Gregory L. Shipley, Ph.D., Director, Quantitative Genomics Core Laboratory, Department of Integrative Biology and Pharmacology.
The University of Texas Health Science Center, Houston The book, A-Z of Quantitative PCR edited by Dr. Stephen Bustin is, for once, exactly what the title claims, Everything you want and need to know about real-time quantitative PCR. Since its' inception in 1996, real-time quantitative PCR has grown dramatically in the assortment of available hardware, chemistries and uses for this extremely robust technology. However there has been no single reference to point new users to that completely covered all facets of this technology.
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The A-Z of Quantitative PCR fills that void. The material is presented in a very readable form for those new to the technique and yet has plenty of detailed information for experienced veterans. The book is devided into three sections beginning with three excellent overviews of the technique followed by several chapters covering the basic principles behind every aspect of preparing samples to performing the RT and PCRs to how the various assay chemistries and available instrumentation work. Finally, there are quite a few chapters devoted to specific applications of the technology.
A real plus are the appendices covering many handy biochemical facts and glossary of terms for the newly initiated. All in all a very readable and informative book that should be a must read for enyone wanting to get into the field of real-time quantitative PCR.
Ginzinger, Ph.D., Laboratory Director, Genome Analysis Core Facility, UCSF Comprehensive Cancer Center Dr. Stephen Bustin has done a remarkable job on thr aptly titled A-Z of Quantitative PCR. It has remarkable depth to satisfy the needs of the most experienced Q-PCR scientist as well as the breadth to inspire the novice. Very clearly written and easy to use protocols and helpful tips ensure success when attempting your first real-time Q-PCR experiment.
An amazingly rich resource for everything you wanted to know about real-time quantitative PCR-from the historical background to chemistried, instruments, and data analysis. This book will quickly become an essential reference manual for every lab wanting to perform quantitative PCR. If after reading this book and following recomendations you are not able to get Q-PCR to work, you should seriously consider changing careers.