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Real-Time PCR: Current Technology and Applications

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ISBN-10:: 1904455395
ISBN-13:: 9781904455394
Pub. Date:: 01/01/2009
Publisher:: Caister Academic Press Limited

ISBN-10:: 1904455395
ISBN-13:: 9781904455394
Pub. Date:: 01/01/2009
Publisher:: Caister Academic Press Limited

Real-Time PCR: Current Technology and Applications

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$319.00

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Overview

Real-time PCR (RT-PCR) technology is highly flexible and many alternative instruments and fluorescent probe systems have been developed recently. The decreased hands-on time, increased reliability, and improved quantitative accuracy of RT-PCR methods have contributed to the adoption of RT-PCR for a wide range of new applications. This essential manual presents a comprehensive guide to the most up-to-date technologies and applications, as well as providing an overview of the theory of this increasingly important technique. Renowned experts in the field describe and discuss the latest PCR platforms, fluorescent chemistries, validation software, data analysis, and internal and external controls. This timely and authoritative volume also discusses a wide range of RT-PCR applications including clinical diagnostics, biodefense, RNA expression studies, validation of array data, mutation detection, food authenticity and legislation, NASBA, molecular halotyping, and much more. Real-Time PCR: Current Technology and Applications will be an essential book for all laboratories using PCR.

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Product Details

ISBN-13:	9781904455394
Publisher:	Caister Academic Press Limited
Publication date:	01/01/2009
Pages:	290
Product dimensions:	7.10(w) x 9.90(h) x 0.90(d)

List of contributors v

Preface ix

1 An Introduction to Real-Time PCR Nick A. Saunders 1

2 An Overview of PCR Platforms Julie M. J. Logan Kirstin J. Edwards 7

3 Homogeneous Fluorescent Chemistries for Real-Time PCR Martin A. Lee David J. Squirrell Dario L. Leslie Tom Brown 23

4 Reference Gene Validation Software for Improved Normalization Jo Vandesompele Mikael Kubista Michael W. Pfaffl 47

5 Data Analysis Software Michael W. Pfaffl Jo Vandesompele Mikael Kubista 65

6 Performing Real-time PCR Kirstin J. Edwards Julie M. J. Logan 85

7 Internal and External Controls for Reagent Validation Martin A. Lee Dario L. Leslie David J. Squirrell 95

8 Introduction to the Applications of Real-Time PCR Nick A. Saunders 105

9 Analysis of mRNA Expression by Real-time PCR Stephen A. Bustin Tania Nolan 111

10 Validation of Array Data Elisa Wurmbach 137

11 Mutation Detection by Real-Time PCR Elaine Lyon Rong Mao Jeffrey Swensen 149

12 Real-Time NASBA Julie D. Fox Catherine Moore Diana Westmoreland 163

13 Applications in Clinical Microbiology Andrew David Sails 177

14 Diagnosis of Invasive Fungal Infections David S. Perlin 221

15 Biodefence Christina Egan Nick M. Cirino Kimberlee A. Musser 235

16 Real-time PCR: Application to Food Authenticity and Legislation Gordon Wiseman 253

17 Molecular Haplotyping by Real-time PCR Genevieve Pont-Kingdon Alison Millson Elaine Lyon 269

Index 279

Plates A-1

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Editorial Reviews

Reviewer: Ruben Mestril, PhD (Loyola University Medical Center)
Description: This book covers the many emerging uses of real-time polymerase chain reaction (RT-PCR) in both diagnostic and basic research. In the last several years, novel chemistries and instrumentation platforms have enabled the detection of PCR products on a real-time basis and led to the adoption of real-time RT-PCR as the preferred method of measuring gene expression in a quantitative manner. In addition, real-time RT-PCR is now the method of choice to validate results obtained from array analysis. The book is a compilation of 17 chapters dealing with subjects of both the application and analysis of real-time PCR. The editors have recruited for this task a team of international experts from a number of different institutions. This volume should be of utmost interest to all investigators interested and involved in using RT-PCR.
Purpose: The main purpose is to present the most current applications for real-time PCR and some updated techniques. The book provides a comprehensive picture of its use in the basic research setting where real-time PCR is mostly used to provide quantitative measurements of gene transcription. The technology is currently used to determine the expression of a particular gene changes in response to the administration of a drug, cell differentiation, or changes in environmental conditions. Real-time PCR also is widely used to rapidly detect nucleic acids that serve as a diagnostic for the occurrence of cancer or genetic abnormalities. Furthermore, the use of real-time PCR assays in the clinical microbiology laboratory has significantly improved the diagnosis of infectious diseases. The book provides clear examples of these diverse uses of the technology.
Audience: Mainly written for practicing scientists interested or involved in the use of real-time PCR, the book provides a comprehensive selection of the most recently developed applications for it. It is clear that the RT-PCR protocols covered in this book will be of interest to most, if not all, investigators engaged in research that uses this important technique.
Features: Among the most interesting applications are covered in chapter 9, which is dedicated to the presentation of new protocols, enzymes, chemistries, and instrumentation for the use of this application. Chapter 12 concentrates on nucleic acid sequence-based amplification (NASBA), a method used for the detection and quantification of genomic, ribosomal, or mRNA. A useful set of applications of real-time PCR in the clinical setting, for biodefense purposes, and in the food industry are also presented. This results in a well balanced book covering the many potential uses of real-time PCR.
Assessment: This book offers a good balance of the different applications of this powerful technique. It also provides a number of updated protocols for investigators interested in incorporating this technique in their research. In all chapters, the layout of the protocol is straightforward and based on proven examples that render credibility to the diverse methods. The book should prove to be valuable for all those interested in RT-PCR.

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