PCR Protocols
Known for flexibility and robustness, PCR techniques continue to improve through numerous developments, including the identification of thermostable DNA polymerases which exhibit a range of properties to suit given applications. PCR Protocols, Third Editi
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Molecular Biology™
Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK
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PCR Protocols Third Edition Edited by
Daniel J. Park Genetic Epidemiology Laboratory, Department of Pathology, The University of Melbourne, Parkville, VIC, Australia
Editor Daniel J. Park, Ph.D. (CANTAB) Genetic Epidemiology Laboratory Department of Pathology The University of Melbourne Parkville, VIC Australia [email protected]
ISSN 1064-3745 e-ISSN 1940-6029 ISBN 978-1-60761-943-7 e-ISBN 978-1-60761-944-4 DOI 10.1007/978-1-60761-944-4 Springer New York Dordrecht Heidelberg London Library of Congress Control Number: PCN applied for © Springer Science+Business Media, LLC 2011 All rights reserved. This work may not be translated or copied in whole or in part without the written permission of the publisher (Humana Press, c/o Springer Science+Business Media, LLC, 233 Spring Street, New York, NY 10013, USA), except for brief excerpts in connection with reviews or scholarly analysis. Use in connection with any form of information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed is forbidden. The use in this publication of trade names, trademarks, service marks, and similar terms, even if they are not identified as such, is not to be taken as an expression of opinion as to whether or not they are subject to proprietary rights. While the advice and information in this book are believed to be true and accurate at the date of going to press, neither the authors nor the editors nor the publisher can accept any legal responsibility for any errors or omissions that may be made. The publisher makes no warranty, express or implied, with respect to the material contained herein. Cover illustration: Schematic structure of Phusion® High-Fidelity DNA Polymerase, kindly provided by Finnzymes Oy, a Thermo Fisher Scientific Company. Printed on acid-free paper Humana Press is part of Springer Science+Business Media (www.springer.com)
Preface Since the concept of PCR was first described, PCR has taken on a central role in many areas of biological research and diagnostics. This continues today, including its application to the latest generation genome-wide analysis and high-throughput sequencing platforms. Although other methods have been described to enable the exponential amplification of nucleic acids, such as NASBA, LAMP, and LCR, PCR remains unparalleled in terms of its flexibility and robustness. From the basic concept of using a pair of primers and polymerase to amplify a defined region of nucleic acid, a multitude of derivative techniques and applications have emerged. Numerous developments have occurred to improve on early iterations of PCR, including the identification of thermostable DNA polymerases which exhibit a range of properties to suit given applications. The use of such enzymes offer tailore
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