Ebook Quantitative real-time PCR - Methods and protocols: Part 1

Part 1 book "Quantitative real-time PCR - Methods and protocols" includes content: Minimum information necessary for quantitative real; time PCR experiments; selection of reliable reference genes for RT-QPCR analysis; introduction to digital PCR; mediator probe PCR - detection of real time PCR by label; free probes and a universal fluorogenic reporter,... and other contents.
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Ebook Quantitative real-time PCR - Methods and protocols: Part 1 Methods in Molecular Biology 1160 Roberto Biassoni Alessandro Raso Editors Quantitative Real-Time PCR Methods and Protocols METHODS IN M O L E C U L A R B I O LO G Y Series Editor John M. Walker School of Life Sciences University of Hertfordshire Hatfield, Hertfordshire, AL10 9AB, UK For further volumes: http://www.springer.com/series/7651 Quantitative Real-Time PCR Methods and Protocols Edited by Roberto Biassoni Laboratorio Medicina Molecolare, Dipartimento Medicina Traslazionale, Istituto Giannina Gaslini, Genova, Italy Alessandro Raso Laboratorio U.O.C Neurochirurgia, Istituto Giannina Gaslini, Genova, Italy Editors Roberto Biassoni Alessandro Raso Laboratorio Medicina Molecolare Laboratorio U.O.C Neurochirurgia Dipartimento Medicina Traslazionale Istituto Giannina Gaslini Istituto Giannina Gaslini Genova, Italy Genova, Italy ISSN 1064-3745 ISSN 1940-6029 (electronic) ISBN 978-1-4939-0732-8 ISBN 978-1-4939-0733-5 (eBook) DOI 10.1007 /978-1-4939-0733-5 Springer New York Heidelberg Dordrecht London Library of Congress Control Number: 2014936291 © Springer Science+Business Media New York 2014 This work is subject to copyright. All rights are reserved by the Publisher, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilms or in any other physical way, and transmission or information storage and retrieval, electronic adaptation, computer software, or by similar or dissimilar methodology now known or hereafter developed. Exempted from this legal reservation are brief excerpts in connection with reviews or scholarly analysis or material supplied specifically for the purpose of being entered and executed on a computer system, for exclusive use by the purchaser of the work. Duplication of this publication or parts thereof is permitted only under the provisions of the Copyright Law of the Publisher’s location, in its current version, and permission for use must always be obtained from Springer. Permissions for use may be obtained through RightsLink at the Copyright Clearance Center. Violations are liable to prosecution under the respective Copyright Law. The use of general descriptive names, registered names, trademarks, service marks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. While the advice and information in this book are believed to be true and accurate at the date of publication, 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. Printed on acid-free paper Humana Press is a brand of Springer Springer is part of Springer Science+Business Media (www.springer.com) Preface From the first report describing real-time PCR detection in 1993, the number of different applications has grown exponentially. Since quantitative PCR is the “gold standard” tech- nology to quantify nucleic acids, thousands of articles and books have been written on both its description and its practical use. Nowadays, it is a very accessible technique, but some pitfalls should be overcome in order to achieve robust and reliable analysis. In this book, our aim is to focus on the different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan. Such issues have been described in the first six chapters, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for low-level detection and to develop noninvasive diagnosis. Our hope is that a professional, endowed with the knowledge of some of the method- ological issues and of some of the applications, could devise new qPCR-based approaches related to his or her area of investigation. We have tried to cover the possible qPCR meth- ods, but of course we could not cover here all of the feasible applications. We are grateful to all of the colleagues who have contributed to the book with these manuscripts sharing their methods with the qPCR community. Genova, Italy Roberto Biassoni Alessandro Raso v Contents Preface. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . v Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix 1 Twenty Years of qPCR: A Mature Technology? . . . . . . . . . . . . . . . . . . . . . . . . ...