PCR Guru: An Ultimate Benchtop Reference for Molecular Biologists
By Ayaz Najafov and Gerta Hoxhaj
4/5
()
About this ebook
PCR Guru: An Ultimate Benchtop Reference for Molecular Biologists is provides researchers in molecular biology with a handy reference for approaching and solving challenging problems associated with PCR setup and optimization. As a laboratory guide, it emphasizes the technical aspects of employing PCR as a tool in molecular biology laboratories. The book covers the history of PCR and the basic science underlying it. It then discusses PCR at the bench level, starting with detailed description and tips on primer design, and continuing with the standard protocols used to perform PCR.
- Provides troubleshooting tips for various types of modifications of standard protocols
- Contains unique “Good Practices and Tips that are indispensable for the beginner and expert alike
- Features “Special Cases with applications of PCR, optimization, and troubleshooting
- Includes detailed appendices with tables, figures, and key protocols
- Organized as a systematic, concentrated resource to save time when addressing a PCR problem
Ayaz Najafov
Ayaz was born in Baku, Azerbaijan. He obtained his BSc and MSc degrees in Molecular Biology and Genetics from Bosphorus University, Istanbul, Turkey and got a 2-year training in Biochemistry as an intern at Baylor College of Medicine, Houston, Texas, USA. Ayaz pursued his PhD in Biochemistry at MRC Protein Phosphorylation Unit at Dundee University, Scotland, UK in Prof. Dario Alessi’s lab. Ayaz is currently a postdoctoral fellow in Prof. Junying Yuan’s lab at Harvard Medical School, Cell Biology Department.
Related to PCR Guru
Related ebooks
Calculations for Molecular Biology and Biotechnology Rating: 3 out of 5 stars3/5Understanding PCR: A Practical Bench-Top Guide Rating: 5 out of 5 stars5/5Diagnostic Molecular Biology Rating: 0 out of 5 stars0 ratingsChallenges in Delivery of Therapeutic Genomics and Proteomics Rating: 0 out of 5 stars0 ratingsConcepts and Techniques in Genomics and Proteomics Rating: 0 out of 5 stars0 ratingsAdvances in Cell and Molecular Diagnostics Rating: 5 out of 5 stars5/5Metagenomics for Microbiology Rating: 5 out of 5 stars5/5Principles and Applications of Clinical Mass Spectrometry: Small Molecules, Peptides, and Pathogens Rating: 0 out of 5 stars0 ratingsIntroduction to Protein Mass Spectrometry Rating: 0 out of 5 stars0 ratingsRNA Therapeutics: The Evolving Landscape of RNA Therapeutics Rating: 0 out of 5 stars0 ratingsGenome Editing: A Practical Guide to Research and Clinical Applications Rating: 0 out of 5 stars0 ratingsSmall Molecule Drug Discovery: Methods, Molecules and Applications Rating: 0 out of 5 stars0 ratingsThe Design and Development of Novel Drugs and Vaccines: Principles and Protocols Rating: 0 out of 5 stars0 ratingsMass Spectrometry for the Clinical Laboratory Rating: 0 out of 5 stars0 ratingsMicrofluidics for Pharmaceutical Applications: From Nano/Micro Systems Fabrication to Controlled Drug Delivery Rating: 0 out of 5 stars0 ratingsMolecular Biology Techniques: An Intensive Laboratory Course Rating: 5 out of 5 stars5/5PCR Protocols: A Guide to Methods and Applications Rating: 2 out of 5 stars2/5Diagnostic Molecular Pathology: A Guide to Applied Molecular Testing Rating: 4 out of 5 stars4/5PCR Applications: Protocols for Functional Genomics Rating: 4 out of 5 stars4/5Molecular Biology Techniques: A Classroom Laboratory Manual Rating: 0 out of 5 stars0 ratingsPrinciples and Applications of Molecular Diagnostics Rating: 5 out of 5 stars5/5Clinical Genomics Rating: 5 out of 5 stars5/5Basic Methods in Molecular Biology Rating: 5 out of 5 stars5/5Biotechnology: A Laboratory Course Rating: 0 out of 5 stars0 ratingsCalculations for Molecular Biology and Biotechnology: A Guide to Mathematics in the Laboratory Rating: 0 out of 5 stars0 ratingsAdvanced Methods in Molecular Biology and Biotechnology: A Practical Lab Manual Rating: 5 out of 5 stars5/5Bioinformatics for Beginners: Genes, Genomes, Molecular Evolution, Databases and Analytical Tools Rating: 5 out of 5 stars5/5Genome Engineering via CRISPR-Cas9 System Rating: 5 out of 5 stars5/5Virology: A Laboratory Manual Rating: 5 out of 5 stars5/5
Biology For You
Fantastic Fungi: How Mushrooms Can Heal, Shift Consciousness, and Save the Planet Rating: 5 out of 5 stars5/5Anatomy and Physiology For Dummies Rating: 4 out of 5 stars4/5Dopamine Detox: Biohacking Your Way To Better Focus, Greater Happiness, and Peak Performance Rating: 3 out of 5 stars3/5Anatomy 101: From Muscles and Bones to Organs and Systems, Your Guide to How the Human Body Works Rating: 4 out of 5 stars4/5The Obesity Code: the bestselling guide to unlocking the secrets of weight loss Rating: 4 out of 5 stars4/5Sapiens: A Brief History of Humankind Rating: 4 out of 5 stars4/5Lifespan: Why We Age—and Why We Don't Have To Rating: 4 out of 5 stars4/5Ultralearning: Master Hard Skills, Outsmart the Competition, and Accelerate Your Career Rating: 4 out of 5 stars4/5Your Brain: A User's Guide: 100 Things You Never Knew Rating: 4 out of 5 stars4/5Homo Deus: A Brief History of Tomorrow Rating: 4 out of 5 stars4/5Peptide Protocols: Volume One Rating: 4 out of 5 stars4/5The Great Mortality: An Intimate History of the Black Death, the Most Devastating Plague of All Time Rating: 4 out of 5 stars4/5This Will Make You Smarter: 150 New Scientific Concepts to Improve Your Thinking Rating: 4 out of 5 stars4/5Emotional Blackmail: When the People in Your Life Use Fear, Obligation, and Guilt to Manipulate You Rating: 4 out of 5 stars4/5The Winner Effect: The Neuroscience of Success and Failure Rating: 5 out of 5 stars5/5Why We Sleep: Unlocking the Power of Sleep and Dreams Rating: 4 out of 5 stars4/5Gut: The Inside Story of Our Body's Most Underrated Organ (Revised Edition) Rating: 4 out of 5 stars4/5All That Remains: A Renowned Forensic Scientist on Death, Mortality, and Solving Crimes Rating: 4 out of 5 stars4/5Mother of God: An Extraordinary Journey into the Uncharted Tributaries of the Western Amazon Rating: 4 out of 5 stars4/5How Emotions Are Made: The Secret Life of the Brain Rating: 4 out of 5 stars4/5Other Minds: The Octopus, the Sea, and the Deep Origins of Consciousness Rating: 4 out of 5 stars4/5The Sixth Extinction: An Unnatural History Rating: 4 out of 5 stars4/5Genius Kitchen: Over 100 Easy and Delicious Recipes to Make Your Brain Sharp, Body Strong, and Taste Buds Happy Rating: 0 out of 5 stars0 ratingsJaws: The Story of a Hidden Epidemic Rating: 4 out of 5 stars4/5The Grieving Brain: The Surprising Science of How We Learn from Love and Loss Rating: 4 out of 5 stars4/5Woman: An Intimate Geography Rating: 4 out of 5 stars4/5The Blood of Emmett Till Rating: 4 out of 5 stars4/5The Woman Who Changed Her Brain: And Other Inspiring Stories of Pioneering Brain Transformation Rating: 4 out of 5 stars4/5A Crack In Creation: Gene Editing and the Unthinkable Power to Control Evolution Rating: 4 out of 5 stars4/5
Reviews for PCR Guru
3 ratings0 reviews
Book preview
PCR Guru - Ayaz Najafov
PCR Guru
An Ultimate Benchtop Reference for Molecular Biologists
Ayaz Najafov
Harvard Medical School, Department of Cell Biology, Boston, MA, United States
Gerta Hoxhaj
Harvard T.H. Chan School of Public Health, Department of Genetics and Complex Diseases, Boston, MA, United States
Table of Contents
Cover
Title page
Copyright
Dedications
Biography
Preface
Acknowledgments
Note to the Reader
Chapter 1: Introduction
Abstract
1.1. A bit of history
1.2. What is PCR?
1.3. PCR components
1.4. Exponential nature of PCR
1.5. Popularity of PCR
Chapter 2: Procedure
Abstract
2.1. Primer design
2.2. Standard PCR setup
2.3. Controls
2.4. Cycling parameters
Chapter 3: Good PCR Practices
Abstract
Chapter 4: Optimization and Troubleshooting
Abstract
4.1. Optimization rules
4.2. General optimization strategies
4.3. Troubleshooting specific problems
Chapter 5: Tips and Tricks
Abstract
Chapter 6: Special Cases
Abstract
Glossary and Abbreviations
Appendix A: Agarose Gel Electrophoresis Protocol
Appendix B: Silver Staining of DNA Gels
Appendix C: Plasmid Miniprep Protocol (by using QIAprep Spin Miniprep
Kit)
Appendix D: Restriction Enzyme Digestion Protocol
Appendix E: Protocol for DNA Concentration Estimation Using Hoechst 33258
Appendix F: Formulas
Appendix G: Common Primers for Sequencing, Promoters, Housekeeping Genes, and Reporters
Appendix H: Tags
Appendix I: Thermostable DNA polymerases
Appendix J: Example of a Daily PCR Log
Appendix K: Common Buffers and Solutions
Appendix L: Useful Numbers
Appendix M: Yeast Colony PCR
Useful Websites
References
Index
Copyright
Academic Press is an imprint of Elsevier
125 London Wall, London EC2Y 5AS, United Kingdom
525 B Street, Suite 1800, San Diego, CA 92101-4495, United States
50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States
The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom
Copyright © 2017 Elsevier Inc. All rights reserved.
No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions.
This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein).
Notices
Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary.
Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility.
To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein.
Library of Congress Cataloging-in-Publication Data
A catalog record for this book is available from the Library of Congress
British Library Cataloguing-in-Publication Data
A catalogue record for this book is available from the British Library
ISBN: 978-0-12-804231-1
For information on all Academic Press publications visit our website at https://www.elsevier.com/
Publisher: Sara Tenney
Acquisitions Editor: Sara Tenney
Editorial Project Manager: Fenton Coulthurst
Production Project Manager: Chris Wortley
Designer: Christian Bilbow
Typeset by Thomson Digital
Dedications
I dedicate this book to my father, Azad Najafov, who introduced me to the world of molecular biology and to my friend and mentor, İbrahim Barış, who taught me how to do my first PCR.
Ayaz Najafov
I dedicate this book to my family to whom I am eternally grateful for their constant support and care for me and my academic career.
Gerta Hoxhaj
Biography
Ayaz Najafov
Ayaz was born in Baku, Azerbaijan and as a high school student, participated in local and international biology Olympiads, where he had won several gold, silver, and bronze medals. He obtained his BSc and MSc degrees in Molecular Biology and Genetics from Bosphorus University, Istanbul, Turkey, and got a 2-year training in Biochemistry as an intern at Baylor College of Medicine, Houston, TX, United States. Ayaz pursued his PhD in Biochemistry at MRC Protein Phosphorylation Unit at Dundee University, Scotland, United Kingdom in Prof. Dario Alessi’s lab. Ayaz is currently a postdoctoral fellow in Prof. Junying Yuan’s lab at Harvard Medical School, Cell Biology Department.
Gerta Hoxhaj
Gerta was born in Fier, Albania. She obtained her BSc degree in Molecular Biology and Genetics, as well as a second major in Chemistry from Bosphorus University, Istanbul, Turkey. Gerta pursued her PhD in Biochemistry at MRC Protein Phosphorylation Unit at Dundee University, Scotland, United Kingdom in Prof. Carol MacKintosh’s lab. Gerta is currently a postdoctoral fellow in Prof. Brendan Mannings’s lab at Harvard T.H. Chan School of Public Health, Department of Genetics and Complex Diseases.
Preface
This book was written with an aim of providing researchers practicing molecular biology, whether at their beginner or expert stage, with a handy tool and reference for setting up, performing, optimizing, and troubleshooting polymerase chain reaction (PCR). We believe that this book will provide tremendous assistance for solving problems associated with challenging cases of PCR.
This book does not provide the reader with vast details of the PCR theory and the multiple applications of the technique. As this book is a laboratory guide, the emphasis here was made on the technical aspects of employing PCR as a tool in molecular biology laboratories and the issues were described in bench terms,
which makes the book more comprehendible to the end users of the technique.
General optimization and troubleshooting strategies, as well as a detailed