Biotechnology A Laboratory Skills Course – Brown J Kirk

Invention of PCR ? What Is PCR? ? Thermal Cyclers ? Types of PCR ? PCR Optimization ? Techniques Based on PCR ? Real-World Applications of PCR ? Laboratory Activities Activity 6.1 STR PCR Analysis Activity 6.2 GMO Detection by PCR Activity 6.3 Detection of the Human PV92 Alu Insertion Summary The polymerase chain reaction (PCR) has revolutionized the study of living things.

Invented by Kary Mullis in 1983, PCR has been a springboard for molecular biology research. It is the basis of the Human Genome Project, modern forensic analysis, and genetic engineering. Using PCR, a small DNA sequence consisting of just a few hundred base pairs can be found within a genome of billions of base pairs. Billions of copies of the sequence can be generated, making the DNA sequences available for study and manipulation. Agriculture has been transformed by PCR with the advent of genetically modified crops.

Even more recently, cows and goats have been genetically engineered to produce pharmaceutical drugs in their milk, creating a new industry called pharming. PCR has made forensic analysis cheap, fast, and extremely accurate. Today’s DNA profiles have less than a one in a trillion chance of matching another random individual, providing law enforcement with a powerful tool to fight crime.

PCR has also been used to compare Neanderthal and human DNA to provide insights into how these populations interacted tens of thousands of years ago. The activities in this chapter use PCR to investigate DNA profiling, to detect genetic modifications in food, and to study human ancestry.

Invention of PCR It was on Highway 128 in California at mile marker 46.58 in April 1983 that Kary Mullis (see Figure 6.1) had an epiphany. He pulled off the road and sketched out the process that would later be known as the polymerase chain reaction (PCR). He envisioned the use of small pieces of DNA to bracket and replicate a section of DNA. Mullis was a chemist working at Cetus, one of the first biotech companies in the U.S.

(Cetus was acquired by Chiron Corporation in 1991, and Chiron was acquired by Novartis International AG in 2006.) Mullis ran a laboratory that made oligonucleotides (short, single strands of DNA) and was interested in methods for sequencing DNA.

Developmental Editors: Nebila Idris and Yolanda Kowalewski Assistant Editors: Erika Fong, Kristi DeCourcy, and Joann Lau Editorial Assistant: Melody Binay Art Director: Ken Shiokari Design and Illustrations: KSD-CA Printer: Walsworth Print Group Copyright © 2011 Bio-Rad Laboratories, Inc. 1000 Alfred Nobel Drive, Hercules, CA 94547. All rights reserved. Manufactured in the United States of America. This publication is protected by copyright and permission should be obtained from the publisher prior to any prohibited reproduction, storage in a retrieval system, or transmission in any form or by any means, electronic, mechanical, photocopying, recording, or likewise.

To obtain permission(s) to use material from this work, please submit a written request to Bio-Rad Laboratories, Biotechnology Explorer Program, 1000 Alfred Nobel Dr., Hercules, CA 94547. For information regarding permissions, please email [email protected]. ISBN 978-0-9832396-0-4 Printing number 10 9 8 7 6 5 4 3 2 Disclaimer Students, educators, and researchers using experimental procedures outlined in this book do so at their own risk. Users of this book are solely responsible for obtaining the necessary licenses, certificates, and permissions to conduct any experimental procedures that are subject to national or local legislation or restrictions.

Users of this book are responsible for the humane treatment of animals according to local and national guidelines. The author and Bio-Rad Laboratories, Inc. do not assume responsibility for failure of a user to do so. Ute Science Group 7661025 C ii Bio-Rad Laboratories, Inc. Web site www.bio-rad.com USA 800 424 6723 Austral/a 61 2 9914 2800 Austria 01 877 89 01 Belgium 09 385 55 11 Braz/I 55 11 5044 5699 Canada 905 364 3435 China 86 21 6169 8500 Czech Republic 420 241430 532 Denmark 44 52 10 00 Rn/and 09 804 22 00 France 01 47 95 69 65 Germany 089 31 884 O Greece 30 210 9532 220 Hong Kong 852 2789 3300 Hungary 36 1 459 6100 lnd/a91 124 4029300 Israel 03 963 6050 ltaly39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460 Mexlco 52 555 488 7670 The Netherlands 0318 540666 New Zealand 64 9 415 2280 Norway 23 38 41 30 Poland 48 22 331 99 99 Portugal 351 21 472 7700 Russia 7 495 721 14 04 Singapore 65 6415 3188 South Africa 27 861 246 723 Spain 34 91 590 5200 S-n 08 555 12700 Switzerland 061 717 95 55 Taiwan 886 2 2578 7189 Thailand 800 88 22 88 United Klngdom 020 8328 2000 11-0179 0312 Sig 1211 Dedication This book represents a lifetime of teaching experience and sacrifice.

I would like to dedicate it to a number of people. To my immediate family: my father and mother, Jim and Janice Brown, and my sister Lisa and her family, who fostered my natural curiosity into a lifetime of wonder about the world around me.

This is a short excerpt from the opening of “” by Unknown, quoted for review and introduction purposes. All rights belong to the copyright holders.

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