Gene ticsLaboratory investigations

Fourteenth edition

t hom as R. meRtensBall State University

Robe Rt L. h a mmeRsmithBall State University

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Senior Acquisitions Editor: Michael GillespieProject Manager: Dorothy CoxAssistant Editor: Chloé VeylitProgram Manager: Anna AmatoProgram Management Team Lead: Mike EarlyProject Management Team Lead: Nancy TaborProduction Management: Integra Software Services Pvt. Ltd.Design Manager: Marilyn PerryCover Designer: Marilyn PerryInterior Designer: Integra Software Services Pvt. Ltd.Rights & Permissions Project Manager: Maya GomezRights & Permissions Management: Lumina DatamaticsManufacturing Buyer: Stacey WeinbergerExecutive Marketing Manager: Neena BaliCover Photo Credit: Robert L. HammersmithChapter Opener Photos Credit: Kenneth Eward/BioGrafx

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Library of Congress Cataloging-in-Publication Data

Mertens, Thomas Robert, 1930– Genetics : laboratory investigations/Thomas Mertens, Robert Hammersmith.—14th edition. pages cm ISBN 978-0-321-81417-3 1. Genetics—Laboratory manuals. I. Hammersmith, Robert L. II. Title. QH440.5.M47 2015 576.5078—dc23 2014021188

www.pearsonhighered.comISBN 10: 0-321-81417-7ISBN 13: 978-0-321-81417-3

1 2 3 4 5 6 7 8 9 10—V3N4—18 17 16 15 14

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iii

Preface v

1 Drosophila and maize experiments in Genetics: monohybrid and Dihybrid c rosses 1

2 Principles of Probability 25

3 t he c hi-square test 33

4 c ell Reproduction: mitosis 47

5 meiosis in a nimals: o ogenesis and spermatogenesis 55

6 meiosis in a ngiosperms: microsporogenesis 65

7 Polytene c hromosomes from Drosophila salivary Glands 79

8 sex c hromosomes and Gene t ransmission 85

9 t he sex c heck: a study of sex c hromatin in human c ells 95

10 human c hromosomes 101

11 Linkage and c rossing o ver 131

12 Genetics of a scospore c olor in Sordaria: a n investigation of Linkage and c rossing o ver Using tetrad a nalysis 151

13 o pen-ended experiments Using Drosophila: Locating a mutant Gene in its c hromosome 159

14 t he Genetic material: isolation of Dna 167

15 Restriction endonuclease Digestion and Gel electrophoresis of Dna 175

Content s

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iv c ontents

16 a mplification of Dna Polymorphisms by Polymerase c hain Reaction (Pc R) and Dna Fingerprinting 191

17 t ransformation of Escherichia coli 207

18 Gene a ction: synthesis of β-Galactosidase in Escherichia coli 217

19 c hromatographic c haracterization of Drosophila melanogaster mutants 223

20 bacterial mutagenesis 231

21 Gene Recombination in Phage 237

22 Polygenic inheritance: Fingerprint Ridge c ount 243

23 Population Genetics: t he hardy-Weinberg Principle 253

24 Population Genetics: t he effects of selection and Genetic Drift 265

25 a pplied human Genetics 275

26 ncbi and Genomic Data mining 295

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Pref a Ce

With the fourteenth edition, Genetics Laboratory Investigations is well into its seventh decade serving the needs of genetics instructors and students. Beginning in 1952, the first four edi-

tions were authored by Eldon Gardner of Utah State University. Thomas Mertens assumed the major responsibility for preparing the next three editions. Robert Hammersmith joined the writing team for the eighth edition; following Dr. Gardner’s death, Mertens and Hammersmith prepared editions nine through thirteen, and now the fourteenth. This long history of service to the genetics education com-munity was recognized in 1998 by the Text and Academic Authors Association with the presentation of the McGuffey Award.

Much of the success of the book can be accounted for by the user-friendly design meant to meet the needs of both faculty and students, and having been developed by those actively involved in the teaching of introductory general genetics at the undergraduate level. We are firmly convinced that a student’s laboratory experience in genetics is an essential part of the learning process and vitally important to fostering student interest in, and understanding of, the discipline. Toward this end, a number of “open-ended” investigations have been included in this manual—the most obvious being Investigation 13, “Open-Ended Experiments Using Drosophila: Locating a Mutant Gene in its Chromo-some.” With appropriate adaptation, however, instructors can use the open-ended approach in any of the investigations in which students collect original data.

n ew to this edition

We have tried to live up to the legacy of the manual in this new edition. As in previous editions, we have included investigations to accommodate courses taught in a variety of academic settings and to be cost-effective, especially for investigations involving molecular genetics. Many investigations provide two or more alternative ways of achieving the instructional objectives. Each investigation has been re-viewed and updated for newer data and clarity, as have the references at the end of each investigation. The fourteenth edition of the manual represents one of the most extensive revisions undertaken by the authors, and involves the conversion of most of the black and white photographs and diagrams to full color photographic plates and illustrations. This conversion has allowed us to now include many new photographs detailing differences in mutants of Drosophila, maize, barley and other organisms that previously could not have been adequately done with black and white illustrations.

• Over 85 new color photographs have been incorporated into 55 new or replacement figures, in-cluding new photographs of Drosophila and maize in Investigations 1, 8, 11, 13, and 19.

• Investigation 1 has been extensively rewritten to expand ideas for mono- and dihybrid crosses, with photographs of body, wing, bristle and eye-color mutants of Drosophila and various aleurone and starch characteristics in maize. The section on epistasis has also been expanded and detail photographs of maize aleurone epistasis provided, so that students can count kernels in the photo-graphs if actual epistatic ears are not available.

• New problem sets and problem examples for Investigations 2, 3, 22, and 23 have been added, with some problem sets drawing on data collected from maize ears featured in photographs in Investigation 1.

• Investigations 4, 5, and 6 have new color photographic plates of chromosomes in mitosis and meio-sis with a totally new plate on whitefish mitosis and ascaris meiosis.

v

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vi Preface

• New human chromosome spreads for student karyotyping have been add to Investigation 10.

• New photographs of tight linkage in maize and linkage in barley are found in Investigation 11, augmenting discussion of recombination and analysis of a problem set on the interpretation of F2 hybrid linkage data. Color figures of FISH (florescence in situ hybridization) have also been place in Investigation 11.

• Investigations 12, 14, 17, 20, 21, 22, and 24 have new color figures as well.

• We have undertaken a major revision of Investigation 26, “NCBI and Genomic Data Mining,” to ac-commodate changes in these federal data bases and it includes four new figures.

• A new Instructor’s Manual provides answers to most questions in each investigation, helpful hints for setting up experiments in certain investigations, and when needed, sources of instructional ma-terials beyond those suggested in the book itself.

To all who have assisted us over the years, we give our thanks. Credit for illustrations and tables in the present edition are given with appropriate investigations according to the wishes of the authors or publishers from whom permissions have been granted. Investigations 12 (Genetics of Sordaria), 22 (Human Fingerprint Ridge Counts), and 24 (Genetic Drift) were originally published in The Ameri-can Biology Teacher as was the portion of Investigation 6 dealing with meiosis in Tradescantia. We acknowledge with special appreciation new photos of human chromosome spreads (Investigation 10 and 11) and the back cover photograph provided by Dr. Daniel Van Dyke and his colleagues at the Mayo Clinic. We thank the Biological Sciences Curriculum Study (Investigation 22), and Helena Laboratories (Investigation 25) for illustrations they provided. The section of Investigation 25 that deals with electrophoresis of hemoglobin was reviewed by Helena Laboratories of Beaumont, Texas, and is used with their permission. Investigation 16 is based in part on material supplied by the Perkin Elmer Corporation and by Carolina Biological Supply Company, and thanks are due to both firms for their assistance. Kits for conducting this investigation may also be purchased from Carolina Biological. We wish to thank our editor Michael Gillespie and assistant editor, Chloé Veylit, and their staff for their support and efforts in the production of this manual. Finally, we wish to thank the instructors and stu-dents who have used the thirteenth edition of this manual. It is their commitment to the manual that has justified the production of the fourteenth edition. We hope that the updates and additions found in the new edition will continue to make the manual a useful instructional tool.

T. R. M. & R. L. H.

in memoriam Thomas Mertens1930–2014Dr. Mertens was a Professor of Biology and Distinguish Professor of Biology Education at Ball State University for over thirty-five years. Among his many accomplishments, he served as president of the National Association of Biology Teachers where he helped guide a strong emphasis on using an “open-ended” approach for teaching genetics, and for which he received their Ohaus Award. In 1988, he received the McGuffey Award from the Textbook and Academic Authors Association for work on this book. Dr. Mertens was the author or co-author of over 200 publications, including this text. He will be greatly missed by his family, colleagues and co-author.

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