Biology Laboratory Manual Twelfth Edition Darrell S. Vodopich Baylor University Randy Moore University of Minnesota vod00720_fm_i-xii.indd 1 10/16/18 1:57 PM BIOLOGY LABORATORY MANUAL, TWELFTH EDITION Published by McGraw-Hill Education, 2 Penn Plaza, New York, NY 10121. Copyright © 2020 by McGraw-Hill Education. All rights reserved. Printed in the United States of America. Previous editions © 2017, 2014, and 2011. No part of this publication may be reproduced or distributed in any form or by any means, or stored in a database or retrieval system, without the prior written consent of McGraw-Hill Education, including, but not limited to, in any network or other electronic storage or transmission, or broadcast for distance learning. Some ancillaries, including electronic and print components, may not be available to customers outside the United States. This book is printed on acid-free paper. 1 2 3 4 5 6 7 8 9 LMN 21 20 19 ISBN 978-1-260-20072-0 (bound edition) MHID 1-260-20072-8 (bound edition) ISBN 978-1-260-41330-4 (loose-leaf edition) MHID 1-260-41330-6 (loose-leaf edition) Portfolio Manager: Andrew Urban Product Developer: Donna Nemmers Marketing Manager: Kelly Brown Content Project Managers: Jessica Portz & Sandra Schnee Buyer: Laura Fuller Design: David W. Hash Content Licensing Specialist: Lorraine Buczek Cover Image: ©Darrell S. Vodopich Compositor: MPS Limited All credits appearing on page are considered to be an extension of the copyright page. Some of the laboratory experiments included in this text may be hazardous if materials are handled improperly or if procedures are conducted incorrectly. Safety precautions are necessary when you are working with chemicals, glass test tubes, hot water baths, sharp instruments, and the like, or for any procedures that generally require caution. Your school may have set regulations regarding safety procedures that your instructor will explain to you. Should you have any problems with materials or procedures, please ask your instructor for help. The Internet addresses listed in the text were accurate at the time of publication. The inclusion of a website does not indicate an endorsement by the authors or McGraw-Hill Education, and McGraw-Hill Education does not guarantee the accuracy of the information presented at these sites. mheducation.com/highered vod00720_fm_i-xii.indd 2 10/16/18 1:57 PM CCoonntteennttss Preface v Exercise 16 Teaching and Learning Tools viii Molecular Biology and Biotechnology: DNA Isolation Welcome to the Biology Laboratory ix and Genetic Transformation 171 Exercise 17 Exercise 1 Genetics: The Principles of Mendel 179 Scientific Method: The Process of Science 1 Exercise 18 Exercise 2 Evolution: Natural Selection and Morphological Change Measurements in Biology: The Metric System and Data Analysis 11 in Green Algae 195 Exercise 3 Exercise 19 The Microscope: Basic Skills of Light Microscopy 21 Human Evolution: Skull Examination 207 Exercise 4 Exercise 20 The Cell: Structure and Function 33 Ecology: Diversity and Interaction in Plant Communities 217 Exercise 5 Exercise 21 Solutions, Acids, and Bases: The pH Scale 49 Community Succession 227 Exercise 6 Exercise 22 Biologically Important Molecules: Carbohydrates, Proteins, Lipids, Population Growth: Limitations of the Environment 235 and Nucleic Acids 57 Exercise 23 Exercise 7 Pollution: The Effects of Chemical, Thermal, Separating Organic Compounds: Column Chromatography, and Acidic Pollution 243 Paper Chromatography, and Gel Electrophoresis 71 Exercise 24 Exercise 8 Survey of Prokaryotes: Domains Archaea and Bacteria 253 Spectrophotometry: Identifying Solutes and Determining Their Concentration 81 Exercise 25 Survey of Protists: The Algae 269 Exercise 9 Diffusion and Osmosis: Passive Movement of Molecules Exercise 26 in Biological Systems 93 Survey of Protists: Protozoa and Slime Molds 283 Exercise 10 Exercise 27 Cellular Membranes: Effects of Physical and Chemical Stress 105 Survey of the Kingdom Fungi: Molds, Sac Fungi, Mushrooms, and Lichens 293 Exercise 11 Enzymes: Factors Affecting the Rate of Activity 113 Exercise 28 Survey of the Plant Kingdom: Liverworts, Mosses, and Hornworts Exercise 12 of Phyla Hepaticophyta, Bryophyta, and Anthocerophyta 307 Respiration: Aerobic and Anaerobic Oxidation of Organic Molecules 125 Exercise 29 Exercise 13 Survey of the Plant Kingdom: Seedless Vascular Plants of Phyla Photosynthesis: Pigment Separation, Starch Production, Pterophyta and Lycophyta 317 and CO Uptake 137 2 Exercise 30 Exercise 14 Survey of the Plant Kingdom: Gymnosperms of Phyla Cycadophyta, Mitosis: Replication of Eukaryotic Cells 149 Ginkgophyta, Coniferophyta, and Gnetophyta 329 Exercise 15 Exercise 31 Meiosis: Reduction Division and Gametogenesis 159 Survey of the Plant Kingdom: Angiosperms 339 TOC–1 iii vod00720_fm_i-xii.indd 3 10/16/18 1:57 PM Exercise 32 Exercise 43 Plant Anatomy: Vegetative Structure of Vascular Plants 355 Human Biology: Muscles and Muscle Contraction 497 Exercise 33 Exercise 44 Plant Physiology: Transpiration 369 Human Biology: Breathing 505 Exercise 34 Exercise 45 Plant Physiology: Tropisms, Nutrition, and Growth Regulators 377 Human Biology: Circulation and Blood Pressure 515 Exercise 35 Exercise 46 Bioassay: Measuring Physiologically Active Substances 389 Human Biology: Sensory Perception 529 Exercise 36 Exercise 47 Survey of the Animal Kingdom: Phyla Porifera and Cnidaria 395 Vertebrate Anatomy: External Features and Skeletal System of the Rat 539 Exercise 37 Survey of the Animal Kingdom: Phyla Platyhelminthes Exercise 48 and Nematoda 411 Vertebrate Anatomy: Muscles and Internal Organs of the Rat 547 Exercise 38 Exercise 49 Survey of the Animal Kingdom: Phyla Mollusca and Annelida 425 Vertebrate Anatomy: Urogenital and Circulatory Systems of the Rat 557 Exercise 39 Exercise 50 Survey of the Animal Kingdom: Phylum Arthropoda 439 Embryology: Comparative Morphologies and Strategies of Development 569 Exercise 40 Survey of the Animal Kingdom: Phyla Echinodermata and Chordata 453 Exercise 51 Animal Behavior: Taxis, Kinesis, and Agonistic Behavior 579 Exercise 41 Vertebrate Animal Tissues: Epithelial, Connective, Muscular, Appendix I and Nervous Tissues 473 Dissection of a Fetal Pig 585 Exercise 42 Appendix II Human Biology: The Human Skeletal System 489 Conversion of Metric Units to English Units 592 iv TOC–2 vod00720_fm_i-xii.indd 4 10/16/18 1:57 PM CProenftaecnets W e have designed this laboratory manual for an intro- discussions and interactions between student and instructor ductory biology course with a broad survey of basic are major components of a successful laboratory experi- laboratory techniques. The experiments and procedures are ence. Be sure to examine the “Questions for Further Study simple, safe, easy to perform, and especially appropriate for and Inquiry” in each exercise. We hope they will help you large classes. Few experiments require more than one class expand students’ perceptions that each exercise has broad meeting to complete the procedure. Each exercise includes application to their world. many photographs and illustrations, traditional topics, and experiments that help students learn about life. Procedures DIGITAL INTEGRATION within each exercise are numerous and discrete so that an exercise can be tailored to the needs of the students, the style As educators, we recognize that today’s students are digital of the instructor, and the facilities available. learners. Virtually every exercise of this manual is accom- panied by tailor-made digital resources, including assign- TO THE STUDENT able questions and a variety of high-definition videos, PowerPoint images, and other resources that demonstrate We hope this manual is an interesting guide to many areas basic techniques, emphasize biological principles, test for of biology. As you read about these areas, you’ll probably understanding, and engage students as they learn biology spend equal amounts of time observing and experimenting. in the laboratory. Don’t hesitate to go beyond the observations that we’ve Digital resources are available to instructors at connect outlined—your future success as a scientist and an informed .mheducation.com. Instructors will want to assign these citizen depends on your ability to seek and notice things that resources to help students know what they’ll be doing, what others may overlook. Now is the time to develop this ability principles they’ll be investigating, and what concepts they’ll with a mixture of hard work and relaxed observation. Have need to understand before coming to lab. fun, and learning will come easily. Also, remember that this manual is designed with your instructors in mind as well. Go to them often with questions—their experience is a valuable WHAT’S NEW IN THIS EDITION tool that you should use as you work. Throughout the manual, we have expanded and improved several of the most popular and effective features of TO THE INSTRUCTOR previous editions, including This manual’s straightforward approach emphasizes experi- ∙ Learning Objectives have been updated to provide ments and activities that optimize students’ investment of an overview of what students will do and learn in the time and your investment of supplies, equipment, and prepa- exercise. ration. Simple, safe, and straightforward experiments are most effective if you interpret the work in depth. Most exper- ∙ Procedures and Doing Biology Yourself require stu- iments can be done easily by a student in 2 to 3 hours. Ter- dents to do biology as they apply skills they’ve learned minology, structures, photographs, and concepts are limited to develop and study hypotheses about biology. to those that the student can readily observe and understand. ∙ Questions throughout each exercise encourage students In each exercise we have included a few activities requiring to pause and think about their data and what they’ve a greater investment of effort if resources are available, but learned in lab. omitting them will not detract from the objectives. ∙ Questions for Further Study and Inquiry at the This manual functions best with an instructor’s guid- end of each exercise help students apply what they’ve ance and is not an autotutorial system. We’ve tried to guide learned to broader topics and issues in biology. students from observations to conclusions, to help students make their own discoveries, and to make the transition from ∙ Writing to Learn Biology encourages students to observation to understanding biological principles. But develop their ideas about what they learned in lab. P–1 v vod00720_fm_i-xii.indd 5 10/16/18 1:57 PM ∙ Caution and Safety First icons make students aware of ∙ Exercise 13—Figure 13.2 caption is expanded safety issues associated with the procedures they’ll use ∙ Exercise 14—Explanation of the structure of chromatids in lab. is expanded ∙ Boxed readings titled Inquiry-Based Learning encourage ∙ Exercise 15—Labels for figure 15.2 have been added students to apply what they’ve learned to independently for paternal versus maternal chromosomes; description answer questions about intriguing biological topics. of the structure of replicated versus nonreplicated ∙ Updated health-related exercises help students better chromosomes has been clarified; figure 15.6 is new; understand topics such as blood pressure, atherosclerosis, figure 15.7 is revised to clarify the state and number and their risk of cardiovascular disease. of chromosomes in first polar bodies and second polar bodies, and corpus albicans has been labeled and added ∙ Several illustrations have been replaced with photographs as a defined term in the text to provide more realistic images to support the Exercise content. ∙ Exercise 16—Global prevalence of genetically trans- formed crops has been updated to 2017 statistics ∙ Approximately 60 illustrations and photos have been revised. ∙ Exercise 17—Figure 17.4 has a panel of 3 new photos on sickle cell anemia; figure 17.6 contains improved ∙ Questions within procedures now include lines on which photos of hairlines students can write their answers. ∙ Exercise 18—Definition of evolution is revised to ∙ An assignable, updated library of videos and Connect be more concise; questions about Hardy-Weinberg questions helps students prepare for lab and understand genetics are expanded for clarity; a new question the instruments and techniques that will be important about the effect of natural selection on sickle cell for their investigations. Instructors may assign these vid- anemia has been added to Questions for Further Study eos before class time to help ensure that students arrive and Inquiry prepared for lab. ∙ Exercise 19—Figure 19.2 has been revised to better illustrate lineages of human evolution; the term Exercise-Specific Changes “diastema” has been added and defined; figure 19.4 ∙ Exercise 1—Additional explanation provided for both is relabeled for clarity mean and standard deviation ∙ Exercise 20—Procedure 20.4 is expanded to help ∙ Exercise 2—Mass, volume, and median are further students design and implement experimental controls. defined; new illustration in figure 2.3 on measuring the ∙ Exercise 22—Formula for population growth is revised; volume of liquid; figure 2.4b has explanatory labels data for Figure 22.5 are updated to reflect 2018 predic- added tions; question 6 is expanded to include 2018 population ∙ Exercise 3—Additional questions have been added to values and growth rates Procedure 3.6 Using a dissecting microscope ∙ Exercise 23—Question 1 is revised to emphasize ∙ Exercise 4—Several illustrations have better labels; a new hypothesis testing; table 23.3 is reorganized to accept photo is supplied for figure 4.6a Elodea cells; figure 4.13 handwritten student data has been redrawn to more directly correlate to the associ- ∙ Exercise 24—Organization of domains and kingdoms ated photo; a new question is added to Questions for Fur- is updated to current taxonomy; table 24.1, prokaryotic ther Study and Inquiry to compare plant and animal cells versus eukaryotic characteristics, is modified for preci- ∙ Exercise 6—Qualitative tests are defined; a new photo sion; figure 24.2, structure of a bacterial cell, is revised has been added to figure 6.2 to explain Benedict’s test and contains a new photo; explanation of binary fission is expanded to include protein FtsZ and its role in cell ∙ Exercise 7—Clarifying edits made to introductory separation material ∙ Exercise 25—Explanations of Archaeplastida and the ∙ Exercise 9—Explanations of hypotonic, hypertonic, and term “protist” are clarified; in table 25.2 the list of isotonic are expanded chlorophylls diagnostic to each type of algae is updated; ∙ Exercise 10—Steps of Procedures 10.1 and 10.2 are figure 25.4 is relabeled to clarify sexual versus asexual clarified; a new question on experimental design has reproductive paths; figure 25.8 contains a new photo of been added to Questions for Further Study and Inquiry Volvox colonies vi P–2 vod00720_fm_i-xii.indd 6 10/16/18 1:57 PM ∙ Exercise 26—Photomicrograph and illustration of conventional values; figure 42.2 is new; Figure 42.4 African sleeping sickness blood cells and parasites are has been replaced with improved images of normal and revised to clarify their relationship osteoporotic bone; revisions to Questions for Further Study and Inquiry ∙ Exercise 27—Explanations of fungal sporangiophores and sporangia are expanded; figure 27.13 is modified ∙ Exercise 43—A new learning objective is added to to better show the diagnostic reproductive structure, distinguish between isotonic and isometric contractions; ascus; Questions for Further Study and Inquiry has a explanations of muscle load, muscle tone, and muscle new question to explain the benefit of fungi to other tension are expanded; figure 43.2 is relabeled to clearly organisms distinguish between flexion and extension; Procedure 43.1 concerning flexion and extension of the forearm ∙ Exercise 31—A learning objective is added on under- has been modified for clarity standing flower structure and function; the explanation of sporogenesis is expanded; a Question for Further ∙ Exercise 44—Descriptions of negative pressure and its Study and Inquiry has been added to help students role in breathing have been expanded; procedures to dis- understand flower parts tinguish the role of intercostal muscles and breathing are expanded and clarified; Procedure 44.1 has been modi- ∙ Exercise 32—A new question is added to Questions fied for more consistent chest expansion measurements; for Further Study and Inquiry on common leaf typical values for tidal, expiratory, inspiratory, and morphologies residual volumes have been provided; directions for ∙ Exercise 35—The definition of bioassay is revised measuring breathing rate in Procedure 44.7 are clarified ∙ Exercise 36—Introductions to terms animals, multi- ∙ Exercise 46—Figure 46.1 has been modified to illustrate cellular, ancient, and primitive have been clarified; fovea centralis; Procedure 46.3 has been modified to description of intracellular versus extracellular digestion accommodate lab partners in poriferans has been clarified ∙ Exercise 47—A new Question 2 has been added; ∙ Exercise 37—Taxonomic hierarchy of the classes and Question 3 has been expanded to provide more subphyla of flatworms is updated; the groups Neoder- examples and practice with terms such as cranial, mata and Turbellaria have been redefined and updated; caudal, lateral, distal, etc.; directions for the taxonomy of tapeworms is updated skinning and abdominal incision during rat dissection ∙ Exercise 39—Taxonomy of major arthropod classes has are expanded been updated and reorganized to include Chelicerata, ∙ Exercise 48—Descriptions of the thyroid gland and Crustacea, Myriapoda, and Hexapoda; table 39.3 has diaphragm are expanded; explanatory questions about been relabeled to reflect updated arthropod taxonomy the lung structure and heart musculature are expanded ∙ Exercise 40—The taxonomy of pre-vertebrate groups ∙ Exercise 49—Figure 49.4 has been revised and enlarged has been updated; class Actinopterygidii has replaced to better show the structure and cross section of a Osteichthyes; figure 40.21 of amphibian transitional kidney stages is revised ∙ Exercise 50—Distinction has been enhanced between ∙ Exercise 41—Figure 41.2 has revised labeling; the animal and vegetal poles figure 41.3 is relabeled to distinguish flat cuboidal and ∙ Exercise 51—Directions are enhanced for Procedure columnar cells more clearly; figure 41.4 is relabeled 51.1 to examine kinesis in pill bugs; directions are to show Bowman’s capsule more clearly; figure 41.5 enhanced for Procedure 51.2 to study agonistic is relabeled to more clearly distinguish columnar cells; behavior in fighting fish, to encourage better creativity figure 41.7 has been replaced to better show stratified by the students in experimental design; a new question squamous epithelium; types of connective tissue have has been added to Questions for Further Study and been separated into connective tissue proper and special Inquiry connective tissue ∙ Appendix II has been updated to include upcoming ∙ Exercise 42—Descriptions of the appendicular skel- changes to how a basic unit of the metric system is eton and the axial skeleton are added; the number of defined skull, spine, and rib cage bones has been updated to P–3 vii vod00720_fm_i-xii.indd 7 10/16/18 1:57 PM TeCaocnhtinengt asnd Learning Tools McGraw-Hill Connect® Biology syllabus or teaching notes. Find the content you need in Create by searching through thousands of leading McGraw-Hill text- McGraw-Hill Connect Biology provides online presenta- books. Arrange your book to fit your teaching style. Create tion, assignment, and assessment solutions. It connects your even allows you to personalize your book’s appearance students with the tools and resources they’ll need to succeed by selecting the cover and adding your name, school, and at connect.mheducation.com. course information. Order a Create book and you’ll receive a complimentary print review copy in 3–5 business days or a With Connect Biology, you can deliver assignments and complimentary electronic review copy (eComp) via e-mail in quizzes online. A robust set of questions and activities is minutes. Go to create.mheducation.com today and register presented and aligned with this lab manual’s learning out- to experience how McGraw-Hill Create empowers you to comes. Pre-lab worksheets and Investigation worksheets teach your students your way. are also included within Connect. As an instructor, you can edit existing questions and write entirely new questions. Laboratory Resource Guide Track students’ performance—by question, by assignment, or in relation to the class overall—with detailed grade reports. The Laboratory Resource Guide is essential for instructors Integrate grade reports easily with Learning Management and laboratory assistants and is available free to adopters of Systems (LMS), such as Blackboard—and much more. the Laboratory Manual within Connect under the Instructor Resources tab. McGraw-Hill CreateTM With McGraw-Hill Create, you can easily rearrange exer- cises, combine material from other content sources, and quickly upload content you have written, such as your course viii T–1 vod00720_fm_i-xii.indd 8 10/16/18 1:57 PM CWoenlcteonmtes to the Biology Laboratory Welcome to the biology laboratory! Although reading own experiments to answer questions that you’ve posed. your textbook and attending lectures are important To make these exercises most useful and enjoyable, follow ways of learning about biology, nothing can replace the these guidelines noted in the next sections. importance of the laboratory. In lab you’ll get hands-on experience with what you’ve heard and read about biology— for example, you’ll observe organisms, do experiments, test THE IMPORTANCE OF COMING TO CLASS ideas, collect data, and make conclusions about what you’ve learned. You’ll do biology. Biology labs are designed to help you experience biology You’ll enjoy the exercises in this manual—they’re firsthand. To do well in your biology course, you’ll need to interesting and informative and can be completed within attend class and pay attention. To appreciate the importance the time limits of your laboratory period. We’ve provided of class attendance as it relates to making a good grade in questions to test your understanding of what you’ve done; in your biology course, examine figure 1, which is a graph some of the exercises, we’ve also asked you to devise your showing how students’ grades in an introductory biology 100 A B 80 C D 60 %) e ( d a Gr 40 F 20 0 0 20 40 60 80 100 Attendance (% of classes attended) Figure 1 Relationship of students’ grades in an introductory biology course to their rates of class attendance. W–1 ix vod00720_fm_i-xii.indd 9 10/16/18 1:57 PM