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Tested Studies for Laboratory Teaching
Volume 22

Clemson University, June 6-10, 2000

Editor: Susan J. Karcher (Purdue University)
Host: Bob Kosinski

1. Spot-Overlay Ames Test of Potential Mutagens by David R. Wessner¹, Peggy C. Maiorano¹, John Kenyon¹, Ralph Pillsbury², and A. Malcolm Campbell¹ (¹Davidson College and ²Davidson IB Middle School) [abstract] [full text]

2. Using A Molecular Marker to Study Genetic Equilibrium in Drosophila melanogaster by Rodney J. Scott (Wheaton College) [abstract] [full text]

3. Mitochondria and Metabolism in Honeybee Flight Muscle by Alexander F. Motten (Duke University) [abstract] [full text]

4. Mitochondrial DNA from Lumbriculus variegatus: Isolation and Restriction Digest Analysis by Gary Oxford (Longwood College) [abstract] [full text]

5. An Introduction to DNA: Spectrophotometry, Degradation, and the 'Frankengel' Experiment by William Clark and Kimberley Christopher (University of Alberta) [abstract] [full text]

6. DNA Sequencing Used to Illustrate Mutations and Evolution by Theodore Gurney¹, Jr., Russell LeMon¹, and Kathleen Nolan² (¹University of Utah and ²St. Francis College) [abstract] [full text]

7. The Good and the Bad: Symbiotic Organisms from Selected Hosts by Gayle Pittman Noblet and Michael J. Yabsley (Clemson University) [abstract] [full text]

8. Cercariae of Digenetic Trematodes: Use in Laboratory Investigations by Ronald B. Rosen (Berea College) [abstract] [full text]

9. Koch's Crickets: A Study in Etiology by C. Brian Odom (Wingate University) [abstract] [full text]

10. Investigative Laboratories in Cell Biology Using a Host-Parasitoid Model: The Tobacco Hornworm, Manduca sexta, and the Braconid Wasp, Cotesia congregata -- Introduction to the System by Isaure de Buron (Converse College) [abstract] [full text]

11. Plant Reproductive Systems: An Investigative Approach by Laura K. Thompson (Furman University) [abstract] [full text]

12. Why Are Reports of Ant Pollination Rare?: A Field and Lab Exercise Using the Scientific Method by Mary N. Puterbaugh and Mary Blaine Prince (University of Pittsburgh at Bradford) [abstract] [full text]

13. The Line-Intercept Method: A Tool for Introductory Plant Ecology Laboratories by John Cummings and Denny Smith (Clemson University) [abstract] [full text]

14. Birdsong Playback as a Tool for Teaching Animal Behavior by Jerry A. Waldvogel (Clemson University) [abstract] [full text]

15. Murder and Mayhem in Non-Majors Biology by Jean L. Dickey and V. Christine Minor (Clemson University) [abstract] [full text]

16. Bringing the Laboratory into the Lecture Hall by Ruth E. Beattie (University of Kentucky) [abstract] [full text]

17. Effective Methods of Training Biology Laboratory Teaching Assistants II: Preparing TAs To Be Effective in the Laboratory by Jean L. Dickey¹, John Cummings¹, Maggie Haag², Louise McBain², and William Glider³ (¹Clemson University, ² University of Alberta and ²University of Nebraska-Lincoln) [abstract] [full text]

18. Preparation and Presentation of Biology Laboratory Class Materials Through WebCT® by Terry L. Combs (Purdue University) [abstract] [full text]

19. Using Humans as a Central Example in Teaching Undergraduate Biology Labs by Craig E. Nelson¹ and Martin K. Nickels² (¹Indiana University and ²Illinois State University) [abstract] [full text]

Appendix A: Abstracts of Mini Workshops  [titles]

Appendix B: Abstracts of the Workshops presented at the First Regional Association 488 for Biology Laboratory Education (RABLE), Bainbridge College, March 21 - 22, 1996

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Abstracts (Vol. 22)

Laboratory Exercises in Cell and Molecular Biology and Genetics

1 -- Spot-Overlay Ames Test of Potential Mutagens
David R. Wessner, Peggy C. Maiorano, John Kenyon, Ralph Pillsbury, and A. Malcolm Campbell [full text]
Key words: mutagenesis, Salmonella typhimurium, auxotroph, Ames test.
This protocol represents a cost-effective modification of the Ames Test that allows students to investigate the mutagenic potential of various common substances. Potential mutagens are tested using well-characterized auxotrophic strains of Salmonella typhimurium. By analyzing the results, students determine if any of their compounds may be mutagenic. Follow-up experiments are designed to determine the dose response of these potential mutagens. Using this protocol, we have achieved reproducible results with several known mutagens, including sodium azide and ultraviolet irradiation. This approach enables faculty to control costs and results to improve student understanding of mutagenesis, biochemical pathways, experimental design, and data analysis.

2 -- Using A Molecular Marker to Study Genetic Equilibrium in Drosophila melanogaster
Rodney J. Scott [full text]
Key words: Drosophila melanogaster, molecular marker, PCR, Hardy-Weinberg equilibrium.
Using Polymerase Chain Reaction (PCR), genetic variation in a laboratory population of Drosophila is characterized. The population contains flies with two variants of a molecular marker. DNA from individual flies is amplified by PCR, generating products which are either "long" or "short" when visualized on an agarose gel. Three PCR "genotypes" (long/long, long/short, and short/short) are distinguishable and should be present in Hardy-Weinberg frequencies. The exercise requires one session for grinding flies and preparing PCR reactions, and one for conducting and interpreting gel electrophoresis. PCR can be conducted with a standard thermal cycler, or using "hand cycling" with two water baths.

3 -- Mitochondria and Metabolism in Honeybee Flight Muscle
Alexander F. Motten [full text]
Key words: cellular respiration, enzyme, flight muscle, honeybee, mitochondria.
Honeybees provide a convenient source of insect flight muscle, a tissue especially rich in mitochondria. These organelles are easily isolated using simple laboratory equipment and their metabolic activity readily measured colorimetrically with the dye 2,6-dichloroindophenol, which changes from blue to colorless when reduced by the electrons produced during respiration. This system is well-suited for studying cell fractionation, experimental isolation of the major subcellular components responsible for glycolysis and the Krebs cycle, and the role of oxygen in respiration. It can also be used for simple enzymology experiments using the Krebs cycle enzyme succinic dehydrogenase.

4 -- Mitochondrial DNA from Lumbriculus variegatus: Isolation and Restriction Digest Analysis
Gary Oxford [full text]
Key words: mitochondria, mitochondrial DNA, restriction digest, Lumbriculus.
In this activity, mitochondria are isolated from the California blackworm, Lumbriculus variegatus, by homogenization and differential centrifugation. A modified plasmid miniprep procedure (alkaline lysis) is then used to isolate DNA from these mitochondria. Single and double enzyme restriction digests of mitochondrial DNA (mtDNA) are analyzed by agarose gel electrophoresis. Sizes of mtDNA restriction fragments are estimated from gel results and are used to determine the total size of the L. variegatus mitochondrial genome. Issues involved in creating a restriction map of mtDNA from this organism are discussed.

5 -- An Introduction to DNA: Spectrophotometry, Degradation, and the 'Frankengel' Experiment
William Clark and Kimberley Christopher [full text]
Key words: DNA, 'Frankengel', electrophoresis, spectrophotometry.
In this laboratory students perform three exercises as an introduction to the basics of handling and analyzing DNA. In the first, they expose circular plasmid, linear phage, and high molecular weight genomic DNA samples to a variety of physical, thermal, chemical, and enzymatic conditions that might be expected to affect DNA integrity. The DNA's are analyzed by electrophoresis on a group agarose gel. In the second, they pour and reconstruct a "Frankengel" (a gel containing sections with three different agarose concentrations) on which they run a DNA ladder in order to investigate the effect of gel pore size on DNA fragment mobility. In the third, they perform an UV spectrophotometric analysis of DNA from which they learn to make purity and concentration estimates.

6 -- DNA Sequencing Used to Illustrate Mutations and Evolution
Theodore Gurney, Jr., Russell LeMon, and Kathleen Nolan [full text]
Key words: mutation, evolution, PCR, DNA sequencing, systematics, bioinformatics.
This workshop, which is half wet lab and half Internet lab, uses DNA sequences to examine the relatedness of different species. DNA from several insect and/or fish species are compared. This chapter includes instructions for extracting DNA from gels and making PCR products. We prepare PCR products to send to a computerized automated sequencing facility. (No radioisotopes are used in this sequencing!) We recover sequence from the facility over the Web as four-color graphics on a computer screen and as text. We make pair-wise DNA sequence comparisons between species with BLAST2 and multi-species comparisons with MultAlin. We see substitutions, insertions, and deletions. Then we make a distance-based evolutionary tree with GeneBee.

Laboratory Exercises in Ecology, Evolution, and Behavior

7 -- The Good and the Bad: Symbiotic Organisms From Selected Hosts
Gayle Pittman Noblet and Michael J. Yabsley [full text]
Key words: commensals, fish, frogs, fruit flies, mutuals, parasites, pigeons, termites.
Symbiosis is defined as one organism living on, in, or with another organism. Three primary categories are mutualism (both organisms benefit), commensalism (commensal benefits and host not affected), and parasitism (parasite benefits and host harmed). Hands-on dissection of host animals provides examples of live symbiotes in all three types of symbiosis: mutualism: termite flagellates and rumen ciliates; commensalism: Leptomonas flagellates in Drosophila; parasitism: fish hosts (monogeneans on gills, larval digeneans and acanthocephalans in liver) and frogs (trematodes in lungs and bladder; ciliates, flagellates and opalinids in large intestine/cloaca). Possible changes in a relationship from commensalism to parasitism are discussed relative to a demonstration of tapeworm life cycle stages in rat and beetle hosts.

8 -- Cercariae of Digenetic Trematodes: Use In Laboratory Investigations
Ronald B. Rosen [full text]
Key words: cercaria, trematode, Digenea, Proterometra, fish parasites.
In the complex life cycle of a digenetic trematode, the cercaria larva is shed from a snail intermediate host and represents a short-lived, aquatic transfer stage to the next host. Due to its temporary free-living existence, the cercarial stage is ideal for a number of investigative projects. Two general experiments will be conducted with the unusually large cercaria of Proterometra macrostoma. Students assess the effect of: (1) different light wavelengths on the vertical swimming burst distance of this type of cercaria and (2) pH and pepsin on the emergence of the P. macrostoma distome (pre-adult) from its cercarial tail. The latter study represents an in vitro simulation of what naturally occurs in the stomach of the centrarchid fish definitive host.

9 -- Koch's Crickets: A Study in Etiology
C. Brian Odom [full text]
Key words: microbiology, etiology, Koch, cricket, Serratia marcescens.
Students apply Koch's Postulates in an attempt to discover the causative agent of "Pink Plague" a disease that has struck a group of commercially raised crickets. During the course of this exercise the students will isolate the suspected pathogen from one of a group of infected crickets. They will then characterize the isolated organism and use it to infect a new group of crickets to see if they can reproduce the same symptoms in the newly infected group. Reisolation of the suspected pathogen from this second group of organisms will confirm this organism as the etiologic agent of "Pink Plague."

10 -- Investigative Laboratories in Cell Biology Using a Host-Parasitoid Model: The Tobacco Hornworm, Manduca sexta, and the Braconid Wasp, Cotesia congregata -- Introduction to the System
Isaure de Buron [full text]
Key words: cell biology, investigation, parasitoid, tobacco hornworm, braconid wasp.
A series of exercises were developed in the cell biology laboratory using a host-parasite system: the tobacco hornworm and an inoffensive parasitoid wasp. Exercises are fully investigative and no initial schedule is given to the students who, after becoming acquainted with the system, raise questions and decide as a group what they wish to study next (Cell morphology? Cytoskeleton? Protein synthesis?…). This workshop involved introduction to the system of study, phase contrast microscopy, and digital image capturing as examples of how the system can be used. Some of the numerous biological concepts that may be taught using this system are outlined herein. The development of this system was funded by a CCLI-NSF grant (DUE 99-51371).

11 -- Plant Reproductive Systems: An Investigative Approach
Laura K. Thompson [full text]
Key words: plant reproduction, flower structure, pollination, ocular micrometer.
This plant evolution exercise endeavors to move away from a purely observational approach to one of investigation. Students investigate the increasing use of air and animals for gamete and population dispersal that aided plant development on land. The first part of this chapter deals with a common woodland fern of the order Filicales. Students examine sporophyte and gametophyte plants, and experiment with air-borne spore release. The second part of this chapter has two main objectives dealing with the reproductive structures of angiosperms. First, students learn to recognize flower structures and variation in structural patterns. Second, students observe how floral structure and pollen size differ with two agents of pollination, wind and insects.

12 -- Why Are Reports of Ant Pollination Rare?: A Field and Lab Exercise Using the Scientific Method
Mary N. Puterbaugh and Mary Blaine Prince [full text]
Key words: ecology, ants, pollination, mutualism, field study, hypothesis testing.
This exercise uses ants (considered by some to be the "scoundrels in the pollination drama!") to test questions about the morphological and behavioral features that permit insects to be good pollinators. Students are presented with the fact that ants are abundant insects, but fewer than 20 plant species worldwide have been documented as ant-pollinated. Students generate specific hypotheses for the rarity of ant pollination, and design experiments to test their hypotheses. This exercise has both a field and indoor component so that it can be successful even if it is raining, and ants are inactive outdoors.

13 -- The Line-Intercept Method: A Tool for Introductory Plant Ecology Laboratories
John Cummings and Denny Smith [full text]
Key words: sampling techniques, transect sampling, transition zones, plant community structure.
This ABLE major workshop took participants to Bogg's Rock, a granitic flatrock community that is in the early serial stages of succession. The line-intercept method of sampling was employed to allow the rapid characterization of the community structure. After species composition was defined, a simple statistical procedure was used to test one of several factors that might be controlling species distribution.

14 -- Birdsong Playback as a Tool for Teaching Animal Behavior
Jerry A. Waldvogel [full text]
Key words: behavior, birds, birdsong, communication, ecology, field methods.
Animal behavior is often downplayed in general biology labs. Problems include where to deal with the topic in the syllabus, time constraints and special equipment needs, and the unpredictable nature of behavioral responses. Birdsong playback offers a solution to these problems. Birdsong relates well to several important conceptual areas of biology. Its acoustic features can be investigated easily using desktop computers, and it is a fairly reliable behavioral response obtainable in the field. This lab provides general background information about birdsong, explores the structure of birdsong using acoustic analysis computer software, and offers field experience in playback techniques.

Instructional Materials

15 -- Murder and Mayhem in Non-Majors Biology
Jean L. Dickey and V. Christine Minor [full text]
Key words: scientific inquiry, hypothesis testing, non-majors biology, DNA fingerprints.
Who killed James Watson in the biology lab, with the biology textbook? In this non-majors laboratory exercise, students use scientific inquiry skills to solve a murder mystery. Many are suspects, but only one committed the crime. Each student plays a role and tries to uncover motive and opportunity of the other suspects. Hypotheses are tested with physical evidence: fingerprints, blood type, and paper strip DNA analysis.

16 -- Bringing the Laboratory into the Lecture Hall
Ruth E. Beattie [full text]
Key words: exercises in lectures, soil testing, pH, N₂, PD₄, K, water pollution, water quality.
As part of their general education studies, all students at the University of Kentucky must take two natural science courses. Many non-science majors choose the biology sequence of courses. Unfortunately these courses are lecture-only courses, and so some students can graduate without ever having had a science laboratory course. In an effort to provide students with some laboratory experience, I have developed / adapted a number of laboratory activities, which I have successfully incorporated into a non-majors biology course (Human Ecology). These activities are carried out in a lecture hall with 300 students in 50-minute time periods. In this chapter, two of these laboratory activities are presented. 1) Soil Testing - students test soil samples for nitrogen, potassium, phosphate and pH; compare the relative fertility of different soil samples; and carry out an inventory of animal and microbial life in soil samples. 2) Water Pollution Testing - students examine the effects of household chemicals on water quality (from Using Fast Plants and Bottle Biology in the Classroom, published by National Association of Biology Teachers, 1994, ISBN #0-941212-17-3).

17 -- Effective Methods of Training Biology Laboratory Teaching Assistants II: Preparing TAs To Be Effective in the Laboratory
Jean L. Dickey, John Cummings, Maggie Haag, Louise McBain, and William Glider [full text]
Key words: teaching assistants, TA preparation, teaching assessment.
One of the most important factors in the success of undergraduate biology laboratories rests with the instructors, often undergraduate or graduate students, who teach them. This ABLE 2000 workshop, which continued a session presented at ABLE 1999, focused on the theme "teaching effectively in the laboratory." Topics included preparation, organization, supervision of group work, and involvement of students in discussion. In addition to presentations by the workshop organizers, participants engaged in lively discussion of their own experiences and methods.

18 -- Preparation and Presentation of Biology Laboratory Class Materials Through WebCT®
Terry L. Combs [full text]
Key words: WebCT©, course, online, pdf, jpeg, gif.
An introduction to optimizing materials for web presentation: postscripting to PDF, animated graphics, still graphics, font type and size. These materials, among others, are used to demonstrate the creation of a WebCT© course site. WebCT© is a server-based course management software application. Through a very specific (and thoroughly, yet simply explained) set of steps, faculty place html, text files, graphics files, etc. into what is essentially an on-line course. That 'course' allows you to place your study materials and outlines on the web, create self-testing and scoring for individual materials, give quizzes and exams, search your own glossary, keep an on-line calendar that links to files or sites inside or outside your course, keep track of grades, assign each of your students email capability, run chat rooms, publish a bulletin board, and allow you to track students' study time and materials. All creation work and student participation can be done wherever there is a browser and WWW access.

19 -- Using Humans as a Central Example in Teaching Undergraduate Biology Labs
Craig E. Nelson and Martin K. Nickels [full text]
Key words: evolution, taxonomy, human, fossil hominoids, primates, skull, molecular sequences, hemoglobin.
Focusing on human and primate examples significantly enhances students' interest in studying biology, specifically, evolution and classification. These laboratory activities illustrate key aspects of the distinctive hierarchical nature of biological classification. The first activity uses hemoglobin sequence data while the second uses skulls of apes, humans and fossil hominoids. Together, the two illustrate the concordant nature of separate lines of evidence supporting both the idea of evolution generally and that of human evolution specifically.

Mini Workshops

• DNA: A Kinesthetic Experience by Sandra Johnson (Middle Tennessee State University) [full text]
• Dispersal Cues Used by Parasitic Wasps: Phototaxis, Geotaxis and Mated Melittobia by Judith A. Guinan¹ and Robert W. Matthews² (¹Hollins University and ²University of Georgia) [full text]
• Tackling Phylogeny in Introductory Biology Laboratories by Marianne Niedzlek-Feaver¹ and John W. Zimmerman² (¹North Carolina State University and ²Mount St. Clair College) [full text]
• Rearing Xenopus laevis Life History Stages by Ron Koss and Bruce Wakeford (University of Alberta) [full text]
• Metabolism and Oxygen Consumption in Aquatic Organisms by Ruthanne B. Pitkin (Shippensburg University) [full text]
• Investigating Your Watershed: Detection of Fecal Coliform Bacteria by Karin Readel (University of Maryland, Baltimore County) [full text]
• A Simple Sampling Method That Lets Students See the Forest AND the Trees by Janet W. Phelps (University of Wisconsin - Baraboo/Sauk County) [full text]
• Use of Video Microscopy to Promote Collaborative Learning in Developmental Anatomy and Histology by Betty L. Black and Harold G. Meeks, Jr. (North Carolina State University) [full text]
• Using Projects to Encourage Exploration of Animal Diversity and Educational Resources by Jean L. Dickey and Jeanne Leonard (Clemson University) [full text]
• Introducing Inquiry In A Biology Lab For Non-Majors by Phyllis S. Laine and Linda J. Heath (Xavier University) [full text]
• A Computer-Based Approach for Generating Standardized Student Assessment Using a First Year Biology Programme as a Model to Evaluate Its Effectiveness by Todd Nickle and Jeffrey Sheedy (Mount Royal College) [full text]
• Team Projects: The Good, The Bad, and the Ugly by Joy B. Perry (University of Wisconsin - Fox Valley) [full text]
• On-Line Manual Enhances Student Learning by Arthur L. Buikema, Jr., William Zabaronick, and Mary Alice Schaeffer (Virginia Polytechnic Institute and State University) [full text]
• The Use of Interactive Web-Based Courseware in Intro Biology by Corey A. Goldman (University of Toronto) [full text]
• Developing Biology Board Games by William Beachy (Hastings College) [full text]
• Interactive Exploration of Kingdom Animalia Using the Animal Diversity Web by Cynthia Sims Parr (University of Michigan) [full text]
• Using Student-Centered Investigative Modules to Teach a Multi-Disciplinary Scientific Methods Laboratory Course to Undergraduate Non-Science Major Students by John D. Usis (Youngstown State University) [full text]
• Connecting Teaching Lab Development and Education Research: An Introduction by Brian White (University of Massachusetts - Boston) [full text]
• Student Reviews of Scientific Literature: Opportunities to Improve Students' Scientific Literacy and Writing Skills by Susan J. Karcher (Purdue University) [full text]