Home > Contents of Proceedings > Volume 19

Tested Studies for Laboratory Teaching
Volume 19

The University of Calgary, June 10-14, 1997

[Out of print]
Editor: Susan J. Karcher (Purdue University)
Chair of Host Committee: Charles (Chuck) Curry

2. Measuring Genetic Variability in Natural Populations by Allozyme Electrophoresis by James M. Bader (Department of Biology, Case Western Reserve University) [abstract] [full text]

3. Nonradioactive Screening of an M13 Phage Library for An Eucaryotic Microsatellite DNA Sequence by William Clark and Curtis Strobeck (Department of Biological Sciences, University of Alberta) [abstract] [full text]

4. Patching and Capping: Plasma Membrane Protein Redistribution by Warren Gallin and Kimberley Christopher (Department of Biological Sciences, University of Alberta) [abstract] [full text]

5. Signal Transduction and the Control of the Cell Cycle in Yeast (Saccharomyces cerevisiae): A Collaborative Laboratory Exercise by Barbara Hoopes, Nancy Pruitt, Kathleen Baier, and Sherry Brooks (Biology Department, Colgate University) [abstract] [full text]

6. Enzyme Kinetics by Vicki Chandler, Sam Donovan, Will Goodwin, Karen Sprague, and Carl A. Stiefbold (Department of Biology, University of Oregon) [abstract] [full text]

7. Plant Transformation by Lux+ Agrobacterium by Anna Szenthe and William J. Page (Department of Biological Sciences, University of Alberta) [abstract] [full text]

8. BioLab: Using Yeast Fermentation as a Model for the Scientific Method by Helen K. Pigage, Lt. Col. Milton C. Neilsen and Michele M. Greeder (Department of Biology, United States Air Force Academy, Colorado) [abstract] [full text]

9. A Beginner's Guide to the Study of Plant Structure by Edward C. Yeung (Department of Biological Sciences, University of Calgary) [abstract] [full text]

10. Laboratory Investigations With C-Fern^TM (Ceratopteris richardii) by Leslie G. Hickok and Thomas R. Warne (Department of Botany, University of Tennessee) [abstract] [full text]

11. Pungency Assessment in Onions by Catherine A. Teare Ketter (Department of Biological Sciences, University of Georgia) and William M. Randle (Department of Horticulture, University of Georgia) [abstract] [full text]

12. Sampling, Density Estimation and Spatial Relations by Maggie Haag and William M. Tonn (Department of Biological Sciences, University of Alberta) [abstract] [full text]

13. What Can be Learned about Forest Dynamics from the Age Distribution of Trees? by Sheri L. Gutsell and Edward A. Johnson (Department of Biological Sciences, University of Calgary) [abstract] [full text]

14. Forest Fragmentation in the Rocky Mountains by Rebecca Reed (Department of Botany, University of Wyoming), Jane Beiswenger (Department of Zoology and Physiology, UW), and Julia Johnson-Barnard (Department of Rangeland Ecology and Watershed Management, UW) [abstract] [full text]

15. Variability and Selection in Natural Populations of Wood Lice by Rudi C. Berkelhamer (Department of Ecology and Evolutionary Biology, University of California, Irvine) [abstract] [full text]

16. Factors Affecting the Spread of Disease in Human Populations by Nancy L. Goodyear (Division of Sciences, Bainbridge College) [abstract] [full text]

17. The Mammalian Masticatory Apparatus: An Introductory Comparative Exercise by Anthony P. Russell (Department of Biological Sciences, University of Calgary) [abstract] [full text]

18. Use of Impedance Pneumography in Exercise Physiology by Connie Brewer and Mary Gray, (Department of Biology, Purdue University) [abstract] [full text]

19. Developing Course and Laboratory Homepages for the World Wide Web by Glenn A. Bauer (Biology Department, Saint Michael's College) and Glenn MacDougall (Acadia Development Centre, Acadia University) [abstract] [full text]

20. Enhancing Your Biology Course and Lab WWW Pages with the Latest in HTML, CGI, and JavaScript Features by Glenn MacDougall (Acadia Development Centre, Acadia University) and Glenn A. Bauer (Biology Department, Saint Michael's College) [abstract] [full text]

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Abstracts (Vol. 19)

Laboratory Exercises in Cell and Molecular Biology and Genetics

1 -- Bacterial Gene Transfer
John Mordacq and Roberta Ellington [full text]
Key Words: conjugation, auxotroph, Escherichia coli, chromosome, mapping, recombination.
This laboratory exercise investigates one form of genetic recombination in bacteria. This process, called conjugation, occurs when one bacteria transfers DNA to another bacteria. Two different strains of Escherichia coli are used: an Hfr (high frequency of recombination) strain with the F (fertility) factor integrated into the bacterial chromosome acting as the donor, and an F^- strain lacking the fertility factor acting as a recipient. The F^- strain is auxotrophic for certain genetic markers and the ordered transfer of markers from the Hfr strain to the F^- strain is used to map gene locations on the bacterial chromosome.

2 -- Measuring Genetic Variability in Natural Populations by Allozyme Electrophoresis James M. Bader [full text]
Key Words: allozymes, cellulose acetate electrophoresis, genetic variability, genetic distance.
This laboratory investigates the genetic variability in three populations of bark beetles (Scolytidae). Cellulose acetate electrophoresis is used as a rapid procedure for the separation of allozymes from whole organisms or tissue homogenates, allowing the screening of hundreds of individuals in a three hour laboratory period. Interpretation of gels is straightforward for many enzymes, and calculation of intra- and interpopulation variability is accomplished with a standard spreadsheet program. From this data, inferences can be made regarding the relatedness of the populations and the extent of gene flow between them.
[Also see Measuring Genetic Variation in Zebra Mussels Using Protein Electrophoresis]

3 -- Nonradioactive Screening of an M13 Phage Library for An Eucaryotic Microsatellite DNA Sequence
William Clark and Curtis Strobeck [full text]
Key Words: nonradioactive, biotinylated DNA probes, primary library screen, M13 bacteriophage clones, plaque lifts, hybridization, microsatellites.
The object of this laboratory is to introduce students to a simple, nonradioactive method for the screening of a DNA library for a particular DNA sequence. The library they screen is produced in a bacteriophage M13 cloning vector. The library has been plated out in advance. Students perform a primary screen for microsatellite sequences on plaque lifts hybridized to a nonradioactive (biotinylated) DNA probe. They apply a colorimetric detection system that takes advantage of the high affinity of alkaline phosphatase-conjugated streptavidin for biotin. Blue positives appear amongst a background of clears. By using small, "random" mammalian genomic DNA fragments for the cloning, and probing with a sequence complementary to a common microsatellite repeated sequence, the recovery of at least some positive clones by the students is ensured, and we are able to demonstrate the power of the screening procedure.

4 -- Patching and Capping: Plasma Membrane Protein Redistribution
Warren Gallin and Kimberley Christopher [full text]
Key Words: immunofluorescence, fluid mosaic, membrane-cytoskeleton interaction, membrane fluidity, lymphocyte.
This exercise provides students with practical experience in cell manipulation and immunofluorescent staining, and applies the Fluid Mosaic model of membrane structure. It is based on work by Taylor et al. (1971. Nature New Biology 233: 225-229). Mouse spleen cells are approximately 40% B cells, which express membrane-associated IgM. When this IgM is cross-linked by fluorescein isothiocyanate labeled anti-mouse (FITC-anti-mouse) Ig, the crosslinked proteins form small patches, independent of metabolic energy. Then the patches are collected into a cap at one end of the cell in an energy-dependent process. The FITC on the cross-linking antibody allows one to visualize the process using a fluorescence microscope.

5 -- Signal Transduction and the Control of the Cell Cycle in Yeast (Saccharomyces cerevisiae)
Barbara Hoopes, Nancy Pruitt, Kathleen Baier, and Sherry Brooks [full text]
Key Words: a factor, cell cycle, signal transduction, Saccharomyces cerevisiae, yeast.
This investigative laboratory exposes students to signal transduction. Haploid yeast cells produce a mating factor that activates a signal transduction pathway. Activation results in cell cycle arrest and increased transcription of genes involved in mating. Yeast cells containing mutations in two portions of the signal transduction pathway are transformed with a plasmid coupling b-galactosidase activity to transcription of a mating gene. Cells are assayed with and without mating factor for increased transcription and cell cycle arrest. The effect of mating factor on these two processes allows the determination of what steps in signal transduction are affected by the mutations.

6 -- Enzyme Kinetics
Vicki Chandler, Sam Donovan, Will Goodwin, Karen Sprague, and Carl A. Stiefbold [full text]
Key Words: enzyme kinetics, Km, Vmax, b-galactosidase, ONPG.
The objectives of this laboratory exercise are to understand the principles of an enzyme assay using a spectrophotometer, to understand why the rate of an enzyme reaction reaches a maximum velocity (Vmax) and to understand how the affinity of an enzyme for its substrate (Km) is measured. Students first view computer animation software depicting generalized enzyme catalysis. This is followed by an introduction to absorption spectroscopy and the enzyme assay for b-galactosidase using ONPG as a substrate. Students then estimate Km for b-galactosidase by measuring reaction rates at four substrate concentrations. Lastly, using another computer application, students measure Km and Vmax for wild type -galactosidase enzyme and by measuring Km and Vmax of mutant enzymes determine whether the mutations are in the substrate binding site or catalytic site of the mutant enzymes.

7 -- Plant Transformation by Lux+ Agrobacterium
Anna Szenthe and William J. Page [full text]
Key Words: Agrobacterium, plant transformation, Lux gene.
This laboratory exercise studies the basic biotechnology technique of Agrobacterium-mediated in vivo transformation of N. tabacum cv. SR1 leaf explant with plant expression vector containing an auxin-regulated promoter fused to bacterial luciferase marker gene. Leaf explants will be regenerated on callus generation medium containing auxin, cytokinin and selection for recombinant transgenic callus formation. After determining the soluble cell protein content of callus by Bradford Dye-Binding protein assay, regenerated calli will be assayed for luciferase gene expression by a luminescence photometer and the specific activity will be expressed by mv per minute per mg protein.

8 -- BioLab: Using Yeast Fermentation as a Model for the Scientific Method
Helen K. Pigage, Lt. Col. Milton C. Neilsen and Michele M. Greeder [full text]
Key Words: fermentation, yeast, scientific method.
The BioLab computer simulation accompanies a two-part laboratory exercise with four basic goals: a) student application of the scientific method, experimentation using yeast cells and simple respirometers in the laboratory setting, and repetition to validate laboratory techniques; b) reinforcement of subject matter; c) student writing of a scientific paper with instructor input and revision by the students, and d) student comparison, analysis and synthesis of results from several parts of the laboratory experiment and computer simulation. BioLab addresses and reinforces several specific learning outcomes for the introductory level biology course for majors and nonmajors at the United States Air Force Academy.

Laboratory Exercises in Botany and Plant Development

9 -- A Beginner's Guide to the Study of Plant Structure
Edward C. Yeung [full text]
Key words: free hand sectioning, maceration, plant anatomy, staining.
An understanding of the basic construction of the plant body is essential to the study of plant biology. The purpose of these laboratory exercises is to demonstrate a variety of simple methods that allow students to study plant structures. The following procedures will be introduced to the participants: (1) Free hand sectioning procedures; (2) Histological and histochemical staining of fresh hand sections; (3) Maceration of specimens to show features of plant cells; and (4) Autofluorescence characteristics of plant cells and tissues for the purpose of identification. All these are low cost exercises in which the structural organization of plant cells, tissues, and organs can readily be studied.

10 -- Laboratory Investigations With C-Fernª (Ceratopteris richardii )
Leslie G. Hickok and Thomas R. Warne [full text]
Key Words: fern, sex, plant development, fertilization, genetics, gametophyte, sporophyte.
C-Fern is a simple model plant system based upon the homosporous fern Ceratopteris richardii. C-Fern has a rapid and easily manipulated life cycle that makes it ideal for incorporation into a wide variety of laboratory situations. The small gametophytes afford opportunities for direct observation of developmental events, including spore germination, sexual differentiation, swimming spermatozoids, fertilization and embryo development. The small size and ability to culture large numbers of gametophytes permits easy acquisition of an abundant amount of quantitative data. We will illustrate culture techniques and use of C-Fern in several investigations that highlight development and Mendelian genetics.
[Erratum: In Table 10.A.1 on page 174 of this chapter, K₂HP0₄ is incorrect and should be KH₂PO₄.]

11 -- Pungency Assessment in Onions
Catherine A. Teare Ketter and William M. Randle [full text]
Key Words: plant physiology, pyruvate, Allium, onion flavor chemistry, hydroponics.
The purpose of this investigation is to provide students with experience assaying levels of pyruvate in common onion varieties and onion allies. Additionally, students are asked to modify the protocol to investigate one of several hypotheses relating pyruvate concentration to (1) Allium variety, (2) growth medium sulfur concentration, (3) plant tissue type, or (4) cultivation temperature. An explanation of onion flavor chemistry is included. The laboratory protocol employed is the industrial standard used by commercial onion growers to determine the grade of onion "sweetness". Pyruvate concentration is estimated from a standard curve generated using known pyruvate standards. A hand-held refractometer will provide an estimate of the soluble sugars present in the onion tissue.

Laboratory Exercises in Evolution and Ecology

12 -- Sampling, Density Estimation and Spatial Relations
Maggie Haag and William M. Tonn [full text]
Key Words: dispersion, mark-recapture, quadrat, poisson distribution, performance curve, random distribution, systematic sampling, two-step sampling.
In most field and laboratory experiments on populations, it is often difficult to deal with the total population; instead we sample. For the sample to be truly representative of the entire population, it must be unbiased and it must be adequate in size. This lab deals with these features of sampling while employing techniques used to estimate density and determining the spatial relationships among the organisms in the population being tested. Two common methods of estimating absolute population density are mark-recapture and the quadrate method (or variation thereof). In the first part of the lab we will use the technique of mark-recapture of fathead minnows as a common method of estimating absolute population density. In the second part of the lab zooplankton samples collected previously will be processed to determine the densities of the two major zooplankton groups (cladocerans and copepods) using the quadrat method.

13 -- What Can be Learned about Forest Dynamics from the Age Distribution of Trees?
Sheri L. Gutsell and Edward A. Johnson [full text]
Key Words: age distribution, forest dynamics, Lexis diagram.
Temperate forest trees, because they produce annual rings, allow ecologists to gain an understanding of forest dynamics and succession. In this lab we demonstrate that by coring trees and measuring their basal diameter, the age distribution of live trees, their height growth rates, and their age-diameter relationships can be determined. Cross-dating techniques also are used to show how dead trees can be incorporated into the age distribution. The lab emphasizes the limitations of using only live trees in interpreting forest dynamics and succession.

14 -- Forest Fragmentation in the Rocky Mountains
Rebecca Reed, Jane Beiswenger, and Julia Johnson-Barnard [full text]
Key Words: forest fragmentation, forest management, edge effect, roads, clear-cutting, cumulative effects.
Forest fragmentation is a problem of environmental management faced by land managers, conservationists, and the general public throughout the world. This laboratory exercise introduces students to the issue of forest fragmentation by guiding them through exploration of its origin and threats to the integrity and sustainability of our land resources. Students learn about the effects of clear-cutting, the implications of creating edge habitat in forests, and the contribution of roads to the degree of forest fragmentation.

15 -- Variability and Selection in Natural Populations of Wood Lice
Rudi C. Berkelhamer [full text]
Key words: evolution, isopod, natural selection, predation, selection, variability, wood lice
Students measure a variety of morphological and behavioral traits for locally collected wood lice. They characterize the variability of these traits, make predictions about relationships among them, and hypothesize about the effect of predation on trait variability. They test their predictions by performing and analyzing the outcome of a simulated predation experiment. Each group has a choice of traits to study and analyze leading to a variety of hypotheses and predictions being tested. This simple and inexpensive exercise helps students to appreciate the variability present in natural populations and to understand that variability is crucial to evolution by natural selection.

16 -- Factors Affecting the Spread of Disease in Human Populations
Nancy L. Goodyear [full text]
Key Words: computer simulation, disease, disease carrier, immune system, infection pathogen, re-infection.
The objective of this exercise is to show students how the spread of disease is affected by such factors as the presence or absence of carriers, human vs non-human carriers, population patterns and densities, immune status, and the likelihood of re-infection. Using a computer simulation (a software program entitled "Eppie^TM"), students manipulate a certain disease situation, then collect and analyze data. Students record their findings in a short scientific report. This exercise is very useful for introducing the concept of probability, as well as demonstrating the challenges of disease control.

Laboratory Exercises in Physiology

17 -- The Mammalian Masticatory Apparatus: An Introductory Comparative Exercise
Anthony P. Russell [full text]
Key Words: Mammalia, rabbit, muscles, dissection, mastication, dentition, skull, function.
Exposure to well-circumscribed and tractable exercises is an important part of the training of undergraduates in comparative anatomy. Use of fresh, rather than preserved, material provides a greater appreciation of functional systems. The masticatory apparatus of mammals provides an example of how a musculoskeletal complex can be studied to introduce the basic principles of comparative anatomy and functional morphology. This exercise, based upon fresh rabbit heads (reclaimed from physiology laboratories and thus not requiring sacrifice specifically for this purpose), explores how muscle architecture influences the functioning of the system, and is enhanced by a study of other functional masticatory groups as outlined by W.F. Turnbull (1970, Fieldiana Geol. 18: 149-356). The exercise is designed to be completed in a three hour period.

18 -- Use of Impedance Pneumography in Exercise Physiology
Connie Brewer and Mary Gray [full text]
Key Words: impedance pneumograph, exercise physiology, MacLab data collection/analysis.
This laboratory exercise investigates the changes in the cardiovascular and respiratory systems that occur with exercise. Students initially record heart rate, systolic blood pressure, respiratory rate and depth, and skin temperature for a subject at rest. The subject is then asked to do a timed stepping exercise. The parameters are subsequently measured at the end of exercise and during the recovery period. The impedance pneumograph provides a convenient method of recording the rate and depth of respiration using two trans-chest electrodes. This technique is also applicable to other mammals (i.e. rats). Data acquisition at the workshop will be done with Macintosh computers and MacLabs, although other systems may be utilized.

World Wide Web Information

19 -- Developing Course and Laboratory Homepages for the World Wide Web
Glenn A. Bauer and Glenn MacDougall [full text]
Key Words: Internet, HTML, Netscape Navigator Gold, web page.
The Internet and World Wide Web are increasingly becoming important tools for use in the dissemination of lecture material and laboratory information for teaching biology. This document will introduce the reader to the basic components of a web page, and show them how to create these items themselves through the use of software called Netscape Navigator Gold. This WYSIWYG (what you see is what you get) HTML editor makes web page creation easy, even for the complete novice. Specifically, the readers will learn how to insert images, create links, and format general page appearance to their specifications.

20 -- Enhancing Your Biology Course and Lab WWW Pages with the Latest in HTML, CGI, and JavaScript Features
Glenn MacDougall and Glenn Bauer [full text]
Key Words: HTML, CGI, Java, World Wide Web.
This workshop directed at individuals with basic htm and web page construction experience, demonstrates how to enhance your course webpages. We will demonstrate techniques in a platform independent manner such that you can add tutorials, quizzes, embedded sound, video, image maps and JavaScript to your course homepages. We will discuss the potential and limitations of enhanced webpages and the basic differences between the Internet and your own campus Intranet. We will show how to optimize your pages for delivery in either situation. Templates, and evaluation copies of current shareware applications will be given to participants and will be used during the session to prepare web pages. This session, while not advanced, is not really appropriate for individuals who have little or no background in web page construction.

Mini Workshops