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Home > Contents
of Proceedings > Volume 17
Tested Studies for Laboratory Teaching
Volume 17
Purdue University, June 6-10, 1995
Editor: Jon C. Glase (Cornell University)
Host: Susan J. Karcher
1. Expression Vectors Used in Project-Oriented Teaching
Laboratories by Susan J. Karcher, David J. Asai, and Chris
J. Staiger, Purdue University [abstract]
[full text]
2. Heads or Tails? Patterns of Segmental Regeneration
in a Freshwater Oligochaete by Charles D. Drewes, Iowa
State University [abstract]
[full text]
3. Chemical Properties of Amino Acids and Identification
of Unknown Amino Acids by Sam Donovan, Carl Stiefbold,
and Karen Sprague, University of Oregon [abstract]
[full text]
4. Reporter Genes and Transgenic Plants to Study Response
to Environmental Signals by Stanton B. Gelvin and Susan
J. Karcher, Purdue University [abstract]
[full text]
5. Immunofluorescence of Cytoskeletal Proteins by
John C. Mordacq and Roberta W. Ellington, Northwestern University
[abstract]
[full text]
6. Water and Solute Movement Through Red Blood Cell Membranes
by Richard S. Manalis, Indiana University-Purdue University
[abstract]
[full text]
7. Reproductive Isolation in Angiosperms by Alexander
F. Motten, Duke University [abstract]
[full text]
8. Use of Macintosh Computers and Maclab Data Acquisition
Units in a Human Cardiovascular Lab by Philip J. Stephens,
Villanova University [abstract]
[full text]
9. Using Microcomputer-Based Physiology Experiments in
Investigative Labs in Introductory Biology by Robert J.
Kosinski and Jean L . Dickey, Clemson University [abstract]
[full text]
10. Techniques of Anesthesia and Catheter Placement in
the Rat (Rattus rattus) by Mary H. Gray, Purdue
University [abstract]
[full text]
11. Identification of Microorganisms Encountered in the
Upper Respiratory Tract by Iris L. Sun, Purdue University
[abstract]
[full text]
12. Life-History Schedules in Daphnia magna: An
Ecological Activity for Multiple Laboratory Sessions by
Robert E. Sorensen, Purdue University [abstract]
[full text]
13. Investigations of Relationships among Wetland Organisms
Using Pickle Jar Aquaria by Ann B. Burgess and Anne
F. Dehring, University of Wisconsin-Madison
[abstract] [full
text] [app. b]
14. The Estimation of Species Richness in Pennsylvanian
Coal Swamp Communities by Gary E. Dolph, Indiana University
Kokomo [abstract]
[full text]
Appendix A: Abstracts of Mini Workshops [titles]
[full text]
Abstracts (Vol. 17)
Laboratory Exercises in Cell and Molecular Biology, Genetics,
and Development
1 -- Expression Vectors Used in Project-Oriented
Teaching Laboratories
Susan J. Karcher, David J. Asai, and Chris J. Staiger
[full text]
Key Words: expression vector, E. coli, protein isolation,
SDS-polyacrylamide gel electrophoresis
One lab of a multi-week project taught in an upper level cell
and molecular biology laboratory is given. Isolation of sufficient
quantities of rare proteins from biological tissues can be
difficult. These native proteins are often required for raising
antibodies, and for studying enzymatic or regulatory function.
Expression vectors are used to obtain large quantities of
proteins. The cDNA encoding a protein is cloned into an expression
vector. The fusion protein is over-expressed in E. coli
, and large quantities of the fusion protein are obtained
by affinity chromatography. Participants isolate proteins
from E. coli, and separate proteins by SDS-polyacrylamide
gel electrophoresis.
2 -- Heads or Tails? Patterns of Segmental
Regeneration in a Freshwater Oligochaete
Charles D. Drewes [full
text]
Key Words: regeneration, pattern formation, morphallaxis,
development, Lumbriculus variegatus, annelid
The freshwater oligochaete, Lumbriculus variegatus,
is introduced as a new model for student investigations of
animal regeneration and pattern formation. Advantages of this
species include: (1) relatively long body (= 4-5 cm), (2)
ease of culture and handling in the lab, (3) robust and relatively
rapid regeneration of amputated head and/or tail segments
(e.g., dozens of new tail segments form within 2-3 weeks),
(4) high survival rates in excised body fragments of all sizes,
(5) consistent and quantifiable patterns of segment regeneration
in newly formed heads and tails, (6) easily observed internal
organ systems in intact worms or worm fragments, and (7) morphallactic
reorganization of segments within the original body fragment.
3 -- Chemical Properties of Amino Acids
and Identification of Unknown Amino Acids
Sam Donovan, Carl Stiefbold, and Karen Sprague [full
text]
Key Words: amino acid structure, ionization, pH, titration
curves, ion exchange chromatography, CPK models.
These laboratory activities are designed to engage students
in a set of abstract concepts which form the basis for understanding
protein structure and function. Through model building and
problem solving activities students develop and apply their
understanding of many important biochemical concepts including:
amino acid structure; physical properties such as size and
hydrophobicity; and the relationships between pH, pKa and
the net charge of an amino acid. The summative task involves
designing an experimental protocol to identify unknown amino
acids based on an understanding of how they will respond to
ion exchange chromatography, titration, gel filtration, and
electrophoretic analysis. While this is not a traditional
"wet" lab, the presentation of the material is structured
so that students must apply their knowledge of the concepts
in order to determine the identity of their unknowns.
4 -- Reporter Genes and Transgenic Plants
to Study Response to Environmental Signals
Stanton B. Gelvin and Susan J. Karcher [full
text]
Key Words: reporter genes, transgenic plants, Agrobacterium
tumefaciens, GUS assay, b-galactosidase assay.
In these laboratories, students work with gusA reporter
genes in transgenic plants and lacZ reporter genes
in bacteria. A reporter gene encodes an enzyme that is easily
assayed. Students observe simple color changes to study the
induction of promoters of plant genes in response to environmental
(cold stress and gravitropism) and developmental stimuli.
Students also study the response of bacterial gene promoters
to chemical signals from plants. The response of these promoters
is studied by assaying reporter gene activity. These reporter
gene assays are suitable for the undergraduate teaching laboratory
because they are easy to perform, reliable, safe, and visible.
5 -- Immunofluorescence of Cytoskeletal
Proteins
John C. Mordacq and Roberta W. Ellington [full
text]
Key Words: immunofluorescence, cytoskeleton, antibodies,
tubulin, actin.
This exercise presents the technique of immunofluorescence
and how it is used to detect and localize specific proteins
or molecules within a cell, specifically, the cytoskeletal
proteins, tubulin and actin. This procedure is made possible
by using antibody molecules that selectively recognize and
bind specific antigens. Mouse fibroblast cells, grown in culture,
will be fixed in paraformaldehyde; incubated with primary
antibodies directed against cytoskeletal proteins; and then
incubated with secondary antibodies that are conjugated to
one of two different fluorescent molecules (FITC or rhodamine).
The preparation will be viewed using a Leitz Biomed fluorescent
microscope.
Laboratory Exercises in Physiology
6 -- Water and Solute Movement Through
Red Blood Cell Membranes
Richard S. Manalis [full
text]
Key Words: RBCs, tonicity, osmosis, membrane permeability,
solute transport, transient turbidity measurements, hemolysis
time, computerized data acquisition.
Students are presented with several action statements which
challenge them to use their critical thinking skills to design
specific protocols for the study of the osmotic behavior of
RBCs under various experimental conditions. Experiments performed
with nonpermeable solutes pertain to osmosis and tonicity,
whereas those performed with permeable solutes pertain to
relative permeability measurements and, thus, to solute transport
across the RBC membrane. Membrane permeability for a given
solute is measured indirectly as the hemolysis time for that
solute. The hemolysis time is obtained with a modern data
acquisition system consisting of a Spectronic 20D interfaced
with a MacLab/Macintosh computer.
7 -- Reproductive Isolation in Angiosperms
Alexander F. Motten [full
text]
Key Words: angiosperms, isolating mechanisms, pollen tubes,
pollination, reproductive isolation, Silene.
During this exercise students investigate pre-zygotic isolating
mechanisms in three species of Silene (catchfly, campion).
In the experimental portion, students use flowers of S.
alba to compare pollen tube growth in styles following
pollination from S. alba, S. dioica, and S.
noctiflora. These results are then combined with information
on habitat, flowering time, and pollination syndromes. S.
alba is a particularly convenient experimental subject
because it is dioecious, the five styles are completely separate,
and the flowers are large enough to allow easy manipulation
and impressive pollen tube growth.
8 -- The Use of Macintosh Computers and
Maclab Data Acquisition Units in a Human Cardiovascular Lab.
Philip J. Stephens [full
text]
Key Words: breathing, lung volumes, MacLab data acquisition
unit, gravity, heart cardiovascular function.
Simple spirometry permits a student volunteer to measure tidal
volume and inspiratory and expiratory reserve volumes. Measurements
are made at different body positions to examine the effects
of gravity on the different lung volumes. Heart function is
monitored by recording either the electrocardiogram or finger
pulse. This open-ended portion of the laboratory allows students
to explore cardiovascular function during normal breathing,
slow breathing, after exercise, and the effects of high carbon
dioxide and low oxygen levels on cardiovascular physiology.
9 -- Using Microcomputer-Based Physiology
Experiments in Investigative Labs in Introductory Biology
Robert J. Kosinski and Jean L . Dickey [full
text]
Key Words: investigative, microcomputer-based labs, physiology,
interfacing, transducers, introductory.
In investigative labs, students design their own experiments.
Using experiments based on physiological interfacing, we introduced
an investigative format into the second semester of an introductory
biology course for non-majors. We produced our own transducers,
interfacing software, and lab manual. Working in groups, students
devised experiments, presented proposals, performed their
experiments, and then reported on the results. Students and
instructors liked the new lab format. There were some indications
from a pre- and posttest on science process skills that the
investigative students developed their process skills more
than the students still taking the traditional course. Lecture
scores of the investigative and traditional students were
the same.
10 -- Techniques of Anesthesia and Catheter
Placement in the Rat (Rattus rattus)
Mary H. Gray D.V.M. [full
text]
Key Words: anesthesia, vascular cannulization, rat, surgery,
euthanasia.
Rats are extremely useful animals for laboratory instruction
in mammalian physiology. They are readily obtained, relatively
inexpensive, and easy to maintain. Students are able to learn
rodent restraint, anesthetic protocol, and basic surgical
techniques with minimal difficulty. The ability to isolate
the vagus nerve and to cannulate vessels is utilized in most
physiology laboratories. For example, vagal stimulation is
used to demonstrate bradycardia and heart block while the
ability to deliver fluids and diuretics IV is essential to
renal function investigations.
11 -- Identification of Microorganisms
Encountered in the Upper Respiratory Tract
Iris L. Sun [full
text]
Key Words: Gram Staining, Corynebacterium, Haemophilus,
Neisseria, Staphylococcus, Streptococcus.
The upper respiratory tract (URT) forms a network of interconnected
passageways harboring a large and primarily normal flora which
in some occasions may carry pathogens and constitutes a reservoir
of infection. The mouth especially contains a complex mixture
of bacteria, all present in very large numbers: 50 to 100
billion bacteria/ml of saliva and 25 to 45 billion/mg dental
plaque. In contrast to most other diseases, the same organisms
of the normal flora are found in both healthy and sick individuals.
The development of a pathogenic process is not necessarily
the result of the introduction of new bacteria, but of a rupture
of equilibrium in favor of larger microbial populations as
a result of poor oral hygiene. Similarly the nose flora and
the oropharynx flora contain organisms associated with the
mouth. In addition, bacteria which preferentially attach to
the epithelium are potential pathogens. Below the larynx,
the respiratory tract is devoid of microorganisms and is thus
normally sterile. It is therefore important to learn the characteristics
of microorganisms encountered in URT and how to identify them.
The objective of this workshop is to let participants be able
to recognize and identify the major types of microorganisms
(such as Streptococci, Staphylococci, Haemophilus,
Corynebacteria and Neisseria) associated with
the throat, mouth, and nose.
Laboratory Exercises in Evolution, Ecology, and Behavior
12 -- Life-History Schedules in Daphnia
magna-An Ecological Activity for Multiple Laboratory Sessions
Robert E. Sorensen [full
text]
Key Words: life-history theory, fecundity, survivorship,
population growth, trade-offs, net reproductive rate.
This lab exercise introduces students to the ecological principles
influencing population growth by engaging them in a three-week
study of population growth of Daphnia magna populations
housed at three densities. The students work cooperatively
in small groups to collect fecundity and mortality data daily
from their populations. In the end, the groups pool their
data and construct life-tables for the three treatment groups
to address hypotheses about the effects of density on survival,
reproduction, population growth and life-history trade-offs.
The culmination of this project is a scientific paper written
by each student presenting their findings.
13 -- Investigations of Relationships
among Wetland Organisms Using Pickle Jar Aquaria
Ann B. Burgess and Anne F. Dehring
[full text] [appendix
b]
Key Words: wetland ecology, food webs, predator-prey relationships,
model system, open-ended project.
This project involves a field trip to a local marsh where
students (1) become familiar with a wetland ecosystem, and
(2) collect materials they use back in the lab to assemble
aquaria in one gallon glass pickle jars. The pickle jar aquaria
are the basis for a semester-long open-ended project in which
students investigate relationships among wetland organisms
using observations and simple tests.
14 -- The Estimation of Species Richness
in Pennsylvanian Coal Swamp Communities
Gary E. Dolph [full
text]
Key Words: coal swamps, paleobotany, paleoecology, palynology,
Pennsylvanian, species richness, Westphalian.
The use of a nonparametric statistic, the jackknife, to estimate
species richness in Pennsylvanian coal swamp floras is discussed
here for the first time. Answers to a number of significant
paleoecological questions can be sought using this technique.
Did species richness vary from place to place in a coal swamp?
Did species richness increase through geologic time as suggested
by a number of authors? Were coal swamp communities open or
closed communities? This technique is suitable for use in
an honors laboratory or (with suitable modifications) in any
freshmen biology laboratory in which basic ecological principles
are investigated.
Mini Workshops
[full text]
- Getting the Message Out: Technology Equipment for
Chemistry and Biology Classrooms (Diane W. Burnett)
- Introductory Molecular and Cell Biology Courses-Open
General Discussion (Anne Cordon)
- Simulating a Pond System Over 24 Hours (Jim Eckblad)
- Animals as Factors Shaping Plant Communities (Roy
Hurst)
- Using Commercially Available Human Cells in a Chromosome
Lab (Rosamond Potter)
- A Hands-On Simulation of Disease Transmission
(Jean Dickey)
- The Use of Digital Imaging in General Botany Laboratories
at The University of Wisconsin-Madison (Mike Clayton)
- Micro-Techniques of Cell Harvesting in Mice (Frances
F. Makowski)
- Simple Software to Demonstrate Changes in Allele Frequency
(Susan M. Schenk)
- A Play for Genetics Lab (Roberta B. Williams)
- The Role of Biologists in Science Education of Elementary
and Middle School Teachers (Ned Lyke)
- Chemotaxis in Physarum, a Plasmodial Slime Mold (a
Simple Experiment to Teach Chi2 Analysis) (Donna M.
Bozzone)
- Don't Say Math in a Biology Class (Laurie Iten)
- Education and the Internet (Edward Andrews)
- Introducing Students to Scientific Literature
(Nora Ann Bennett)
- Using an Investigative Approach to Teach the Concepts
of Fermentation (Arthur L. Buikema Jr., Suzanne H. Braunschweig,
and Donna Harpold)
- The Use of Research, Teaching, or Personal Collections
to Teach an Inquiry Based Introductory Biology Lab (Suzanne
H. Braunschweig and S. Llyn Sharp)
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