BIO 2265: SPECIAL TOPICS: CELL BIOLOGY LABORATORY SPRING 1999

LABORATORIES:	Lewis, Room 213
	Wednesdays 2:00-5:00

LABORATORY TEXT: none

INSTRUCTOR:	Randall Morrison, Ph.D.
	Lewis, Room 213 A
	Office phone: 410-857-2409
	E-mail: rmorriso@mcdaniel.edu

Office hours: Tuesday, Wednesday and Thursday from 1:00-2:00; and/or whenever else you can find me in 213A. If these times do not work for you feel free to call and make an appointment with me at any time.

DATE	TOPIC
W Jan. 27	Module I: Succinate dehydrogenase background
W Feb. 3	Module I: Fractionation
W Feb. 10	Module I: Enzyme assays
W Feb. 17	Module II: Zebrafish fibroblast culture background
W Feb. 24	Group report I due (on Module I)
	Module II: culturing
W Mar. 3	Module III: FITC capping background
W. Mar. 10	Individual assignment I due (on Module II)
	Module III: FITC experiment
W. Mar. 17	Spring Break&emdash;No labs this week
W. Mar. 24	Module IV: Alu fingerprinting background
W. Mar. 31	Group report II due (on Module III)
	Module IV: Alu fingerprinting experiment
W. Apr. 7	Module V: Isolation and quantitation of embryo RNA
W. Apr. 14	Individual assignment II due (on Module IV)
	Module V: Preparation of the blot and probe labeling
W. Apr. 21	Module V: Hybridization and detection
W. Apr. 28	Practical exam
	Module VI : Degenerate oligonuculeotide PCR
W. May 5	Group report III due (on Module V)
	Module VI : Degenerate oligonucleotide PCR

1. Attendance: Attendance of laboratories is mandatory. Given the structure of the course described below it will not be possible to "make-up". Any potential conflicts need to be explored both the instructor and your group very early in the term.

2. Module topics: Module I will address enzyme function and activity by investigating succinate dehydrogenase. Differential centrifugation will be used to isolate the enzyme and then enzyme activity will be assessed spectrophotometrically. Module II will involve tissue culture techniques. We will attempt to establish primary cell cultures of fibroblasts and pigment cells from zebrafish fins. Module III will address protein movement within membranes. We will use fluorescently labeled antibodies crosslink membrane proteins into capping structures. Module IV will introduce molecular techniques including PCR. We will amplify DNA from each individual in the class for a neutral polymorphism found in humans. In Module V will attempt a northern blot to examine the expression of a gene during the embryogenesis of zebrafish. In Module VI we will use PCR to attempt to amplify novel pigment related genes in zebrafish using primers designed from other vertebrates.

3. Laboratory assigments and groups: During the first week of lab we will be establishing laboratory groups that will work together for the entire semester. You will be conducting all of the experiments as a group and three of the five lab assignments are lab reports your group will prepare. It is important that your lab group function well. Over the course of the semester if group dynamics become an issue you should come and talk to me (preferably before the report is due!). Your group will be designing your own experiments to some extent, preparing to conduct them, and then conducting the experiment. In order for groups to be effective every member will have to contribute to the process.

Scheduling and time required in the laboratory will be rather flexible and often group dependent. Some experiments may end sooner or take longer than you anticipate. You will also need to periodically schedule times to get together outside of lab to work on the group reports.

Three lab reports are to be entirely group generated. Each group will produce one report for which I will assign a group grade. The expectation is that everyone will contribute to the process: preparation, experimentation and report. I will be asking each group member for a paragraph describing their contribution and other members contributions to the work. 75% of the grade will be the group grade and 25% will be based on both my perception and the group perception of contribution to the project.

The other three lab assignments will be individual efforts. The experiments will be conducted in groups but each group member will be expected to generate an independent project. These will not be entire lab reports, but a project that relates to preparing reports. Examples may include preparing a lab protocol as opposed to a Materials and Methods section, or preparation of written results and a discussion given a common Introduction, Materials and Methods and figures provided to you. I will explain the nature of each of these assignments as they approach.

Lab reports and assignments will be due at the beginning of the indicated lab session (at 2:00 sharp). Attached to this syllabus is an explanation of lab report format and the forms that will be used to score lab reports.

4. Practical exam: There will be one practical exam that will be designed to test your facility with all the equipment and techniques that we will be using in the lab. You are responsible for knowing how to use all of the equipment and techniques used regardless of whether you conducted a particular part of an experiment. This exam is scheduled as shown near the end of the term.

5. Grading: The breakdown of the grade is specified below:

Group report I	17%
Group report II	17%
Group report III	17%
Individual assignment I	10%
Individual assignment II	10%
Practical exam	29%
	100%