M, W 8:00 - 10:50 Science B Rm 122

Course Description: The course objective is for you to gain hands-on experience with fundamental techniques used in contemporary Cell Biology research. We will carry out an ongoing, coordinated project involving diverse experimental methodologies which are applicable to many of the different fields within Cell Biology. Our focus will be the cloning and expression of a eukaryotic protein using a baculovirus expression system. Techniques you will use include construction of recombinant bacmids, eukaryotic cell culture, protein purification and analysis, polyacrylamide gel electrophoresis, Western Blot analysis, immunolocalization and PCR. 6 (+) hours laboratory/wk.

Textbook: None. Protocols, background, and discussion material will be provided in the form of handouts.

Grades: A total of 400 points can be earned in the laboratory section of BIO410, which is half of the total points for the class. The point breakdown is as follows:

Points Component

200 4 (50 point) quizzes

150 3 (50 point) research papers

25 1 lab report

25 participation/attendance, notebook

Attendance: Is required and monitored -- points will be deducted for absences not pre-arranged with the instructor.

Lab Notebooks: A bound (not spiral) lab notebook is required for this course. It must be brought to every class, and it must be kept up to date. This lab notebook should have an up-to-date table of contents, and it should be clear enough in organization that your instructor can pick up the notebook and understandwhat you have done. These notebooks will be spot-checked during the semester and collected at the end of the semester. These lab notebooks will be used for the preparation of 3 detailed research papers.

Research Papers: Research Papers are to be written in journal format, as prescribed by the journal Molecular Biology of the Cell. These reports should be prepared following the instructions to authors from this journal. These research papers will be focused on the results of experiments that you have carried out in the class. In addition to the these three research papers, you are expected to prepare a laboratory report on the last few experiments of the semester. A reprint from this journal is posted outside my office door (Science B 227) for you to use as a reference. You can also download PDF versions of articles from a sample issue of the journal which can be accessed at the journal's homepage.

A link to the instructions to authors can be found here:

Instructions for downloading your gel images and importing them into text files can be found here:

http://www.humboldt.edu/~msw13/410_html/image1_html/howto1.html

Images for your research papers can be found here:

http://www.humboldt.edu/~msw13/410_html/image1_html/imagindex1.html

links to images (including SEM pics) for the third paper can be found here

Quizzes: The last three quizzes will address your understanding of the procedures that we have covered in the lab section of the class. The first quiz will assess your ability to make up simple solutions in the lab, and to solve problems using serial dilution theory. On the website for the class you will find a variety of sample problems on these topics

The first quiz will be on Wednesday, 1-26-00. This quiz will be focused on calculations for making solutions, and on serial dilutions. The problems will very similar to the sample problems found at the following pages:

Serial dilution theory and sample serial dilution problems with answers

sample solution problems with answers

This is a tentative schedule. The work performed in this lab is sometimes difficult to place on an exact schedule, but we will attempt to stay as close as possible to the above timing.