Cell and Molecular Methods and Techniques
BIO 221A --Spring 2011
Instructor:
Office:
Phone:
Office hours:
Email:
Dr. Tom Peavy
SQU 406
278-7276
M/T 3:00-4:30pm (or by appt)
trpeavy@csus.edu
Class Time:
Wednesdays & Fridays 1pm – 3:50pm in SQU 208
Course Content and Objectives:
BIO 221A is a graduate level introduction to research methods in cellular and molecular
biology. Students learn both cell and molecular techniques in the context of hypothesis-driven
research to answer questions relating to a specific gene and cellular system. Experimental
design and commonly used laboratory techniques will be explored.
Lab Manual:
Cell and Molecular Methods and Techniques Lab Manual. CSU, Sacramento. The manual will
be available for download at the course website within SacCT:
https://online.csus.edu/webct/logonDisplay.dowebct .
Course Requirements:
A. Student attendance and participation in each of the laboratory sessions is expected.
Maintaining a personal laboratory notebook is required and is essential for keeping yourself
organized in the lab (will be checked at intervals throughout the semester). Failure to attend
the lab sessions, to participate, or to keep a satisfactory lab notebook will result in the loss
of points at the instructor’s discretion.
B. Grading will be based on the result of two written exams, two laboratory write-ups, the
laboratory notebook, and class participation (includes contribution to PowerPt presentation).
Evaluation
Exams (2, 100 points)
Points
200
Lab Write-ups (2, 75 points each) 150
Laboratory Notebook
25
Class Participation
45
Total Points
420
Bio 221A, Cell and Molecular Methods and Techniques
1
Course Policies:
A. Make-Up Exam Policy:
Make-up exams will be given to a student only if the circumstances surrounding their
absence was beyond their control, as deemed by me. I will require written documentation to
verify the circumstance within one week of the missed exam.
B. Adding/Dropping the Course:
It is important that each student evaluate at the beginning of the semester whether he
or she has the time necessary to devote to the course. The CSUS 2010-11 Class Schedule
details the University’s add/drop policy. In order to drop the course after the Census Date,
students are required to provide documentation to verify their extenuating circumstances.
Forms are available in the Biological Sciences Department office.
C. Classroom Conduct:
Be mindful of potential distractions that you can prevent. Electronic devices should be
off, or, if necessary, set to vibrate. If you need to leave early, please let your instructor and
lab partners know in advance. If you come late to class, do not disrupt the class. Please be
respectful to each other and listen when anyone is asking questions, presenting or when I
am lecturing.
D. Cheating and Plagiarism:
Evidence of cheating or plagiarism, will be reported to the Department Chair and the
Dean of Students, and the student may be dismissed from the course with a grade of “F”.
Other possible lesser actions may be the loss of all points for the assignment/report/exam
and/or reduction of the students final grade. All exams are closed book and notes.
Plagiarism is defined by the University as “the use of distinctive ideas or works belonging to
another person without providing adequate acknowledgement of that person's contribution.”
The university’s policy on plagiarism is provided on the web at
http://www.csus.edu/admbus/umanual/ump14150.htm. The CSUS library web site also has
a number of documents addressing integrity and plagiarism:
http://library.csus.edu/content2.asp?pageID=360.
E. Disabilities:
If you have a disability, please let me know as soon as possible so I can make
accommodations to facilitate your full participation. I will make every attempt to maintain
confidentiality.
Bio 221A, Cell and Molecular Methods and Techniques
2
BIO 221A Spring 2011
Tentative Schedule (subject to change)
Week 1:
Introduction to Cellular & Molecular Methods and Research
Wed (1/26): Introduction to cell & molecular techniques
Friday (1/28): Student discussion on specific techniques and COX
Week 2:
Research Methodology, Cell Culture & Microscopy
Wed (2/2): Discuss and perform cell culture techniques
Friday (2/4): Fluorescence microscopy theory and practice
Week 3:
Immunocytochemistry Experiment
Wed (2/9): Immunocytochemistry (COX-1 & COX-2 antibodies)
Friday (2/11): Fluorescence microscopy imaging
Week 4:
Analysis of Immunocytochemistry Experiment and Protein Isolation
Wed (2/16): Analysis of fluorescence images, troubleshooting, &
design of new round of immunocytochemistry expts
Friday (2/18): Protein isolation from cell culture/ protein quantification
(and freeze cells for mRNA prep)
Week 5:
Revised Immunocytochemistry Experiment
Wed (2/23): Round 2 of Immunocytochemistry (COX-1 & COX-2);
Friday (2/25): Fluorescence microscopy imaging
Week 6:
Protein Electrophoresis and ELISA
Wed (3/2): Introduction of SDS-PAGE/western blotting/ELISA techniques
Friday (3/4): Application of SDS-PAGE/Westerm Blotting/ELISA
Week 7:
RNA Isolation and PCR Design
Wed (3/9): Immunoblotting & Intro to Flow Cytometry
Friday (3/11): Flow Cytometry experiment & RNA isolation from cell culture
Week 8:
Exam 1 and PCR
Wed (3/16): review for Exam 1; RNA quantification
Friday (3/18): PCR design discussion and Reverse transcriptase PCR (RT-PCR);
SPRING BREAK
Wed (3/23) and Fri (3/25)
Week 9: Gel Electrophoresis, DNA Purification, Ligation & Transformation
Wed (3/30): Exam 1
Friday (4/1): Agarose gel electrophoresis/ PCR product purification/
and Ligation
Week 10:
Plasmid Purification, Restriction Digests, Gel Electrophoresis
Wed (4/6): Lab report 1 due; Transformation
Friday (4/8): Plasmid DNA purification and restriction digests
Bio 221A, Cell and Molecular Methods and Techniques
3
Week 11:
Discuss Real Time PCR and Analysis of Sequence Information
Wed (4/13): Agarose gel electrophoresis/ selection of clones for DNA sequencing
Friday (4/15): Discuss Real Time PCR methodology
Week 12:
Real Time PCR
Wed (4/20): Perform Real Time PCR experiment (qPCR)
Friday (4/22): Analyzing DNA sequence information from submitted clones
Week 13:
Design Poster Presentation on COX Experiments
Wed (4/27): Analysis of qPCR data and design new experiments
Friday (4/29): Perform new experiments
Week 14:
Student Presentations
Wed (5/4): Perform new experiments
Friday (5/6): Perform new experiments
Week 15:
Review for Exam 2; EXAM 2
Wed (5/11): Work on Presentations; Review for Exam 2
Friday (5/13): Group Presentations on COX expts; Lab report 2 due
Finals Week
Mon (5/16; 12:45-2:45pm): Exam 2
Bio 221A, Cell and Molecular Methods and Techniques
4