CHEM 419:010 Biochemical Methods
Tues, Thurs
Dr. Katherine R. Miller email: krmiller@salisbury.edu
332 Henson Science Hall phone: x36481
Objectives: Through laboratory work and discussion, the student will experience many common techniques involved in biochemical and molecular biological research. This is a workshop style course which means most of the work will be hands on. It is expected that the students will ask questions and initiate discussions relating to the material as they proceed.
Textbook: Scheppler, Judith A., Patricia E. Cassin,
and Rosa M. Bambier.
2000. Biotechnology
Explorations. ASM
Press,
Office hours:
Mondays and
Wednesdays
Attendance Policy: Many of these experiments build off of one another. In some instances, you will be working in pairs and in others, individually. If you miss a day,you will either need to arrange to make that day up as soon as possible outside of class time or ask your lab partner to work alone and share his/her data with you. There are no make up days for this class.
Academic Integrity: Please read the university’s academic integrity policy in the student handbook on pages 51-52. This handbook can be found at: http://www.salisbury.edu/Students/handbook/. If you are having difficulty with an assignment, please talk to me rather than giving into the temptation to cheat. If you are caught cheating in any way, it may result in failure of the course.
Accommodations for
students with disabilities: If you
need accommodations, please talk to me within the first few weeks so that we
can make appropriate arrangements.
Grading: Grading is on a ten point scale:
100%-90% A
89%-80% B
79%-70% C
69%-60% D
59% and below F
Final averages will be rounded to the nearest whole number.
Each project report will consist of:
a concise objective statement 10 pts
an analysis of the procedure 30 pts
results:
quality 20 pts
analysis 20 pts
example applications in
the literature 20 pts
Grades will be calculated as follows:
WebCT reading quizzes: 10 %
3 projects (20 % each) 60 %
midterm 15 %
final 15 %
Tentative syllabus:
1/27: Introduction to Bioinformatics
Assignment of project 1: design of PCR primers
2/1 Exercise 25: Gene analysis
2/3 Exercise 26: Sequence Analysis and Sequence Alignment
2/8 Exercise 27: Primer design
2/10 Free day to work on project 1
2/15 Project 1 report due, discussion of project 1, Introduction to DNA preparation and Analysis
2/17 Preparation of solutions and materials for next few exercises.
2/22 Exercise 7: Restriction analysis of l-DNA
2/24 Exercise 9: Transformation of bacteria with pGreen plasmid
3/1 Finish Exercise 9, prep for exercise 8
3/3 Exercise 8: plasmid miniprep
3/8 Exercise 22: Isolation of genomic DNA - day 1
3/10 finish exercise 22
3/15 Exercise 23: Comparison of DNAs from the Five Kingdoms - day 1
3/17 finish exercise 23
3/21 - 3/26 spring break
3/29 Midterm exam
3/31 Project 2: Isolation and characterization of DNA from soil
Isolation of DNA from soil using kit, set up PCR reactions
4/5 Clean up PCR reactions, digest with enzymes
4/7 Analyze PCR reactions and digests, clone and transform PCR products
4/12 finish project, analysis of results.
4/14 Project 3: Purification and Characterization of Green Fluorescent Protein
Introduction to protein purification and characterization
inoculate cultures with transformed bacteria
computer analysis of protein structure
4/19 Project 2 report due, purification of GFP
4/21 purification of GFP
4/26 purification of GFP
4/28 exercise 17 - determination of protein concentration
5/3 exercise 18 - SDS-PAGE of GFP
5/5 enzymatic digestion and analysis of GFP structure
5/10 finish project
5/12 Project 3 report due
5/16 Final exam