Chabot College
Fall 2010
Course Outline for Biotechnology 40
Biotechnology Laboratory Skills I
Catalog Description:
40 – Biotechnology Laboratory Skills I
4 units
Introduces students who are interested in biotechnology, biological sciences, and current industry
workers to laboratory research methods and concepts in biotechnology. Laboratory skills include use of
measuring equipment, volume and mass measurements, proper use of micropipettors, pH meters,
spectrophotometers, and microscopes. Additional laboratory skills include sterile techniques, solution and
media preparation, solution dilution, aseptic technique, culture of microbial colonies, agarose and
polyacrylamide electrophoresis, chromatography, DNA extraction, DNA restriction digest, PCR, and
bacterial transformation. Strongly recommended: Mathematics 54 (completed with a grade of “C” or
higher) or appropriate skill level as demonstrated by the mathematics placement test, and eligibility for
English 1A. 2 hours lecture, 5 hours laboratory.
[Typical contact hours: lecture 35, laboratory 87.5]
Prerequisite Skills:
None
Expected Outcomes for Students:
Upon completion of this course, the student should be able to:
1. demonstrate appropriate behaviors, teamwork, and proper safety procedures to work in a
laboratory environment, including maintaining a professional quality laboratory notebook;
2. demonstrate an understanding of the scientific method, experimental design, data collection,
basic statistics, basic laboratory skills, and procedures including the preparation of reagents
and other materials;
3. demonstrate the ability to make solutions, reagents, buffers;
4. demonstrate basic concepts and applications of chemistry and biochemistry appropriate for a
biotechnology laboratory, with the goal of preparing students to work with basic and
sophisticated instrumentation in a biotechnology laboratory, e.g., spectrophotometers,
electrophoresis apparatus, pH meters, and chromatographic systems;
5. demonstrate extraction of DNA from E. coli, its characterization and quantification using
agarose gel electrophoresis, and UV spectrophotometry;
6. demonstrate expertise with the use of the classical tool-kit of molecular biology such as
restriction enzymes, vectors, plasmids, host cells, and expertise of the polymerase chain
reaction (PCR) to clone several loci from the human genome;
7. demonstrate the tools of SDS-PAGE and Western Blotting to characterize purified proteins;
8. demonstrate the proper procedures for the aseptic culturing of microorganisms, their
preparation for microscopy (e.g., Gram staining), and their use as vectors in recombinant
DNA work.
Course Content (Lecture):
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Scientific method, data and summary statistics
Measurements and metric system
Solutions and dilutions
Introduction to spectrophotometry
Basic chemistry; pH and buffers
Membrane transport
Organic chemistry: macromolecules
Chromatography
Enzymes kinetics
Cell biology
Cell reproduction
Introduction to microbiology
Chabot College
Course Outline for Biotechnology 40, Page 2
Fall 2010
13.
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Mendelian genetics
Molecular inheritance
Recombinant DNA technology
Introduction to PCR
Genomics
Proteomics
Introduction to immunology
Course Content (Laboratory):
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Laboratory safety
Maintaining a laboratory notebook
International system of measurement; unit conversions; scientific notation
Laboratory glassware; measuring temperature, mass, volume, and length
Constructing tables and graphs
Preparing solutions
a. Preparing stock solutions and buffers
b. Serial dilutions
Using micropipetters
pH measurement
Spectrophotometry
a. Bradford assay for protein quantification
Microscopy
Microbial cultures
a. Aseptic culture of E. coli
b. Bacterial streaking and staining techniques
c. Overnight liquid culture of bacteria
DNA extraction and characterization
a. Extraction of genomic DNA from E. coli
b. Agarose gel electrophoresis of DNA
c. UV spectrophotometric quantification of DNA
Recombinant DNA methodology
a. Restriction endonuclease digestion of λ DNA
b. Determine size of restriction gragments
c. Restriction mapping
d. Plasmid transformation with pGLO with E. coli host
Polymerase chain reaction
a. DNA extraction from human cheek cells
b. PCR of Alu PV 92 and agarose gel electrophoresis analysis
c. PCR of mtDNA D-Loop and agarose gel electrophoresis analysis
DNA genomics
a. mtDNA analysis with databases
Protein extraction and purification
a. Thin layer chromatography (TLC)
b. Column exchange chromatography
c. Protein purification
d. SDS-PAGE
e. Western blotting
Methods of Presentation:
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4.
Cooperative laboratory activities
Instructor demonstrations
Media presentations
Lectures and discussions
Chabot College
Course Outline for Biotechnology 40, Page 3
Fall 2010
Assignments and Methods of Evaluating Student Progress:
1. Typical Assignments
a. Reading
1) Textbook
Read chapter 1 and summarize the biotechnology applications used in
medical/veterinary and agricultural/food related applications.
2) Articles
Summarize the genesis of the human genome project to the discovery of DNA
discussed in this article. Also provide specific examples on how this revelation
enabled researchers to understand that human genes are responsible for specific
traits.
b. Writing
1) Maintaining laboratory notebook
2) Worksheets, e.g., solving problems and completing study guide reviews
3) Laboratory reports
c. Activities
1) Preparing graphs, tables, and calculations in analysis of laboratory results
2) Measuring mass and volumes to prepare stock solutions and serial dilutions
3) Properly utilizing and maintaining laboratory equipment, e.g., microscopes,
spectrophotometer, micropipettors, pH meters
4) Aseptic culturing and staining of microorganisms
5) Accurate laboratory observations
2. Methods of Evaluating Student Progress
a. Examinations, including a final examination, combination of short answer, fill-ins, multiple
choice, matching, and essay
b. Professional quality laboratory notebook
c. Quizzes, homework, class participation
Textbooks (typical):
Essential Biology, Campbell, N., Reece, J., and Simon, E., Benjamin Cummings, 3 rd edition, 2007
Basic Laboratory Methods for Biotechnology, Seidman, Lisa A. and Moore, Cynthia J., Prentice
Hall, 2nd edition, 2009
Special Student Materials:
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White laboratory coat
Safety goggles
Disposable latex gloves
Bound laboratory notebook
Sharpies for marking glassware and Petri dishes
Black ink pens for notebook
pw G:\Chabot\Biotech
10/2009