Chabot College
Fall 2010
Course Outline for Biotechnology 50
Biotechnology Laboratory Skills II
Catalog Description:
50 – Biotechnology Laboratory Skills II
2 units
Introduces students who are interested in biotechnology, biological sciences, and current industry
workers to the advanced laboratory research methods and concepts in biotechnology. Laboratory skills
include mastering the tools used in biotechnology such as isolation and quantification of DNA,
amplifications with PCR, media preparation and dilution, aseptic technique, and cell culture. Prerequisite:
Biotechnology 40 (completed with a grade of “C” or higher). 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. 1 hour lecture, 3 hours laboratory.
[Typical contact hours: lecture 17.5, laboratory 52.5]
Prerequisite Skills:
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.
Expected Outcomes for Students:
Upon completion of this course, the student should be able to:
1. demonstrate understanding of SOPs;
2. demonstrate ability to prepare media, gels, buffer solutions and reagents of specific
concentrations by using the appropriate stock solutions;
3. isolate and purify DNA from biological systems and plasmid vectors from bacterial for cloning;
4. demonstrate the proper use of spectrophotometry to assess the purity and quantity of DNA
that have been isolated, and to determine population density of bacterial cultures;
5. prepare competent cells for transformations with recombinant vectors and calculate
transformation efficiency;
6. analyze and interpret results from electrophoretic gels;
7. construct standard curve graphs relating known DNA fragment size in the estimation of size
of unknown DNA fragments;
8. construct restriction maps from data obtained from electrophoresis of restriction digests;
9. properly prepare and use selective media (with antibiotics as the selective base) to eliminate
nontransformed specimens (level one screening);
10. use the products of PCR as a source of DNA for cloning specific genes from other organisms;
11. design PCR primers on the basis of phylogenetic and nucleotide sequence information
gathered from electronic databases;
Chabot College
Course Outline for Biotechnology 50, page 2
Fall 2010
12. use Internet databases to gather information on the nucleotide sequence of specified genes,
the amino acid sequence of specified proteins, and other molecular information;
13. use alignment software to align DNA sequences downloaded from Internet databases.
Course Content (Lecture):
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Review of laboratory skills
Review of measurements
Bacterial culturing techniques
Solution preparation
DNA restriction digestion and agarose gel electrophoresis of DNA
Effects of DNA methylation on restriction enzyme digestion
Purification, identification, and quantification of plasmid DNA
E. coli DNA
Genomic library
Restriction mapping of circular and linear DNA
PCR primer design using BLAST
Extraction and analysis of RNA
Course Content (Laboratory):
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Micropipetting and sterile techniques
Bacterial culture techniques
DNA restriction and electrophoresis
DNA methylation and restriction
Transformation
Purification and identification of plasmid
pBR322 plasmid restriction mapping
Construction of λ genomic library
Purification of λ clones
Primer design
Optimization of PCR for 12S rRNA primers
Extraction and analysis of RNA
Cell culture
Methods of Presentation:
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Cooperative laboratory activities
Instructor demonstrations
Media presentations
Lectures and discussions
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
Chabot College
Course Outline for Biotechnology 50, page 3
Fall 2010
3) Laboratory reports
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
c.
Textbooks (typical):
Laboratory DNA Science, Bloom, M., Freyer, G., and Micklos, D., Benjamin/Cummings, 1st
edition, 1996*
Basic Laboratory Methods for Biotechnology, Seidman, Lisa A. and Moore, Cynthia J., Prentice
Hall, 2nd edition, 2009
*
This is the most current edition of the book. It has basic and easy to follow laboratory
practices which are currently used in the biotechnology industry.
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