Chabot College

Course Outline for Biotechnology 40, Page 1

Fall 2010

Chabot CollegeFall 2010

Course Outline for Biotechnology 40

Biotechnology Laboratory Skills I

Catalog Description:

40 – Biotechnology Laboratory Skills I4 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: Mathematics54(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):

  1. Scientific method, data and summary statistics
  2. Measurements and metric system
  3. Solutions and dilutions
  4. Introduction to spectrophotometry
  5. Basic chemistry; pH and buffers
  6. Membrane transport
  7. Organic chemistry:macromolecules
  8. Chromatography
  9. Enzymes kinetics
  10. Cell biology
  11. Cell reproduction
  12. Introduction to microbiology
  13. Mendelian genetics
  14. Molecular inheritance
  15. Recombinant DNA technology
  16. Introduction to PCR
  17. Genomics
  18. Proteomics
  19. Introduction to immunology

Course Content (Laboratory):

  1. Laboratory safety
  2. Maintaining a laboratory notebook
  3. International system of measurement; unit conversions; scientific notation
  4. Laboratory glassware; measuring temperature, mass, volume, and length
  5. Constructing tables and graphs
  6. Preparing solutions
  7. Preparing stock solutions and buffers
  8. Serial dilutions
  9. Using micropipetters
  10. pH measurement
  11. Spectrophotometry
  12. Bradford assay for protein quantification
  13. Microscopy
  14. Microbial cultures
  15. Aseptic culture of E. coli
  16. Bacterial streaking and staining techniques
  17. Overnight liquid culture of bacteria
  18. DNA extraction and characterization
  19. Extraction of genomic DNA from E. coli
  20. Agarose gel electrophoresis of DNA
  21. UV spectrophotometric quantification of DNA
  22. Recombinant DNA methodology
  23. Restriction endonuclease digestion of λ DNA
  24. Determine size of restriction gragments
  25. Restriction mapping
  26. Plasmid transformation with pGLO with E. coli host
  27. Polymerase chain reaction
  28. DNA extraction from human cheek cells
  29. PCR of Alu PV 92 and agarose gel electrophoresis analysis
  30. PCR of mtDNA D-Loop and agarose gel electrophoresis analysis
  31. DNA genomics
  32. mtDNA analysis with databases
  33. Protein extraction and purification
  34. Thin layer chromatography (TLC)
  35. Column exchange chromatography
  36. Protein purification
  37. SDS-PAGE
  38. Western blotting

Methods of Presentation:

  1. Cooperative laboratory activities
  2. Instructor demonstrations
  3. Media presentations
  4. Lectures and discussions

Assignments and Methods of Evaluating Student Progress:

  1. Typical Assignments
  2. 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.

  1. Writing

1)Maintaining laboratory notebook

2)Worksheets, e.g., solving problems and completing study guide reviews

3)Laboratory reports

  1. 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

  1. Methods of Evaluating Student Progress
  2. Examinations, including a final examination, combination of short answer, fill-ins, multiple choice, matching, and essay
  3. Professional quality laboratory notebook
  4. Quizzes, homework, class participation

Textbooks (typical):

Essential Biology, Campbell, N., Reece, J., and Simon, E., Benjamin Cummings, 3rd edition, 2007

Basic Laboratory Methods for Biotechnology, Seidman, Lisa A. and Moore, Cynthia J., Prentice Hall, 2nd edition, 2009

Special Student Materials:

  1. White laboratory coat
  2. Safety goggles
  3. Disposable latex gloves
  4. Bound laboratory notebook
  5. Sharpies for marking glassware and Petri dishes
  6. Black ink pens for notebook

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