Course title
Biotechnology 1-2Pre-requisite
"B" in Biology or Honors BiologyCourse description
Biotechnology 1-2 is offered in the following Tempe Union HS District High Schools: Corona del Sol; Desert Vista; Marcos de Niza; McClintock; Mount Pointe; Tempe HS
Biotechnology 1-2 (BIO107) Curriculum
Quarter 1
Focus Questions Concepts Activities & Labs
What do we need to know to get started? (1 wk) Ice Breakers
Safety Black Boxes/Mystery Cylinders
Room Map
Safety Scenarios
MSDS & Safety Rules
What is biotech? (2 days) Biotechnology definitions Timeline puzzle
How is science done? (1 wk) Inquiry investigation Denim processing lab (open-ended for SOP technique; reference Daugherty text)
SOP?s
What are the basic tools of biotechnology?
(4 wks) ? Chemistry Basics & Biomolecules
? Basic technology: micropipets; vortexers; centrifuges; balances ; UV-Vis Spectrophotometer Micropipet lab (UC Davis protocol)
Measuring Volumes & Mass (Biotechnology: Science for a New Milennium labs 3a-d)
Making Solutions (labs 3e-h)
Milk enzyme lab (lab 1c)
Carbohydrate lab (lab 2e)
Apple Juice Production (lab 5a)
Sugar/Starch Assay (lab 6b)
What are the living tools of biotechnology? (2 wks) ? Cells & Microbes
Basic technology
? sterile technique
? mass/volume calculations
? working with agar
? hot plates & microwave Using the microscope (lab 2c; 2d/Musheno protocol)
Sterile technique & Pouring plates
Culturing E. coli and Saccharomyces
Spectrophotometer for cell density counts
Quarter 2
What do biotechnologists need to know about genetics? (6 wks)
DNA Basics:
? Structure
? Replication
? Gene Expression
? Gene Regulation & Epigenetics
? Recombinant DNA
? Protein Structure
? Cell Signaling
Genetic engineering (prok/euk) DNA Extractions: open inquiry
Protein lab (lab 2f)
Got Protein? (BioRad kit for Bradford dye and Spectrophotometer analysis)
Egg Protein Investigation (application of Bradford procedure; Musheno)
Bacterial Transformation (BioRad pGLO kit)
Plasmid Math
How can biotechnologists examine DNA? (2 weeks) Basic technology
? Horizontal electrophoresis Pouring gels
Electrophoresis of dyes (Musheno)
Restriction digest of DNA and Forensic DNA Fingerprinting (BioRad kit)
Quarter 3
Finish previous unit if necessary
Working with Proteins
(3-4 wks) ? Isolation of molecules using biochemistry
? Antibodies in Biotechnology
Basic technology:
? Affinity chromatography
? SDS-PAGE
? Ouchterlony
? Western Blot ? Protein separation of GFP using Chromatography (BioRad Hydrophobic Interaction Chromatography columns)
? Itchy Pup Ouchterlony lab (mush/Dougherty)
? ELISA for HIV fluid exchange activity (BioRad kit for ELISA)
Biotechnology and Society
(4 wks) ? Basic technology: PCR
? Genetically Modified Organisms in Agriculture
? Intellectual Property & Biotech
? Bioethics Principles ? Harvest of Fear/Future of Food documentaries
? GMO Lab: using PCR and electrophoresis to identify GM foods (BioRad GMO Investigator)
? GM Foods Case Studies: Bt toxin
Quarter 4
How can we judge whether biotechnology should be used? Bioethics case studies Genetic counseling dilemmas; Terri Schiavo
Ocatmom and IVF
How do biotechnologists manage all the information produced? (4 days) Bioinformatics databases Bioinformatics with Gene Gateway: tutorial and investigation using NCBI databases
How can we reduce the costs of biotech at DV?
(1 wk) Plasmid recovery project ? Plasmid mini-preps from Honors Bio transformation labs
? Bacterial Transformation to check results
How can biotechnology help with environmental problems?
(3-4 wks) Bioremediation
Basic technology:
? PCR for identification
? DNA Sequencing (proposed addition for 2011; with primers supplied by David Rhoads at University of Arizona; extension of Mesa High Biotech Academy grant-funded research program)
? NCBI databases ? DV Bioremediators lab: open ended inquiry to search for local soil microbes that can metabolize motor oils (Musheno)
? Identification of soil bacteria (Xan and Dr. Rhoads at UA)
? Analysis of sequence data (NCBI databases)
How can biotechnology be used to improve human health? (2 wks) Reproductive Technology
Pharmacogenomics
Gene Therapy ? Octomom
? Pharmacogenomics case studies
? Gene therapy research
What are the risks of biotechnology? Biological Warfare
Cautionary Tales ? Student research projects
LAB
How Can Electrophoresis be used?
Introduction
In 1950 Oliver Smithes invented the technique of gel
electrophoresis. It quickly proved to be one of the most
powerful tools in the rapidly developing field of biotechnology.
Objectives
1. To investigate the properties of dyes.
2. To understand how electrophoresis can be used to analyze molecules.
3. To learn how to load and run an electrophoresis gel.
Procedure
Day One:
1. Obtain your electrophoresis chamber; and place a casting tray; two dams and a comb appropriately. For this lab; place the comb at the middle position. Why are we using a comb; and why are we placing it in the middle for this investigation?
Be sure the comb is seated securely in the slots provided; and check to see that the number of teeth/wells will be adequate for the number of samples we will load.
2. Prepare 100 ml of 1% agarose using agarose powder and 1x TAE buffer. Write your calculations here for preparing this solution:
3. Heat this mixture on the hot/stirrer plate; using the magnetic stirrer or swirling gently every 30 seconds. Watch carefully for boiling; agarose is prone to superheating followed by violent ?boilover? when it is disturbed. Be sure you have HotHands nearby before you begin!
4. Bring the solution to a gentle boil; and check to be sure all agarose has dissolved in the buffer. The solution should be crystal clear with no ?floaties? when you look through it.
5. Remove agarose from heat and place it on the counter; cool to approximately 50oC (check with forearm test). If you are in a hurry; you can run cool tapwater over the outside surface of the flask while swirling the molten agarose.
6. As soon as the solution reaches 50oC ; pour the agarose into the electrophoresis casting tray. The tray should be about æ full.
7. Allow the gel to solidify (agarose will become cloudy as it cools); then carefully remove the comb (pull straight up) and place the gel in labeled plastic wrap or a ziplock bag to prevent dehydration. Use a glove and the technique demonstrated by your teacher!
8. Carefully rinse out the gel casting tray components (do NOT use a brush on the chamber); and set them on paper toweling to dry.
Day Two:
9. Place your prepared gel in the electrophoresis apparatus; using a casting tray. Fill the electrophoresis chamber with 1x TAE buffer so the gel is submerged by approximately 1mm.
10. Place 10-20?l of each dye sample into separate wells. Be sure everyone in your group loads at least one sample! Record the placement of each sample in your lab notebook.
11. Attach the electrodes to the apparatus; and run the gel for 15 minutes at 100 volts. How can you tell that the connection is good and there is an electric current in the tray?
12. Detach the power supply; remove the gel; and place it in a weigh boat.
13. Examine your gel. You may want to use a lightbox to see the bands more clearly. Take note of which samples moved; and in which direction. Measure and record the distance traveled; from the leading edge of the well to the leading edge of the sample band.
14. Clean up your equipment and gel as directed by your instructor.
15. Create a bar graph to display the distance each band moved from neutral position; be sure your graph will display the direction of movement! Discuss these results with your group and write your conclusions for the lab; be sure to include your answers to the analysis questions below.
Questions to Consider
? How do the electrophoresis chamber and the agarose gel assist a scientist in gathering data about macromolecules?
? Why is the buffer solution used to fill the chamber; rather than plain water?
? Why do lab technicians try to avoid air bubbles when pouring an electrophoresis gel?
? Did all of the dyes run out in the same direction? Why or why not?
? Did all of the dyes run out to the same distance? Why or why not?
? What can we conclude about each dye sample; based on your data? Think of as many ways to explain these results as you can. Be sure your evidence will support these claims!
School Country
United StatesSchool state
ArizonaSchool city
TempeSchool Address
500 W Guadalupe RoadSchool zip code
85283Date submitted
Approved
YesApproved competency code
- CTE
- Career and technical education
- LADV
- Advanced science
- LBIO
- Biology