Course title

Pre-requisite

Course description

This course is designed to provide students with a hands-on laboratory experience in the field of biotechnology. Students spend more than half of their time in the laboratory preparing; conducting and analyzing experiments. All lab preparations and clean-up are conducted by the students.

Course Outcomes:
1. Use current biotechnology techniques to investigate/solve real world scenarios.
2. Analyze and communicate the findings of scientific investigations related to biology and
biotechnology.
3. Apply knowledge of biotechnology techniques to make ethical decisions which impact
themselves; society and the environment.
4. Utilize technology and inquiry to increase understanding of biological concepts and issues.
5. Explore various careers and skills associated with biotechnology in the workforce.

Labs:

Formulating Solutions
Microbiological Investigations
Plating bacteria
Streaking Bacteria
Transforming colonies
Electrophoresis
DNA extraction & isolation
Restriction Enzyme digestion
Isolation of DNA/ Fingerprinting
Protein extraction/Proteomics
Microarrays
Polymerase Chain Reaction amplification of DNA

Using standard Scientific Method outlined by the following questions; describe one typical laboratory assignment associated with this course.

HUNTINGTON?S DISORDER LAB:

State the question or concept investigated during this laboratory assignment.
This lab will test a family of four; mother; father; daughter and son; for the presence of the Huntington?s disease gene.
What is the incidence of the Huntington?s Disease gene in a given family consisting of 4 members (father; mother; daughter; son)?

Formulate a hypothesis for the question.
Students formulate alternative hypotheses based on information provided about the family in question and general background information in genetics.
(Based on symptomatic information the mother has Huntington?s disease gene and one or both children will also have the Huntington?s disease gene.)

Formulate a prediction based on the hypothesis using ?if/then? statements.
If the individual possesses the Huntington?s disease gene and the DNA is subjected to gel electrophoresis; then a band will form closer to the well then the individuals lacking the gene.

Describe the experiment you performed to prove or disprove your hypothesis. List all essential materials. Describe each step you performed in the experiment.

Materials:
DNA samples: mom; dad; son; daughter
Agarose
TAE buffer solution
Micropipets and tips
Electrophoresis apparatus

Procedures:
1. Obtain all of your materials and remove the comb and masking tape from the gel tray.
2. Place the gel tray into the electrophoresis chamber; making sure the loading wells are on the black/negative side.
3. Add enough TAE buffer solution to completely cover the gel.
4. Load 14 ?l of each DNA sample into different wells in the following order: Marker; mom; dad; son; daughter.
5. Place the lid onto your gel box (red to red; black to black); being careful not to bump the gel too much. Plug the power source in and set it to 100 V. Turn it on and run the gel for approximately 15 minutes.
6. While the gel is running; make some predictions: Based on what you know about how electrophoresis separates DNA fragments; predict how a marker containing the following sizes of DNA nucleotides will move through the gel. Marker: 30 nucleotides; 60nt; 90nt; and 120nt. Indicate your prediction on the gel below in the lane labeled marker:

Marker Healthy Huntington?s

7. Now; using what you know about the size of the gene for Huntington?s disease; predict where a DNA sample from a healthy person will be after you run the gel and do the same for a person with the Huntington?s gene in the gel above.
8. After your 15 minutes are up; draw a picture of your observations from your gel in your lab book.
9. Use the marker; which tells you the approximate number of nucleotides; to determine the size of each person?s alleles. Record this in your lab book under your picture.
10. Based on the number of nucleotides in the region of DNA; determine the number of CAG codon repeats each person has. Record this in your lab book.
11. The information you analyzed in #10 should help you determine if anyone has the Huntington?s gene and should be used in writing your discussion.

Describe the results of your experiment or study. Use graphs and charts where appropriate.

Explain your data or results. Give an analysis of your experiment.
Huntington?s disease is caused by expansion of polymorphic trinucleotide repeat (CAG)n located in the coding region of the gene for Huntington?s. The range of repeat numbers is 9 to 37 in normal individuals and 37 to 100 in HD patients.

Both the mom and the son have a region of DNA that is larger than 120 nt which is equivalent to 40 amino acids; glycine in the Huntington protein. More tahtn 37 glycines results in possible onset of the disease; with an increased possiblility the greater the number of repeats. The mom and the son have much greater than 40 repeats and therefore would be expected to exhibit Huntington?s The dad and the daughter have 30 repeats or less in their alleles and therefore would not be expected to show symptoms.

Write a conclusion for your study. Was your hypothesis supported or refuted?
Students will follow the usual format for a conclusion indicating if their hypothesis was supported or refuted; indicating who does and doesn?t have the Huntington?s gene with supporting evidence from the experimental results. In addition; students will indicate the genotypes of the various family members with evidence for these claims.

Application Questions:
1. Since Huntington?s disease is a dominant trait; how many of these mutated alleles does a person need in order to get it?

2. If a person heterozygous for Huntington?s disease marries a homozygous normal person; what are the chances of having a child that will develop Huntington?s disease?

3. People that have this trait would likely choose not to have children due to the risk of passing it onto their kids. However; many people pass this trait onto their offspring before realizing they even have it. Why does that usually happen?

4. Does the Huntington?s gene have more or less nucleotides in it than the non-Huntington?s gene?

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Approved

Approved competency code

• LBIO
• Biology

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Online / Virtual