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Engebrecht & Stauffer Title of lesson plan: Speed versus endurance: what can your genes tell you? Misconceptions: Students are confused about the relationship between phenotype and genotype, the proposed lesson plan will distinguish between genotype at the DNA level and phenotype (speed versus endurance). There is widespread belief that genes solely determine phenotype, analysis of the data should reveal that there is a much more complicated relationship between genes and resulting phenotype. State standards: California Life Science/Biology Standard 4. Genes are a set of instructions encoded in the DNA sequence of each organism that specify the sequence of amino acids in proteins characteristic of that organism. As a basis for understanding this concept: 1) students know hoe to apply the genetic coding rules to predict the sequence of amino acids from a sequence of codons in RNA; 2) students know how mutations in the DNA sequence of a gene may or may not affect the expression of the gene or the sequence of amino acids in an encoded protein. California Life Science/Biology Standard 5. The genetic composition of cells can be altered by incorporation of exogenous DNA into the cells. As a basis for understanding this concept: 1) students know the general structures and functions of DNA, RNA, and protein; 2) students know how to apply base-pairing rules to explain precise copying of DNA during semi3) students know how basic DNA technology (restriction digestion by endonucleases, gel electrophoresis, PCR) is used to construct recombinant DNA molecules. Major science concepts: Central dogma (DNARNAprotein) Genetic information passed down from generation to generation. Individual variation and how it may contribute to phenotype Objective of learning outcomes: SWBAT describe important steps in the transcription and translation of genetic code into observable phenotype. SWBAT evaluate the effects of the ACTN3 gene on athletic performance. SWBAT provide pro and con arguments for the use of genetic testing for the ACTN3 gene in humans. List the skills students will use in your teaching plan. 1) Ability to extract DNA 2) A general understanding of gel electrophoresis and PCR 3) Ability to synthesize several basic concepts 4) Use of evidence in making choices 5) Brainstorming and group discussions

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Page 1: List the skills students will use in your teaching plan. 5 ...gena.mspnet.org/media/data/EngebrechtStaufferFR.pdf?media_000000006923.… · We conducted the lesson plan two times,

Engebrecht & Stauffer Title of lesson plan: Speed versus endurance: what can your genes tell you? Misconceptions: Students are confused about the relationship between phenotype and genotype, the proposed lesson plan will distinguish between genotype at the DNA level and phenotype (speed versus endurance). There is widespread belief that genes solely determine phenotype, analysis of the data should reveal that there is a much more complicated relationship between genes and resulting phenotype. State standards: California Life Science/Biology Standard 4. Genes are a set of instructions encoded in the DNA sequence of each organism that specify the sequence of amino acids in proteins characteristic of that organism. As a basis for understanding this concept: 1) students know hoe to apply the genetic coding rules to predict the sequence of amino acids from a sequence of codons in RNA; 2) students know how mutations in the DNA sequence of a gene may or may not affect the expression of the gene or the sequence of amino acids in an encoded protein. California Life Science/Biology Standard 5. The genetic composition of cells can be altered by incorporation of exogenous DNA into the cells. As a basis for understanding this concept: 1) students know the general structures and functions of DNA, RNA, and protein; 2) students know how to apply base-pairing rules to explain precise copying of DNA during semi3) students know how basic DNA technology (restriction digestion by endonucleases, gel electrophoresis, PCR) is used to construct recombinant DNA molecules. Major science concepts: Central dogma (DNA�RNA�protein) Genetic information passed down from generation to generation. Individual variation and how it may contribute to phenotype Objective of learning outcomes: SWBAT describe important steps in the transcription and translation of genetic code into observable phenotype. SWBAT evaluate the effects of the ACTN3 gene on athletic performance. SWBAT provide pro and con arguments for the use of genetic testing for the ACTN3 gene in humans.

List the skills students will use in your teaching plan.

1) Ability to extract DNA

2) A general understanding of gel electrophoresis and PCR

3) Ability to synthesize several basic concepts

4) Use of evidence in making choices

5) Brainstorming and group discussions

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Curriculum materials:

Textbook: Biology: The Dynamics of Life. Glencoe 2005. ISBN 0-07866580-9.

Equipment and materials to extract DNA, perform PCR, restriction endonuclease and gel electrophoresis.

Primary Article: Yang et al., (2003) ACTN3 genotype is associated with human elite athletic performance. Am. J. Hum. Geneti. 73:627-631 (Reference only)

This lesson will take place near the end of the genetics unit from a college prep high school biology course. Students will already have studied the Central Dogma of protein synthesis, Mendelian genetics, and molecular genetics.

Big Idea: Phenotypes are the result of a complex interaction between genes and environment.

For extracting and analyzing students DNA IRB Approval was obtained:

Dear Principal Investigator: This is formal notification that the Institutional Review Board has conducted a review of your research study and relays to you the following action taken by the IRB: IRB Meeting Date: - IRB Protocol Number: 200917599-1 Title of Study: GENA (Geneticists-Educator Network Alliance): Lesson Plan IRB Action: Your study was approved as submitted. Final approval forthcoming in 1-7 working days. This is a courtesy notice from the IRB Administration. Investigators may not respond to IRB concerns through this e-mail communication. Please retain your protocol number for record-keeping purposes both with the IRB Administration and the Sponsored Programs Office/Office of Research.

If you do not wish to receive such e-mail communications from the IRB Administration, please visit http://research.ucdavis.edu/notifications/hsp/noreview_result.cfm?tst=1BD36747-81FB-4855-9F6F557E34E3B4B4

Parent permission form (attached)

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Lesson plan:

Day 1 (30 min)

Pre-Assessment (Stauffer) - attached

Day 2 (90 min)

Introduction Powerpoint (Engebrecht) – attached

Extract DNA from cheek cells (protocol - attached)

Day 3 (90 min)

Analysis of DNA Powerpoint (Engebrecht) – attached

Virtual PCR and virtual gel electrophoresis (http://learn.genetics.utah.edu/content/labs).

Analysis of results and discussion: pro and con use of genetic test for ACTN3 gene.

Day 4 (30 min)

Post test.

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Isolation of DNA from buccal (cheek) cells PCR works effectively to amplify small starting amounts of DNA from any source as long as DNA is not highly degraded and is free of proteins that inhibit Taq polymerase activity. Tissues used as a source of DNA can be fresh (blood, hair root bulbs, sperm, skin tissue, and cheek cells are examples) or relatively ancient (muscle tissue from museum specimens, corpses, and even well-preserved tissue of extinct organisms). For this lab, you will isolate DNA from your own cheek cells obtained by a saline mouth rinse. If you have been eating, go to the drinking fountain and rinse out your mouth before proceeding. 1. Pour 5 ml of 0.9% saline solution into your mouth and swish vigorously for 30 seconds. 2. Expel the solution back into the tube from which it came. 3. Swirl the solution to mix well and transfer 1 ml to a 1.5 ml microfuge tube. 4. Centrifuge for 60 seconds to pellet cells. 5. Pour off the supernatant without disturbing the cell pellet. 6. Add 30 μl of 0.9% saline solution to the pellet and resuspend by pipetting up and down. 7. Add 100 μl of 10% Chelex suspension 8. Vortex to mix cells and Chelex. Attach the special cap lock to your tube and place it in a 99°C heat block for 10 minutes 9. Centrifuge the tube for 2 minutes 10. Transfer 100 μl of supernatant to a clean, labeled tube. In the following part of this exercise you will use 20 μl of your supernatant as template for your PCR reactions.

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Dear Parents/Guardians of _______________________ Your child’s biology class will be participating in a unique genetics lesson involving the University of California, Davis and the Geneticist-Educator Network of Alliances. An important part of the lesson involves extraction of genetic material from cells located inside each student’s cheek. The procedure for obtaining the cells is harmless, painless, and does not result in any bleeding. The genetic material will be used to determine what version of a gene, ACTN3, your child has. This gene has been loosely associated with types of athletic performance. One of the goals of the lesson will be for students, using their own genetic information, to evaluate the possible benefits and controversies surrounding genetic testing. If you have any questions regarding this lesson, please contact me at [email protected] or 916-375-7800. Thank you! Don Stauffer Science Department Chair I grant permission for my child to participate in this genetics lesson. _______________________________ Signature _______________________________ Printed Name _______________________________ Date

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Name: _________________________ Speed Versus Endurance Pre-Test Please write in complete sentences using Standard English. Please do your best. You will be graded on effort – it’s okay if some of your answers are incorrect. 1) Determine this amino acid sequence.

DNA TAC TGT GTT TCA TCA CCC CTC GGA

mRNA

Amino acids

2) Enzymes are what kind of macromolecule? How do geneticists use restriction enzymes? 3) What does polymerase chain reaction do? 4) Is each trait determined by only one gene? Explain. 5) Are there factors other than genes that explain how traits appear? Explain. 6) What do you know about the ACTN3 gene.

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Name: _________________________ Speed Versus Endurance Post-Test Please write in complete sentences using Standard English. Please do your best. You will be graded on effort – it’s okay if some of your answers are incorrect. 1) Determine this amino acid sequence.

DNA TAC TGT GTT TCA TCA CCC CTC GGA

mRNA

Amino acids

2) Enzymes are what kind of macromolecule? How do geneticists use restriction enzymes? 3) What does polymerase chain reaction do? 4) Is each trait determined by only one gene? Explain. 5) Are there factors other than genes that explain how traits appear? Explain. 6) What do you know about the ACTN3 gene.

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7) Explain the arguments for and against the testing of children for the ACTN3 gene.

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Engebrecht & Stauffer – Supplement We conducted the lesson plan two times, the first in the fall and the second in the spring to two different biology classes. Pre and post assessment results are below. In general, we believe that the lesson plan was successful and that the students benefitted from the experience. We learned several things from the first offering:

1) The presentation was too long to keep the students engaged. 2) Extraction of DNA was very chaotic as Engebrecht had an unrealistic expectation of

what the students could accomplish. 3) There was not enough time to adequately discuss the results. 4) The students had not covered enough genetics, especially Mendelian genetics, at the

time of the lesson plan. The second offering went much more smoothly. This was accomplished by the following:

1) The presentation was shortened to keep the students engaged. 2) Stations were set up ahead of time to facilitate the DNA extraction. 3) The presentation was shortened, which provided enough time for discussion of the

results. 4) The students had covered key areas of genetics, including the concepts of homozygous

and heterozygous, in addition to the central dogma, before the lesson was given. Pre and Post assessment results here:

Fall 2009 Spring 2010

Pre-Test Post-Test Pre-Test Post-Test

Total Quizzes Total Quizzes Total Quizzes Total Quizzes

31 32 32 29

Correct Responses

Percent Correct

Correct Responses

Percent Correct

Correct Responses

Percent Correct

Correct Responses

Percent Correct

1) Determine this amino acid sequence

30 96.8% 32 100.0% 32 100.0% 29 100.0%

2) Enzymes are what kind of macromolecule?

24 77.4% 31 96.9% 16 50.0% 18 62.1%

How do geneticists use restriction enzymes?

2 6.5% 12 37.5% 7 21.9% 11 37.9%

3) What does polymerase chain reaction do?

1 3.2% 8 25.0% 1 3.1% 13 44.8%

4) Is each trait determined by only one gene? Explain.

4 12.9% 9 28.1% 5 15.6% 8 27.6%

5) Are there factors other than genes that explain how traits appear? Explain.

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5 16.1% 10 31.3% 3 9.4% 6 20.7%

6) What do you know about the ACTN3 gene?

4 12.9% 24 75.0% 2 6.3% 23 79.3%

7) Explain the arguments for and against the testing of children for the ACTN3 gene.

24 75.0% 25 86.2% Analysis of Pre and Post Tests Virtually all the students could correctly determine the sample amino acid sequences. We analyzed the responses to question two separately (Enzymes are what kind of macromolecule? How do geneticists use restriction enzymes?), as student responses differed between the first and second parts of the question. Students in the Fall biology class were better at describing an enzyme, probably due to differences in what had been presented in their class. However, only a little more than a third of the students in both groups were able to correctly describe the purpose of a restriction enzyme in the Post Test. Question four (Is each trait determined by only one gene?), revealed a couple of entrenched misconceptions. Many students responded that one gene coded for only one trait. Other students confused gene and allele – responding that different traits would be determined depending on the genetic make ups of parents. As might be expected, the Post Test showed that students were much better at explaining the role of the ACTN3 gene, as well as explaining arguments for and against its use in testing children.