MODULE 2
Objective 2.2 Lesson A
RFLP
Capstone of Basic Biotechnology Skills
Course
Advanced
Biotechnology
Rationale
In molecular biology, restriction fragment length polymorphism, or RFLP, refers to
small differences in otherwise highly similar DNA sequences.
Unit
In RFLP analysis, the DNA sample is broken into pieces (digested) by restriction
enzymes, and the resulting restriction fragments are separated according to their
lengths by gel electrophoresis. RFLP analysis was the first DNA profiling technique
inexpensive enough to see widespread application. In addition to genetic
fingerprinting, RFLP was an important tool in genome mapping, localization of genes
for genetic disorders, determination of risk for disease, and paternity testing.
DNA Analysis
Essential
Question How
do scientists
sequence DNA,
and what are
some
implications of
this technology?
An example of student work analyzing RFLPs is below:
TEKS
130.364 1A-K,
2F-J, 3A-C, 7F,
9A, 9C, 9F, 10C,
12B
TAKS
Science 1A, 2A,
2B, 2C, 2D
Prior Student
DNA structure,
micropipette,
electrophoresis,
restriction
enzymes
Estimated Time
2 hours
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Objectives
Students will:
 Describe how RFLPs are generated.
 Explain RFLP analysis applications.
 Interpret RFLP analysis results using different scenarios.
 Demonstrate proficiency in Biotechnology Skill Standards.
Key Points
I. See Power Point for Lesson Two Objective 2.2
Engage
 Student complete the Blackett Family DNA Activity-TFLP Analysis
Activity
1. Wet Lab: There are many simulated RFLP kits sold through your
district approved Science vendor. Examples include:
• Crime scene analysis
• DNA fingerprinting
• Cancer gene detection
• Cholesterol Diagnostics
• Sickle cell detection
2. Chose several kits that you think your students would be
interested in and have groups work on different applications. If
your total enrollment i n the course is low and you will not use
all of these samples, they can be stored in the refrigerator for up
to 2 years for future use.
3. This activity provides a capstone f or students to work
independently in t he lab with little help from you to showcase
their lab techniques, research skills and technology skills. This
activity is appropriate for a fall exam project.
4. Assign a different case from the above lab kits to each lab
group.
5. Generate a list of “Key Words/Concepts” associated with each
case from the teachers’ manual for each case for students to
guide them in their research. Students should use these key
words to research how their topic is detected using RFLP. A
“case” is provided in each kit manual to give to students. This
also provides a nice opportunity to work with your school
librarians to find scientific journals available to students.
Copyright © Texas Education Agency 2012. All rights reserved.
6. Give students a protocol from each lab manual and have
students carry out the experiment. They should not need any
help from you.
• Tip: Photos of gel can be taken with any smartphone and
emailed for editing.
• Tip: Use PAINT to edit gel picture (label, change colors,
make bands darker).
7. Use the “Biotechnology Skill Standards Practical Rubric” to
grade students’ performance throughout the lab. Students
should c all on you when they are ready to demonstrate mastery
of a skill. Information on Skill Standards can be found at
http://www.tssb.org/.
8. Allow students to generate their own poster rubrics. See
“Student Sample Work” for ideas.
9. Print out posters and allow students to peer grade using studentgenerated rubric.
• Printing is set for poster printing on templates.
• Kinko’s will also print poster size if you email them your
file.
• If you do not have access to color printing, upload poster
to a file share application and have students present and
peer grade.
10. Hold a poster session for other students in your school. Great
way to promote your biotechnology course for future enrollees!
Materials
• Worksheet: “Blackett Family DNA Activity-RFLP Analysis”
• Handout: “Case Study Instruction Sheet”
• Handout: “Case Study Grading”
• Handout: “Biotechnology Standards Practical”
Assessment
• Correctly answer questions in Blackett Family DNA Activity
• Case Study Grading Handout
• Biotechnology Standards Practical
• Student generated Poster Rubric
Copyright © Texas Education Agency 2012. All rights reserved.
Accommodations for Learning Differences
 Visit the Special Populations section of the CTE Career and
Technical Education Website: http://cte.unt.edu/special-pops.
National and State Education Standards
Texas College and Career Readiness Standards
I. Nature of Science: Scientific Ways of Learning and Thinking
C1, C2, C3, D1, D2, D3, E1, E2
II. Foundation Skills: Scientific Applications of Mathematics
A2, A5, A7
III. Foundation Skills: Scientific Applications of Communication
A1, B1, B2, B3, D1
Copyright © Texas Education Agency 2012. All rights reserved.
Blackett Family DNA Activity-RFLP Analysis
Name:_______________
Go to http ://www.biology.arizona.edu/humanbio/activities/blackett/introduction.htm l.
Click on the link, “RFLP Analysis,” read the information, then click on “detailed
description of VNTR.” Next, answer the questions below.
Questions:
1. According to the website, VNTR stands for "variable number of tandem
repeats." What does that mean?
2. What does Restriction Fragment Length Polymorphism Analysis do?
3. From the Restriction Fragment Length Polymorphism Analysis page, click
“NEXT,” scroll to the bottom and complete the Karyotyping Activity.
a. What notation would you use to characterize Patient A’s karyotype?
What diagnosis would you give patient A?
b. What notation would you use to characterize Patient B’s karyotype?
What diagnosis would you give patient B?
c. What notation would you use to characterize Patient C’s karyotype?
What diagnosis would you give patient C?
4. Go back to the original website after completing the Karyotyping Activity
(see address above), then click on “RFLP Analysis.” How do scientists
determine how RFLP markers are named?
5. Click “NEXT,” then read the information. Why is "Length Polymorphism” of
RFLP important? What do the bands represent?
6. Click “NEXT.” Why don’t married couples share any band s? Click
“NEXT.”
Copyright © Texas Education Agency 2012. All rights reserved.
7. How many bands do most people have?
8. Describe how the bands represent inheritance in children. How does this
relate to the number of bands most people have?
9. Click “NEXT.” Why do analysts use a control? What are ladders used for?
10. Answer the following questions about the DNA profiles. Use the hints as
needed!
a. Are the grandparents maternal or paternal?
b. The autorad contains 8 alleles for the siblings tested.
Which of the alleles and how many are shared between each of the siblings?
c. Are any of the unknowns related to the family? If so, which ones?
d. Are any of the other unknowns tested related to each other? If so, which
ones?
e. Are there any 1-locus matches between non Blackett family members?
11. Briefly describe how RFLPs can be used in a paternity case
Copyright © Texas Education Agency 2012. All rights reserved.
RFLP Case Study Instructions
Who are your team members? ___________________________________________
What case have you been assigned? ______________________________________
ALL materials generated must be saved on a flash drive or to a web file storage location.
1. Using the “Key Words/Concepts” list given to you by your instructor, research the
science behind your case study. Generate a list of questions your team has for
your case. Be sure to include how electrophoresis will be used in detection. List
your questions below:
2. Schedule a meeting with your instructor to discuss your questions and receive
approval to move forward. Teacher signature of approval :___________________
3. Collect resources to answer your team questions. Resources must include at least
one of the following (list them below):
a. An online source: _______________________________________
b. A journal article:_________________________________________
c. A book: _______________________________________________
d. A periodical source: _____________________________________
4. Schedule a meeting with your instructor to discuss your questions and receive
approval to move forward. Teacher signature of approval: __________________
5.
6.
7.
8.
Obtain your sample and protocol from your instructor.
Carry out lab protocol.
Photograph Results (phones work fine)
Create a Scientific Poster using technology
a. Visit http://www.ncsu.edu/project/posters/NewSite/index.html for advice on
creating scientific posters.
b. Many universities and public domains can free poster templates that you
can download and enter your information in. Search the Internet for a free
template.
9. Create Poster Rubric.
10. Download a temple, enter your information, save it, and email to your instructor or
save it to a shared drive. Ask you instructor how he/she would like for you to share
your work.
Copyright © Texas Education Agency 2012. All rights reserved.
Case Study Grading
Name:_____________________
I. List of Questions Generated
Teacher Signature: _____________________________
Comments:
II. Resources Collected
Teacher Signature: _____________________________
Comments:
III. Poster Rubric Submitted
Teacher Signature: _____________________________
Comments:
IV. Biotechnology Skill Standards Practical Total Points: _ _
V. Poster Total Points:
Comments:
Copyright © Texas Education Agency 2012. All rights reserved.
Biotechnology Skill Standards Practical
•
•
•
Name :_______________________
Below are selected Biotechnology Skill Standards found at www.tssb.org.
Throughout the Case Study team members should provide evidence of proficient for each standard. Team
members choose what evidence to provide.
Standards will be scored on a scale of 1-10 with 10 being excellent and 1 being unacceptable.
Skill Standard
Evidence
A3. Operate equipment
B2. Perform assays and experiments
B3. Troubleshoot experiments and
equipment
Copyright © Texas Education Agency 2012. All rights reserved.
Team Score
(self
assessment)
Teacher
score
B4. Perform data analysis
B5. Communicate results
C5. Coordinate with work team
C7. Handle and dispose of
hazardous materials
D2. Create documents
Copyright © Texas Education Agency 2012. All rights reserved.