Objective 1 - Describe how restriction enzymes
are used to manipulate DNA.
Run the virtual gel electrophoresis at this web site
http://www.vivo.colostate.edu/hbooks/genetics/biotech/gels/vi
rgel.html
Use the linear genomic clone and load each well with all the
available digests.
Run the animation until you achieve clear and distinct
separation.
Using your gel, explain where each enzyme cut and how that
allowed the bands to separate on the gel electrophoresis.
Question : What do restriction enzymes allow scientists
to do? Hint : Would we be able to do things like DNA
profiling without them? Why or why not?
Objective 2 - Identify how gel electrophoresis is used
to analyze nucleic acids and proteins and to
distinguish between two alleles of a gene.
1. Compare the movement of DNA and RNA
through a gel matrix with the movement of
proteins through a gel matrix.
a. What causes separation of DNA and RNA
fragments?
b. What causes the separation of proteins?
c. Which of these techniques separates two
alleles?
Objective 3 - Define RFLP
analysis.
Objective 4 - Determine how
RFLP analysis can be used to
diagnose genetic disorders.
Complete the Blackett Family
DNA RFLP Activity
http://www.biology.arizona.edu/human_bio/activities/blackett/karyotype.html
Objective 5 - Explain the three main steps of
each cycle of PCR amplification and what
reactions occur at each temperature.
Go to Basics of PCR, read the description of PCR, watch the
animation, and describe what happens at each of the
following steps. Include the relative temperatures and the
reasons for these particular temperatures.
1.
Denaturation
2.
Annealing
3.
Extension
http://users.ugent.be/~avierstr/principles/pcr.html
Objective 7 - Explain the
significance of PCR testing.
1. Why is PCR a more accurate and
practical method than RFLP?
2. What regions of the genome would
you target for profiling? (Think of all
possible answers for this)
3. How many loci are necessary to
establish a legal “DNA fingerprint”
that is used by the FBI and is
admissible in court.
Objective 8 - Analyze and
evaluate real-time PCR results.
These are the results of a DNA test from a
crime scene.
Sample 1 is a sample of the victim’s DNA.
Sample 2 is the crime scene evidence.
It is a mixed blood sample of
the victim’s DNA and the killer’s DNA.
Sample 3 is a sample of the suspect’s DNA.
Sample 4 is the control.
Questions:
A. Is the suspect guilty of the crime?
B. How do you know?
Objective 9 - Explain how short tandem repeats (STR’s) are
analyzed to determine a person’s genotype.
Objective 10 - Explain how STR’s are used to identify a person’s
identity by analyzing a DNA sample.
Read the overview of STR and apply it in
the activities at the hyperlink below
Blackett Family STR analysis – paternity
testing and “missing person” DNA Activity
http://www.biology.arizona.edu/human_bio/activities/blackett2/overview.html