Objective 1 - Describe how restriction enzymes are used to manipulate DNA.

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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/genetic
s/biotech/gels/virgel.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?

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Objective 2 - Identify how gel electrophoresis is
used to analyze nucleic acids and proteins and to
distinguish between two alleles of a gene.

• 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?

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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/activitie
  s/blackett/karyotype.html

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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.
• Denaturation
• Annealing
• Extension
• http//users.ugent.be/avierstr/principles/pcr.htm
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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.

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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 victims DNA.
• Sample 2 is the crime scene evidence.
• It is a mixed blood sample of
• the victims DNA and the killers DNA.
• Sample 3 is a sample of the suspects DNA.
• Sample 4 is the control.
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• Questions
• A. Is the suspect guilty of the crime?
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• B. How do you know?
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Objective 9 - Explain how short tandem repeats
(STRs) are analyzed to determine a persons
genotype. Objective 10 - Explain how STRs are
used to identify a persons 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/activitie
  s/blackett2/overview.html