1. 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?

2. 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?

3. 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

4. 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.Basics of PCR 1.Denaturation 2.Annealing 3.Extension http://users.ugent.be/~avierstr/principles/pcr.html

5. 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.

6. 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?

7. 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