1. Biotechnology

2. Bell Work 1.You want to determine if a patient with leukemia has a mutation in a certain gene. What type of technology should you use and why? 2.If you want to understand how a certain mutation is altering the function of a cell, what type of technology should you use and why?

3. 1. Polymerase Chain Reaction (PCR) - What it is:  Amplifies a segment of DNA (makes lots of copies)  Can amplify a few nucleotides or 1000s of nucleotides - 1 gene or many genes  Imitates natural DNA replication - uses the same enzyme (DNA polymerase)

4.  The DNA is mixed with DNA polymerase, nucleotides and primers.  The mix is alternately heated and cooled in a thermocycler.  Primers: small pieces of nucleotides that bind to the part of the DNA to be amplified How it works:

5. PCR: Primers

6. PCR: Amplification of DNA

7.  Sequence it to check for mutations  Cut it with restriction enzymes to put into a bacterial plasmid  Use it to identify someone (DNA fingerprinting/profiling) How you use the amplified DNA:

8. 2. DNA Profiling or Fingerprinting: Used to distinguish between individuals of the same species using only their DNA Each of us has a unique sequence of DNA Chances of two people having exactly the same DNA profile is 30,000,000,000 to 1 (except for identical twins).

9. Variable Number of Tandem Repeats (VNTR): Repeats of short DNA sequences o The number of repeats varies between individuals. o Each person has many of these repeats scattered in DNA

10. Restriction Fragment Length Polymorphism (RFLP) DNA is isolated from two sources so that they can be compared. The DNA is cut using different restriction enzymes. Different size fragments are produced depending on each persons VNTR.

11. Gel Electrophoresis Used to detect the presence of DNA in a sample. DNA fragments are placed in a gel. Electricity is applied to gel. Negatively charged DNA moves to the positive pole. DNA fragments are separated based on size.

13. The smaller DNA fragments travel farther in the gel. To see the DNA an picture is made using x-rays or radioactive material. The reveals a pattern of bands showing the location of each of the different sized DNA fragments. This picture of DNA bands is called a DNA fingerprint.

14. DNA fingerprinting can be used to determine paternity cases.

15. DNA fingerprinting can be used to identify a suspect in a criminal case.

16. DNA fingerprinting can be used to determine relationships between similar species.

17. Capillary Electrophoresis

18. Capillary Electrophoresis: DNA Sequencing

19. Capillary Electrophoresis:DNA Fingerprinting

20. Bell Work You want to solve a crime using DNA you obtained from the crime scene. You’ve isolated the DNA from the samples obtained. 1.What is the first thing you need to do to the DNA? 2.Why does the DNA move through the agarose gel? 3.What determines how quickly the DNA fragments move through the agarose gel?

21. DNA fingerprint analysis: 1.Plug in the light box and put the plastic bag with your gel on top – turn on the light. 2.Use a ruler to measure the distance each band migrated in each lane and complete the table (#3). 3.Draw a picture of your gel using measurements (#4). 4.Answer the questions on the back of the sheet. 5.DO NOT TAKE GELS OUT OF BAGS. 6.DO NOT DAMAGE GELS – IF YOU DO YOUR TEAM WILL LOSE POINTS ON THE LAB.

22. Ethical, Legal and Social Implications of Genomics: 1.In your group read through and discuss each of the 10 vignettes presented. Use the questions at the end of each to guide your discussion. 2.As a group, select 3 vignettes and write (on a separate sheet of paper): a)Describe 1 positive implication suggested by the vignette. b)Describe 1 negative implication suggested by the vignette.

23. Bell Work Put your backpacks and other bags UNDER your table. You will need a pencil EACH person will complete an analysis sheet!