1. Jeopardy Final Jeopardy Gene Cloning Plasmids Ligase PCR $100 $100
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Final Jeopardy

2. 1 - $100
When a restriction enzyme cuts parts of DNA, what does it create?
Sticky ends

3. 1 - $200 Where are restriction enzymes found?
They are naturally present in bacteria. Can be taken out to use for gene cloning.

4. 1 - $300 What are restriction enzymes function in the cell?
To guard against viral bacteria – bacterial immune system.

5. 1 - $400 True or False. Restriction enzymes can cut any part of DNA.
False. They bind to certain target sequences in DNA.

6. 1 - $500 How would you come up for the name of this enzyme?
Escherichia coli RI
EcoRI

7. 2 - $100 What is the shape of these small, supercoiled DNA molecules?
Circular

8. 2 - $200 What kind of genes to plasmids have?
Genes that help the cell adapt to unusual environmental conditions.

9. 2 - $300
True or False. They often are passed naturally from one bacterium to another by conjugation or transformation.
Type answer to appear with a mouse-click here

10. 2 - $400
What are two properties that scientists have modified plasmids to have to use in gene cloning?
To contain selectable markers
To contain multiple restriction sites

11. 2 - $500 Name all four advantages for use in gene cloning.
Self replicating
Non essential for cell survival
Can integrate into the bacterial genome
Contain antibiotic resistance genes

12. 3 - $100 What is the function of ligase?
To connect two parts of complementary DNA.

13. 3 - $200
True or False. In gene cloning, you don’t have to add DNA Ligase because it is naturally attached to the Vector DNA.
False. You have to add Ligase to your gene cloning toolkit.

14. 3 - $300
True or False. DNA Ligase only connects fragments that were cut by a certain restriction enzyme.
False. DNA Ligase connects any two complementary bases. Not dependent on the type of restriction enzyme.

15. 3 - $400
About how many different enzyme systems are identified in microbes?
Over 3,000!! WOW

16. 3 - $500 What part of the backbone of DNA does ligase connect?
The sugar phosphate backbone of DNA fragments.

17. 4 - $100
What does PCR stand for?
The Polymerase Chain Reaction.

18. 4 - $200
How many cycles of PCR is needed to make two molecules of your gene of interest?
Three

19. 4 - $300
Around what temperature is needed for the denaturing process, primer annealing, and primer extension.
Denaturing – around 95 degrees C
Primer annealing – around 45 degrees C
Primer extension – around 72 degrees C

20. 4 - $400
What would happen if you lowered the temperature to around 10 degrees C in the primer annealing stage?
It would be too cold and no reactions would happen. Specifically there would be the separation of DNA strands, but the primers could not bind to the template DNA.

21. 4 - $500 Name all five things you need in your PCR Toolkit.
DNA to be copied (template)
DNA Primers
Taq DNA Polymerase
dNTPS (nucleotides)
Buffer solution

22. 5 - $100
True or False. You do not need to perform PCR before using Gel electrophoresis.
False. You need to perform PCR to have the amplified DNA. You need the STR loci.

23. 5 - $200
What is the charge of DNA?
Negative

24. 5 - $300
When you apply an electric field to the gel matrix, does the DNA go towards the positive charge or the negative charge?
Positive, because DNA is negative.

25. 5 - $400
If you find a band of DNA that is extremely thick, what does that mean?
That there is a lot of DNA that is that same mass and same size.

26. 5 - $500
If most of the human genome is identical, why do we find differences between our gel electrophoresis patterns?
Because there is some amount of variation in regions that are not involved in protein coding that we can identify and distinguish between individuals.

27. Final Jeopardy
Describe to me how transgenic animals are generated such that the gene of interest is only expressed in a specific cell type (i.e. mammary cells).
Insert your recombinant DNA into the oocyte of a cell (reproductive cell).