Jeopardy Gene Cloning Plasmids Ligase PCR DNA Fingerprint $100 $100 $100 $100 $100 $200 $200 $200 $200 $200 $300 $300 $300 $300 $300 $400 $400 $400 $400 $400 $500 $500 $500 $500 $500 Final Jeopardy
When a restriction enzyme cuts parts of DNA, what does it create? Sticky ends 1 - $100
1 - $200 • Where are restriction enzymes found? • They are naturally present in bacteria. Can be taken out to use for gene cloning.
1 - $300 • What are restriction enzymes function in the cell? • To guard against viral bacteria – bacterial immune system.
1 - $400 • True or False. Restriction enzymes can cut any part of DNA. • False. They bind to certain target sequences in DNA.
1 - $500 • How would you come up for the name of this enzyme? Escherichia coli RI • EcoRI
2 - $100 • What is the shape of these small, supercoiled DNA molecules? • Circular
2 - $200 • What kind of genes to plasmids have? • Genes that help the cell adapt to unusual environmental conditions.
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
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
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
3 - $100 • What is the function of ligase? • To connect two parts of complementary DNA.
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.
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.
3 - $400 • About how many different enzyme systems are identified in microbes? • Over 3,000!! WOW
3 - $500 • What part of the backbone of DNA does ligase connect? • The sugar phosphate backbone of DNA fragments.
4 - $100 • What does PCR stand for? • The Polymerase Chain Reaction.
4 - $200 • How many cycles of PCR is needed to make two molecules of your gene of interest? • Three
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
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.
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
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.
5 - $200 • What is the charge of DNA? • Negative
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.
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.
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.
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).