Recombinant DNA Technology
01. What is an episome. What are plasmids.
– What are the natural function of a bacterial plasmid
– What is the difference between a plasmid and a virus (phage)
– What is conjugation. How it is accomplished and what bacterial structures are required for the process. What is the outcome of conjugation.
02. Plasmid structure
– What are the elements for a functional plasmid as a genetic tool
– What are the advantages of plasmids as genetic tools
03. Restriction endonucleases and restriction sites
– What are the sequence requirements for a restriction site. What does palindromic is. How is it read on DNA
– How the bacteria keeps its DNA from being cut by its own restriction endonucleases. What is a DNA methylase.
– Sticky end vs blunt ends
– What is the frequency of a 4 nt-long restriction site. How about an 8nt-long? What about a 20-nt long? How do you determine it?
04. Cloning genes into plasmids.
– How does the lacZ gene serve as a screening tool for cloning genes into plasmids.
– What is X-Gal and what do you use it for. The white/blue colony screening
– What is transformation
– How do you select for bacteria that have a plasmid inside
05. DNA gel electrophoresis
– The electrophoresis components
– Where does DNA migrate to and how do you detect it. How does Ethidium bormide work.
-How does supercoiled and relaxed DNA run in the gel
