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2. Cloning Experiments are the source of the word "recombinant" in Recombinant DNA Technology. Explain why.
3. Why is this difficult to do with mammalian genomes?
4. What is a "complementation assay" is the cloning, for example, of a bacterial gene?
5. What is Reverse Genetics? What are the steps in a Reverse Genetics approach to cloning a gene?
6. Once a gene is isolated via cloning, how would you proceed in analysis of the cloned DNA?
7. What is a cDNA?
8. What is cDNA cloning?
9. Why are four sequencing reactions performed in Sanger sequencing?

10. What is a Sequencing Gel? How does it differ from a standard R.fragment agarose gel?

11. What types of mutants would you isolate? Why?

12. What three major classes of E. coli dna mutants have been isolated? Which step in DNA

replication would you expect the gene products of each of these three gene classes to be

involved?

13. How have the E. coli dna mutants been used to purify DNA replication proteins?

14. What features of a Type II R.enzyme make possible cloning experiments?

15. What is a Restriction Enzyme?

16. What features of a Type II Restriction Enzyme are important for recombinant DNA work?
17.What is a "sticky end", and why is it said that some R.Enzymes generate "sticky ends"?

18. Why is this enzyme called a "6 cutter"?
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