Chapters 5 – Tutorial Questions

1. What is meant by the statement that DNA is replicated semiconservatively?

2. Explain the most important features of B-DNA.
3. Explain the molecular events of nucleic acid denaturation and renaturation.
4. Why is DNA denaturation a cooperative process?
5. Define the melting temperature of DNA
6. Why is the melting temperature of DNA with a higher fraction of G/C pairing, higher?
7. Compare RNA and DNA polymerases.
8. If RNA synthesis occurs in the 5’→3’ direction, in what direction is the DNA template
read?
9. Describe the function of the lac operon.
10. What is meant by an efficient/inefficient promoter?
11. Why is charging of a tRNA with the correct amino acid critical for correct translation?
12. In what direction does the ribosome read mRNA?
13. What factors contribute to the fidelity of DNA replication?
14. Explain the function of DNA ligase during DNA replication.
15. Explain the “restriction” and “modification” system in bacteria and the significance
thereof in the development of recombinant DNA technology?
16. What type of ends can be produced during restriction digestion?
17. How are bacterial cells made transformation competent?
18. Explain the basis for the select for recombinant clones using blue/white selection.
19. You want to clone a DNA fragment into a cloning vector, but the DNA fragment does
not contain any matching restriction enzyme sites with the cloning vector. Explain
how you can overcome this problem.
20. Why is a genomic library larger than a cDNA library for a given organism?
21. Explain the process of Southern Blotting.
22. How would you detect your gene of interest when you only have the amino acid
sequence available?
23. What is a degenerate probe?
24. What components does a PCR reaction typically contain?
25. Describe the different steps during a PCR reaction.
26. Why does Pfu DNA pol has a higher fidelity than Taq DNA pol?
27. What PCR-based method is commonly used in population studies through the
detection of a marker region?
28. Is it possible to amplify RNA using PCR? If so, how?
29. Explain the principle of a nested PCR.
30. During the recombinant production of proteins in bacteria one can encounter
various problems. List these problems and describe how they can be overcome.
31. Why is it problematic to express eukaryotic proteins in bacteria and can these
problems be overcome? How?
32. Through what process would you introduce a change in the amino acid sequence of
a protein using recombinant DNA technology?
33. What is the major difference between the reporter genes GFP and the lacZ gene?
34. Explain the different types of gene therapy.

Chapter 7 – Tutorial Questions

1. Explain automated Sanger sequencing.
2. What are the main steps to perform an NGS experiment?
3. Name the applications of NGS.
4. Explain the concept of “personalized medicine”.
5. Name some applications for DNA Microarrays.
6. What is the principle of DNA Microarrays?