Possible examination questions

CH329: Basics of Biotechnology

Lecture 1

1. Define bio-technology
2. A word bio-technology is a combination of two words. Explain each one briefly
3. Central dogma of molecular biology is made up of three components. Explain each one
briefly
4. Define (1) DNA replication (2) Transcription (3) Translation
5. Transcription and translation processes are much faster in prokaryotes cells than
eukaryotic cells. Explain
6. Give and explain one major roles of DNA and RNA
7. Draw the structure of Deoxyribose and Ribose
8. Explain briefly one mRNA can lead to the synthesis of multiple of proteins
9. What is the role of (1) codon? (2) anticodon?. Where will you find these codons and
anticodons
10. Given DNA sequence 5'GCGATATCGCAAA 3‘Write (1) mRNA (2) proteins sequence
using genetic code

Lecture 2

1. Define an enzyme and explain why enzymes are added in biochemical reactions
2. Three enzymes have permitted the development of various techniques used in analysis
and manipulation of DNA. Identify and explain the role of each enzyme
3. Both restriction enzymes and Dnases can break DNA. Why molecular biologist use
restriction enzymes during molecular cloning experiments
4. Both restriction enzymes and Dnases can break DNA. Why molecular biologist use
DNase during purification of RNA from DNA
5. What are the function of Dnase and Rnase during DNA and RNA purification?
6. Using the sequence 5'-GCGATA-3' draw DNA strands that will be formed after (1)
Blunt end digestion and (2) Staggered digestion
7. Restrictions enzymes that break DNA using staggered digestion are more useful to
molecular biologist than Blunt end digestion. Explain why
8. Define Recombinant DNA
9. What is the use of DNA ligase?
10. Briefly explain how CRISPR-Cas9 system is used in gene editing
11. Explain why Taq polymerase is used in PCR reaction than polymerase from E. coli
12. Define polymerase chain reaction (PCR) and explain how it works
13. Polymerase chain reaction (PCR) contains three stages (1) Denaturation (2) Annealing
(3) Extension. Explain the role of each stage briefly
14. What are primers and why are they added in PCR reaction?
 15. PCR reaction is used to make copies of DNA. Two factors determine the length of DNA
synthesized. Identify them and explain both of them
16. Draw an example of (1) good primer and (2) bad primer. What make a primer to be good
or bad?
17. Explain why a primer than is 20 base pair long is better than a primer than is 4 base pair
long
18. Explain the importance of doing the following steps in DNA purification (1)
homogenizing cells (2) doing cellular lysing (3) Adding chelating agents such as EDTA
(4) Adding protease K agents (4) phenol–chloroform mixture and (5) 100% ethanol
19. Given than the absorbance of DNA and RNA solutions are 0.245 and 0.0125. Calculate
the concentration of DNA and RNA in μg/ml
20. Explain how Agarose Gel electrophoresis is used to separate DNA fragments
21. How ethidium bromide is used in identification of DNA in Agarose Gel electrophoresis
22. What is Southern blot? Western blot? Northern blot?
23. What is DNA probe? How is it used in detecting gene of interest?
24. What is the purpose of Sanger sequencing?
25. What is the role of ddNTPs and dNTPs in Sanger sequencing?

Lecture 3
1. Define (1) Molecular Cloning and (2) Protein Expression (3) Molecular cloning
2. What are plasmids and why are they important in biotechnology?
3. There are five essential Essential Characteristics of Cloning Vectors. Explain each one
briefly
4. There are 7 basic steps involved in gene cloning. Explain each one briefly
5. Define (1) Transformation and (2) Competence and explain why each one is important in
gene cloning
6. What is Reporter gene and why is it important in Molecular cloning?
7. Define Gene induction.
8. An operon system is made up of 4 components: Explain each one briefly