Question Bank – Recombinant DNA

Technology (Units I–III)

Unit I: Introduction to Recombinant DNA Technology
1. Differentiate between isoschizomers and neoschizomers with suitable examples.

2. Explain why star activity is a limitation in restriction enzyme use. Suggest ways to
minimize it.

3. MCQ: Which enzyme is used to remove 5’-phosphates from DNA ends to prevent self-
ligation?

4. One-word: Name the enzyme that can add nucleotides to the 3’ end of DNA in a template-
independent manner.

5. Why are cohesive ends preferred over blunt ends in cloning experiments?

6. Give reason: Why do restriction enzymes always cut at palindromic sequences?

7. True/False: Linkers and adaptors are the same. Justify your answer.

8. Match the following: (i) Polynucleotide kinase (ii) DNA ligase (iii) Alkaline phosphatase
(iv) Exonuclease → (a) Removes terminal phosphates (b) Joins DNA ends (c) Adds
phosphate group (d) Degrades from ends.

9. Describe the role of terminal transferase in constructing recombinant DNA molecules.

10. Short Answer: Outline the steps for plasmid isolation from bacteria.

11. MCQ: Which of the following enzymes is most commonly used for DNA polymerization
in rDNA technology?

12. Explain the concept of compatible cohesive ends with an example.

13. Why are isoschizomers important in designing cloning strategies?

14. One-word: Which enzyme fills in recessed 3’ ends of DNA?

15. Explain how alkaline phosphatase prevents unwanted religation.

16. Short Answer: Differentiate between inter- and intramolecular ligation with examples.

17. Why are linkers and adaptors used in rDNA experiments?

18. True/False: Star activity occurs due to optimal Mg2+ concentration. Justify.
19. What is restriction mapping? State its applications.

20. MCQ: Which type of restriction enzyme is widely used in cloning?

21. Why is ligase essential in recombinant DNA construction?

22. One-word: Enzyme that phosphorylates DNA ends?

23. What is the biological importance of restriction-modification systems in bacteria?

24. Short Answer: Explain the difference between blunt-end and sticky-end ligation
efficiency.

25. Give reason: Why is plasmid DNA isolation preferred over genomic DNA in cloning
experiments?

Unit II: Cloning Vectors

26. Differentiate between cloning and expression vectors.

27. MCQ: Which selection marker is used in blue-white screening?

28. One-word: What is the full form of MCS?

29. Why is host range an important consideration in vector selection?

30. Short Answer: Explain the principle of TA cloning.

31. Differentiate between transformation, transfection, and transduction.

32. What is a shuttle vector? Give one example.

33. Explain the role of copy number regulation in plasmids.

34. Give reason: Why are plasmid compatibility groups important in co-transformation?

35. True/False: Expression vectors always contain a promoter upstream of the gene of
interest. Justify.

36. MCQ: Which phage vector uses Spi⁻ selection?

37. What is in vitro packaging in λ DNA vectors?

38. Explain the difference between insertional and replacement λ vectors.

39. One-word: Name a bacteriophage-derived expression system widely used in E. coli.

40. What is the application of T7 expression system in biotechnology?

41. Match the following: (i) Cosmid (ii) Phagemid (iii) BAC (iv) YAC → (a) >100 kb inserts in
yeast (b) f1 origin + plasmid ori (c) 35–45 kb packaging (d) >100 kb inserts in bacteria.

42. Explain why artificial chromosomes are necessary for large DNA cloning.

43. Short Answer: How does blue-white screening distinguish recombinants?

44. Give reason: Why is competent cell preparation essential for transformation?

45. What are selection and screening markers? Differentiate with examples.

46. One-word: Which enzyme is exploited in blue-white screening?

47. True/False: Shuttle vectors can replicate in both prokaryotic and eukaryotic hosts.
Justify.

48. Why are λ vectors preferred for constructing genomic libraries?

49. MCQ: Which vector can accommodate inserts >300 kb?

50. Explain the advantages of phagemids in recombinant DNA technology.

Unit III: DNA Libraries

51. Explain the principle of genomic library construction.

52. Differentiate between genomic and cDNA libraries.

53. MCQ: Which enzyme is used to synthesize cDNA from mRNA?

54. One-word: What is chromosomal walking?

55. What are the limitations of cDNA library construction?

56. Give reason: Why is oligo(dT) primer used in cDNA synthesis?

57. Short Answer: Outline the steps in constructing a full-length cDNA library.

58. Explain the importance of differential cDNA libraries.

59. What is subtraction in library construction?

60. MCQ: Which method is used for analyzing differential expression of mRNA?

61. True/False: cDNA libraries contain introns. Justify.

62. Explain the use of degenerate primers in PCR-based library screening.

63. One-word: Technique to study protein-protein interaction in libraries?

64. Describe the principle of Southwestern screening.

65. Differentiate between Northwestern and Southwestern blotting.

66. Explain functional complementation in library screening.

67. Short Answer: Describe difference cloning.

68. MCQ: Which method uses labeled antibodies to detect proteins in libraries?

69. What is differential display PCR?

70. Give reason: Why is chromosomal walking essential in genome projects?

71. One-word: Which enzyme is essential for RT-PCR?

72. True/False: Genomic libraries represent only expressed genes. Justify.

73. What is the role of adaptors in library construction?

74. Explain immunochemical screening of cDNA libraries.

75. Match the following: (i) Genomic library (ii) cDNA library (iii) Subtracted cDNA library
(iv) Differential display → (a) Eliminates common transcripts (b) Represents whole genome
(c) Studies expression variation (d) Represents expressed genes only.

76. Differentiate between library screening by hybridization and PCR.

77. Short Answer: Outline the steps in constructing a subtracted cDNA library.

78. Give reason: Why is full-length cDNA difficult to construct?

79. Explain ligand-interaction-based screening of libraries.

80. MCQ: Which technique can detect gain-of-function clones?

81. What is meant by functional complementation approach?

82. One-word: Which type of blotting identifies RNA-binding proteins?

83. Describe how degenerate probes are designed for library screening.

84. True/False: Library screening using hybridization requires labeled DNA probes. Justify.

85. Explain the application of differential cDNA libraries in cancer research.

86. What is difference cloning?

87. MCQ: Which type of library is most suitable for identifying regulatory elements?

88. Give reason: Why are overlapping fragments important in genomic libraries?

89. Short Answer: Discuss PCR-based methods for library screening.

90. Describe how gain-of-function approaches are applied in library analysis.

91. One-word: Method used to identify DNA-binding proteins?

92. Explain subtracted cDNA libraries with one biological example.

93. True/False: Differential display PCR can identify both upregulated and downregulated
genes. Justify.

94. Differentiate between normalized and subtracted cDNA libraries.

95. Short Answer: Outline the principle of PCR-based differential display.

96. MCQ: Which approach can reveal unknown gene transcripts?

97. Explain the use of chromosomal walking in positional cloning.

98. Give reason: Why are protein-protein interaction screens essential in functional
genomics?

99. One-word: What is the main advantage of cDNA over genomic libraries in expressing
eukaryotic genes?

100. Describe the importance of gain-of-function library screens in biotechnology.