MBB501- Principles of Biotechnology Question Paper

Mid-term Examination (2019-20): Principles of Biotechnology (MBB501)
Total 40 marks; Time: 2 hr

1. Answer the following questions (any five) in brief.

a) How bacterial cells protect its own DNA from the endogenous restriction enzymes.
(5x4= 20 marks)

b) What is alpha complementation? How it has been used for screening of recombinants in pUC18?

c) Write the components of reaction mixture with tentative concentration of each component in Polymerase chain reaction (PCR). How can you minimize PCR generated mutations (error) while amplifying a gene of 5 kb length?

d) Why mostly type II restriction enzymes are used in gene cloning experiments?

e) Differentiate between phagemid and cosmid vectors (you may add a map of each type in your answer)?

f) What is the significance of Chargaff's rule in deducing double helix structure of DNA?

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2. Answer the following questions in brief. You may use suitable diagrams appropriately. (3x10= 30 marks)

A. What are the different mechanisms of selecting recombinants while cloning in lambda phage based vectors. Include examples of phage vector in case of each type.

B. Describe minimum two methods of manipulating DNA ends for conversion of blunt ends to sticky ends and vice versa.

C. What is a degenerate primer? What are the important guiding principles when a degenerate primer is designed from a single protein sequence and from a conserved domain of several homologous nucleotide sequences?

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