Short Answer TypePolymerase Chain Reaction or PCR is used to amplify the gene of interest. In this method -:
Primers are designed according to the sequence of the gene of interest.
Two sets of primers (chemically synthesized oligonucleotide stretches) that are complementary to the gene of interest, DNA polymerase enzyme, and deoxynucleotides are added to the PCR mixture.
PCR consists of 3 steps:
i. Denaturation The ds DNA is denatured by providing high temperature. Taq DNA polymerase isolated from the thermophilic bacteria, Thermus aquaticus (Taq), which does not degrade at such high temperatures is used.
(a) Name the selectable markers in the cloning vector pBR322 ? Mention the role they play.
(b) Why is the coding sequence of an enzyme 
-galactosidase a preferred selectable marker in comparison to the ones named above ?
Suggest and describe a technique to obtain multiple copies of a gene of interest in vitro.
Long Answer TypeA. Describe the characteristics a cloning vector must possess.
B. Why DNA cannot pass through the cell membrane? Explain. How is a bacterial cell made competent to take up recombinant DNA from the medium?
If a desired gene is identified in an organism for some experiments, explain the process of the following:
(i) Cutting this desired gene at specific location
(ii) Synthesis of multiple copies of this desired gene
Short Answer Type
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