Question 3: A recombinant DNA molecule was created by ligating a gene to a plasmid vector. By mistake, an exonuclease was added to the tube containing the recombinant DNA. How does this affect the next step in the experiment i.e. bacterial transformation?
Answer:
Recombinant DNA technology is a technique which enables the DNA to be identified, isolated and then recombined by which new features can be introduced into the organism. Recombinants are the hybrid DNA molecules which consists of self- replicating DNA fragments and inserted elements. These recombinants DNA molecules are ligated with desired DNA fragments to the replicon. This step is finalised by cutting the targeted DNA and a replicon with the restriction endonuclease enzyme which cuts the DNA at the specific position and it will lead to various identical copies of one cell or one fragment.
Hence, exonuclease will not affect recombinant- DNA because of its circular shape with no free ends. After making a circular DNA it will not affect because for making replication we need restriction endonuclease instead of exonuclease.
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