1. Pseudomonas putia is thought to degrade hydrocarbons due to the fact that it harbours a TOL plasmid. To verify this assertion, a student was tasked to isolate and grow this soil-dwelling bacteria and to also isolate the plasmid from the microbe. The TOL plasmid is also intended to be used as a cloning vector.
A. Elucidate two physical properties that will enable you to isolate TOL plasmid from the chromosomal DNA of the bacteria.
B. Describe an appropriate method you could use to isolate TOL plasmid from the bacteria.
C. During the isolation of TOL plasmid from Pseudomonas putida, it was observed that the concentration of the plasmid DNA per 50 mL of sample was extremely low. Describe a technique you could employ to increase the concentration of the TOL plasmid.
D. Demonstrate a procedure that you could adopt to restrict or digest the TOL plasmid.
E. How would you ascertain if your restriction experiment was successful?
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