How plasmids are used in recombinant DNA technology by illustrating the steps in constructing a plasmid designed to produce human growth factor in bacteria.
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1. Isolation of the Genetic Material (DNA)
2. Cutting of DNA at Specific Locations
Restriction enzyme digestions are performed by incubating purified DNA molecules with the restriction enzyme.
3. Isolation of Desired DNA Fragment
Using agarose gel electrophoresis, the activity of the restriction enzymes can be checked. Since, the DNA is negatively charged, it moves towards the positive electrode or anode and DNA tends to separates out in this process. After that the desired DNA fragment is eluted 4. Amplification of Gene of Interest using PCR
2. Cutting of DNA at Specific Locations
Restriction enzyme digestions are performed by incubating purified DNA molecules with the restriction enzyme.
3. Isolation of Desired DNA Fragment
Using agarose gel electrophoresis, the activity of the restriction enzymes can be checked. Since, the DNA is negatively charged, it moves towards the positive electrode or anode and DNA tends to separates out in this process. After that the desired DNA fragment is eluted 4. Amplification of Gene of Interest using PCR
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