Question

Describe a strategy using restriction enzymes and gel electrophoresis that would allow you identify that you have obtained the Recombinant DNA plasmid (containing YFG). Be specific indicating the techniques you would use and expected results.

Answer 1

Use a step wise process

Step-by-step explanation:

In order to find the presence of the recombinant protein:

1. Culture the cells that you believe harbors the plasmid for the recombination protein (+) and also cells that do not (-). This is your control.
2. Extract the plasmid from both strains.
3. Observe the plasmid map and the area of your insert. Use this to select the restriction enzymes at the beginning and the end of the YFG gene. You can also use a restriction site inside the gene but not in the plasmid for better control.
4. Perform the restriction enzyme digest on both (+) and (-) according to your protocol and separate using gel electrophoresis.
5. Observe the separate bands that you see. In the (+) there should be a band that is the size of your insert and a larger band that is the size of your plasmid control band and in the (-) there should be a single band that is the size of the plasmid but non the size of the insert.