Question

There is a plasmid containing eGFP on your workbench.

How can we cut the eGFP gene from the plasmid?
a) Insert the plasmid into bacteria so they secrete the gene
b) Perform restriction enzyme digestion
c) Isolate the DNA using phenol-chloroform extraction
d) Perform DNA sequencing on the whole plasmid

Answer 1

B) Perform Restriction Enzyme Digestion

Step-by-step explanation:

Restriction enzymes are used to cut DNA at specific locations, specific to each enzyme. Plasmids are double stranded pieces of DNA. In your case it encodes for a Green Fluorescent Protein. In order to cut out the GFP DNA or insert it into the plasmid it must first be "cut" with a restriction enzyme.

Attached is an image of a plasmid map. All of the lines that stick out from the circle represent sites where known restriction enzymes will cut. The label is the name of the enzyme. These enzymes cut at specific sequences that are typically palindromic.