I am currently trying to insert a gene of interest from a plasmid into a different empty plasmid. In order to cut the gene of interest from the plasmid, I know I have to use restriction enzymes. However, since the plasmid is circular, would I have to cut the plasmid with two different restriction enzymes or one restriction enzyme that cuts at two places? Also, if the restriction enzyme is supposed to match the enzyme I will be using for the empty plasmid, how is this possible if I use two different restriction enzymes for the plasmid containing my gene of interest?
I'm just confused about the details of this procedure because all of the research I have found online relates to a gene of interest that is not in a plasmid.
If anyone could help me, that would be great. Thanks!