plasmid subcloning - (Dec/04/2007 )
I am new to cloning. i am trying to add IRES sequence to plasmid containing RFP. The other plasmid I have is CRE-IRES-TK. So how do i make IRES- RFP. I have no primers. Do i basically cut both with respective enzymes and then ligated them?
If there are restriction sites on both the IRES vector and RFP, i would use them for cloning. As you wrote, cut them with the same enzymes, and ligate them.
PCR would be an option if there are no compatible sites on both plasmids.