I have a plasmid which differs by one base from wild type. This base will change the protein by one amino acid, hence I need to mutate that base which is residing in the central part of the plasmid. Is inverse PCR the only option for it?
Now about designing the primer for inverse PCR- 1) Where should I put the mutated base - terminal or central position of the primer?
2) Should I put the mutated base in both forward and reverse primers or either of them?
Please illustrate with an example if possible.
Thanks