Because of their unique mechanism of cytotoxicity against bacteria and other microorganisms, antimicrobial peptides have received a great deal of attention as possible therapeutic agents. Incorporation of unnatural amino acids into the peptide sequences has the potential to improve the organism selectivity and potency of these peptides as well as increase their metabolic stability. This protocol outlines the logic used to selectively incorporate unnatural amino acid into a peptide sequence in an attempt to obtain peptides with increased therapeutic potential as antibiotic agents.