A simple and effective strategy for preparing fluorophore-labelled PNA is described. A C-terminal S-t-butylmercaptocysteine-derivatized PNA was prepared on solid-phase using Fmoc chemistry. Selective deprotection of the S-t-butylmercapto group on-bead, allowed the free thiol to be reacted with a fluorophore derivatized via an iodoacetamido or maleimido linker. Subsequent cleavage and sidechain deprotection yielded C-terminal labelled PNA in good yield and purity. Dual labelled PNA was also prepared by using both C-terminal (-SH) and N-terminal (-NH(2)) labelling chemistries.