<article> With their broad chemical and functional diversity ncAAs provide a variety of promising applications including proteinlabeling, photocaging, structure analysis and specific protein interactions. Therefore, the development of a new iGEM toolkit for the translational incorporation of non-canonical amino acids (ncAAs) in E. coli would be a great contribution to the iGEM community. The site-specific incorporation of ncAAs requires the directed evolution of tRNA and aminoacyl tRNA synthetase pairs, which in turn mediate the introduction of a ncAA for a certain codon. We are especially exploring the application of unnatural base pairs (UBPs) as an expansion of the genetic code. This approach promises an optimal orthogonality to the autologous translation apparatus and a high flexibility concerning the incorporation of multiple ncAAs. We will utilize several systems to achieve a high retention efficiency. As examples for the successful incorporation of non-canonical amino acids we will develop a rapid test for prions and a new chromatography method for a mild elution of proteins.</article>