Aiming to bypass the individual purification of the amino acid tRNA synthetases as well as the individual tRNAs, we tested a bacterial lysate (S30 extract) isolated from E. coli containing amino acids, tRNA and amino acid tRNA synthetases. Although initially easy to produce, this S30 extract was not as efficient at yielding CDPs as expected (Fig. ). Furthermore, use of noncanonical amino acids with the S30 extract was unsuccessful due to the high levels of endogenous aminoacylated tRNA present.