5-Ethynyl-dUTP (5-EdUTP)

Incorporation into DNA by PCR with family B polymerases (Pwo, Deep Vent exo- or KOD XL)[1,2] Incorporation into DNA by Primer Extension with family A polymerase Taq and family B polymerases (Pwo, Deep Vent exo- or KOD XL)[1] The resulting ethynyl-functionalized DNA can subsequently be processed via Cu(I)-catalyzed click chemistry that offers the choice to introduce a Biotin group (via Azides of Biotin) for subsequent purification tasks to introduce fluorescent group (via Azides of fluorescent dyes) for subsequent microscopic imaging to crosslink the DNA to Azide-functionalized biomolecules e.g.proteins Presolski et al.[3] and Hong et al.[4] provide a general protocol for Cu(I)-catalyzed click chemistry reactions that may be used as a starting point for the set up and optimization of individual assays.