The typical click chemistry reaction is the Copper-catalyzed reaction of an azide with an alkyne to form a 5-membered heteroatom ring, also known as Cu(I)-Catalyzed Azide-Alkyne Cycloaddition (CuAAC). The azide and alkyne functional groups must first be introduced into the biomolecules to be linked. Alkyne-containing DNA can be prepared with an alkyne phosphoramidite during standard oligo synthesis. Azide or alkyne groups can be attached to proteins by using bifunctional adapter molecules with an NHS ester moiety. Aqueous DMSO, DMF, acetonitrile, alcohols, or pure water and buffers can be used for the reaction. The reaction is biocompatible and can take place in living cells.
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