This unit contains detailed synthetic procedures for the implementation of the sulfo-click reaction to nucleoside derivatives. First, 3'-O-TBDMS-protected nucleosides are converted to their corresponding 4'-thioacide derivatives in three steps. Then, various conjugates are synthetized via a biocompatible and chemoselective coupling procedure using sulfonyl-azides partners. Finally, to illustrate the potential of the sulfo-click reaction a nucleoside bearing two orthogonal azido groups is synthetized and engaged in a one-pot dual labelling through a sulfo-click / copper-catalyzed azide−alkyne cycloaddition (CuAAC) cascade. The high efficiency of the sulfo-click reaction applied to nucleosides opens up new possibilities in the context of bioconjugation.