5-Ethynyl-dUTP (5-EdUTP) is easily incorporated into DNA by PCR instead of dUTP using polymerases like Pwo, Deep Vent exo- or KOD XL. The resulting stable and modified PCR fragment is now ready for further functionalization by Click Chemistry. We recommend a substitution of 1 – 10% of dUTP.

LITERATURE

Synthesis of Highly Modified DNA by a Combination of PCR with Alkyne-Bearing Triphosphates and Click Chemistry, J. Gierlich et al., 2007, Chem. Eur. J., Vol. 13(34), p. 9486–9494.

https://doi.org/10.1002/chem.200700502

Directed DNA Metallization, G. A. Burley et al., 2006, J. Am. Chem. Soc., Vol. 128(5), p. 1398–1399.

https://doi.org/10.1021/ja055517v

Identification and characterization of nucleobase-modified aptamers by click-SELEX, F. Pfeiffer et al., 2018, Nature Protocols, Vol. 13(5), p. 1153–1180.

https://doi.org/10.1038/nprot.2018.023

Quantitation of deoxynucleoside triphosphates by click reactions, C. Y. Huang et al., 2020, Scientific Reports, Vol. 10(1), p. 611.

https://doi.org/10.1038/s41598-020-57463-3