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4′-SelenoDNA fragments were synthesized for the first time using 4′-selenothymidine triphosphate (SeTTP) by taking advantage of its bioequivalence against DNA polymerases. DNA fragments each with a homologous element (O, S or Se) at the 4′-position of the thymidine units were effectively amplified using KOD Dash DNA polymerase.

Graphical abstract: Synthesis of DNA fragments containing 2′-deoxy-4′-selenonucleoside units using DNA polymerases: comparison of dNTPs with O, S and Se at the 4′-position in replication

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