Synthesis of 3′-O-(ω-aminoalkoxymethyl)thymidine 5′-triphosphates, terminators of DNA synthesis that enable 3′-labelling
Abstract
Treatment of 5′-O-benzoylthymidine 1 with a mixture of acetic anhydride, acetic acid and dimethyl sulfoxide yielded 5′-O-benzoyl-3′-O-methylthiomethylthymidine 2, which was converted via the 3′O-bromomethyl derivative into 3′-O-(ω-aminoalkoxymethyl)thymidines 7 bearing a 6, 8 or 10 methylene groups long hydrocarbon chain, and finally to their 5′-triphosphates 10. The latter compounds were shown to be terminators of DNA synthesis catalysed by thermostable Tet/z DNA-polymerase, and may be labelled at the aliphatic amino group with fluorescent probes.