Highly fluorescent 5-(5,6-dimethoxybenzothiazol-2-yl)-2'-deoxyuridine 5'-triphosphate as an efficient substrate for DNA polymerases.

Highly fluorescent 5-(5,6-dimethoxybenzothiazol-2-yl)-2'-deoxyuridine 5'-triphosphate as an efficient substrate for DNA polymerases.

Chembiochem : a European journal of chemical biology (2011-09-03)

Kousuke Sato, Ayano Sasaki, Akira Matsuda

ABSTRACT

We herein describe the synthesis of fluorescent 5-(5,6-dimethoxybenzothiazol-2-yl)-2'-deoxyuridine 5'-triphosphate (d(bt)UTP) and primer extension reactions using d(bt)UTP. We also carried out primer extension reactions using the (bt)U template. B family DNA polymerases, such as KOD, Deep Vent (exo-), and 9°N(m) DNA polymerases, were effective for elongation with d(bt)UTP. Deep Vent (exo-) and KOD DNA polymerases have excellent fidelity for incorporating d(bt)UTP only at the site opposite the adenine template and only dATP when using the (bt)U template. Therefore, d(bt)UTP is an excellent fluorescent nucleotide that can be incorporated into DNA by DNA polymerases.

MATERIALS

Product Number

Brand

Product Description

D6920

Sigma-Aldrich

2′-Deoxyadenosine 5′-triphosphate sodium salt solution, 10 mM

D4788

Sigma-Aldrich

2′-Deoxyadenosine 5′-triphosphate sodium salt solution, 100 mM, pH 7

D6500

Sigma-Aldrich

2′-Deoxyadenosine 5′-triphosphate disodium salt, ≥97%