Highly Fluorescent 5-(5,6-Dimethoxybenzothiazol-2-yl)-2′-Deoxyuridine 5′-Triphosphate as an Efficient Substrate for DNA Polymerases

✍ Scribed by Dr. Kousuke Sato; Ayano Sasaki; Prof.Dr. Akira Matsuda

• Publisher: John Wiley and Sons
• Year: 2011
• Tongue: English
• Weight: 514 KB
• Volume: 12
• Category: Article
• ISSN: 1439-4227
• DOI: 10.1002/cbic.201100452

✦ Synopsis

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.