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Europium Cryptate Labeled Deoxyuridine-Triphosphate Analog: Synthesis and Enzymatic Incorporation

2000; Taylor & Francis; Volume: 19; Issue: 9 Linguagem: Inglês

10.1080/15257770008033854

1532-2335

Béatrice Alpha‐Bazin, Hervé Bazin, Sabrina Guillemer, Sylvie Sauvaigo, Gérard Mathis,

Molecular Biology Techniques and Applications

The synthesis of an europium tris-bipyridine cryptate labeled 2'-deoxyuridine-5 '-triphosphate analog (K-11-dUTP) is described. This labeled triphosphate was incorporated into DNA through enzymatic reactions with terminal transferase and DNA polymerases. The enzymatic reactions were monitored by TRACE (Time Resolved Amplification of Cryptate Emission), a homogeneous method using Fluorescence Resonance Energy Transfer (FRET) from an europium cryptate as donor to a modified allophycocyanine as acceptor.