反应详情

To a solution of 2 mg of 5-(3-aminoallyl)-2′-deoxyuridine-5′-triphosphate, ammonium salt (Sigma Chemical) in 100 μL of water is added a solution of 3 mg of Compound 72 (Example 73) in 100 μL of DMF, followed by addition of 5 μL of triethylamine. After the mixture is stirred at room temperature for 3 hours, the solution is evaporated to dryness under vacuum and the residue is purified by chromatography over lipophilic SEPHADEX resin using water for elution. The first green fluorescent fractions are combined and lyophilized to give the fluorescent nucleotide conjugate as an orange solid (Compound 107) Alternatively a fluorescent conjugate (Compound 108) of deoxyuridine-5′-triphosphate is prepared using 5-(3-amino-1-propynyl)-2′-deoxyuridine-5′-triphosphate in place of 5-(3-aminoally)-2′-deoxyuridine-5′-triphosphate (as described in Hobbs, Jr. et al, supra).

1. customthe solution is evaporated to dryness under vacuum
2. customthe residue is purified by chromatography over lipophilic SEPHADEX resin
3. washfor elution