Acyclic nucleoside triphosphate analogs as terminators in biocatalytic DNA replication

Bioorganic and Medicinal Chemistry Letters

Volumn 7, Issue 23, 1997, Pages 3013-3016

  Martinez, Carlos I ^a   Ansari, M Ali ^b   Gibbs, Richard ^b   Burgess, Kevin ^a  

^a TEXAS A AND M UNIVERSITY   (United States)

^b BAYLOR COLLEGE OF MEDICINE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

ACYCLIC NUCLEOSIDE; DNA POLYMERASE;

ARTICLE; CATALYSIS; DNA REPLICATION; DRUG SYNTHESIS; ENZYME SUBSTRATE; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; HUMAN IMMUNODEFICIENCY VIRUS 1; IN VITRO STUDY; MASS SPECTROMETRY; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; REACTION ANALYSIS;

EID: 0343471106     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0960-894X(97)10135-4     Document Type: Article

References (8)

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7. 32P-labeled primer, melted at 80 °C, then allowed to anneal by slow cooling to room temperature. The double-strand DNA so produced was mixed with the analog (1 or 2) and the various dNTP and ddNTP's at the concentrations given above. After incubation for 15 min at the temperature prescribed for each enzyme, the reactions were quenched by addition of 5 μL of stop solution containing 98% diisopropyl formamide, 10 mM EDTA, pH 8.0, 0.025% bromophenol blue, 0.025% xylene cyanol. The samples were heated to 85 °C for 5 min, chilled in ice, and 3 μL were loaded into 20% acrylamide gels. Following electrophoresis the gel was fixed in aqueous 10% acetic acid, 10% methanolic solution (v/v), dried, and autoradiographed on Hyperfilm TM-MP (Amersham).
8. Tabor, S.; Richardson, C. C. Proc. Natl. Acad. Sci. U.S.A. 1995, 92, 6339.