PMID: 3766933Aug 1, 1986Paper

Quantitative determination of antiviral nucleoside analog in DNA

Analytical Biochemistry
M S ChenS C Oshana

Abstract

A technique for the analysis of the amount of an antiviral nucleoside analog incorporated into DNA, utilizing enzymatic digestion of DNA, followed by high-performance liquid chromatography is described. The cells or tissue samples were treated with perchloric acid to inactivate the nucleases, then digested with pronase in the presence of EDTA. DNA was purified by CsCl centrifugation followed by Sephadex chromatography and treatment with deoxyribonuclease 1 and venom phosphodiesterase. The deoxyribonucleoside monophosphates and the monophosphate of the nucleoside analog liberated from DNA were separated and quantitated by HPLC analysis and measurement of radioactivity. This assay is more sensitive, specific, and precise than the determination of DNA density shift. It is also applicable for nucleoside analogs which do not change the density of DNA either because of their structure or their very small degree of incorporation.

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·G B ElionH J Schaeffer
Jun 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·E De ClercqR T Walker
Dec 1, 1967·Tetrahedron Letters·M YoshikawaT Takenishi

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Citations

Jul 10, 1998·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·B C TennantJ L Gerin
Jun 1, 1996·Toxicology in Vitro : an International Journal Published in Association with BIBRA·J M ColacinoF C Richardson
Dec 6, 1994·Proceedings of the National Academy of Sciences of the United States of America·F C RichardsonR R Bowsher

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