RAPID AND SENSITIVE HIGH-PRESSURE LIQUID-CHROMATOGRAPHY ASSAY FOR DEOXYRIBONUCLEOSIDE TRIPHOSPHATES IN CELL-EXTRACTS

被引:285
作者
GARRETT, C [1 ]
SANTI, DV [1 ]
机构
[1] UNIV CALIF SAN FRANCISCO,DEPT PHARMACEUT CHEM,SAN FRANCISCO,CA 94143
基金
美国国家卫生研究院;
关键词
D O I
10.1016/S0003-2697(79)80005-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An assay for simultaneous quantitation of deoxyribonucleoside triphosphates (dNTPs) in cell extracts is described. Following destruction of ribonucleotides by treatment with periodate and methylamine, dNTPs are separated by high pressure liquid chromatography and quantitated by their ultraviolet absorbance upon elution. Destruction of ribonucleotides is complete (>99.9%), the resulting mixture is suitable for direct injection without further manipulation, and recovery of dNTPs is reproducible and near quantitative. This technique offers a simple and rapid alternative to either DNA polymerase or thin-layer chromatography assays for dNTPs in cellular extracts. © 1979 Academic Press, Inc.
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页码:268 / 273
页数:6
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