Measurement of deoxyguanosine/thymidine ratios in complex mixtures by high-performance liquid chromatography for determination of the mole percentage guanine + cytosine of DNA.
The ratio of deoxyguanosine and thymidine can be determined in a complex mixture containing the major ribonucleosides and deoxynucleosides, the minor deoxynucleosides, and the nucleotide monophosphates by high-performance liquid chromatography. The isocratic procedure utilizes a C18 column and a solvent of methanol-triethylamine phosphate (pH 5.1). A single analysis requires 15 min. Within the range of 0.5-1.5 micrograms of total deoxynucleosides per sample, the determination is very precise and the relative standard deviation is about 0.1%. From the deoxyguanosine/thymidine ratio, a precise determination of the mole percentage guanine + cytosine of double-stranded DNA is calculated.
Mesbah
M
M
Department of Microbiology, University of Georgia, Athens 30602.
Whitman
W B
WB
D001105:Archaea / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003596:Cytosine / Q000032:analysis*; D004247:DNA / Q000032:analysis*; D004269:DNA, Bacterial / Q000032:analysis; D003849:Deoxyguanosine / Q000032:analysis*; D006147:Guanine / Q000032:analysis*; D006868:Hydrolysis; D013056:Spectrophotometry, Ultraviolet; D013936:Thymidine / Q000032:analysis*
D004269:DNA, Bacterial; D006147:Guanine; D003596:Cytosine; D004247:DNA; D003849:Deoxyguanosine; D013936:Thymidine