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Updated: Jun 15, 2026
Chemical Triphosphorylation of Oligonucleotides
Published on: June 2, 2022
Quantitative analysis of ribonucleotide triphosphates in cell extracts by high-performance liquid chromatography and
A Loregian1, C Scremin, M Schlavon
1Institute of Microbiology University of Padova, via A. Gabelli 63, 35121 Padova, Italy.
Abstract:
Two analytical methods have been evaluated for their ability to separate and quantitate the intracellular pool of free ribonucleotide triphosphates in four different cell lines. Micellar electrokinetic capillary chromatography and high performance liquid chromatography were compared in terms of speed, sensitivity, and efficiency of analysis, with the former being 3 times faster (about 10 vs 30 min) and having an average minimun detectable quantity of 50 fmol vs 30 pmol and approximately 950 000 theorical plates/m vs 4500. Different protocols of extraction of nucleotides from whole cells were also evaluated with the two analytical methods. Micellar electrokinetic capillary chromatography proved to be a powerful tool for the reproducible and reliable quantitation of ribonucleotide triphosphates, allowing the fast processing of extremely small volumes of sample.
