Open AccessA Simplified Method for the Determination of the Nucleotide Composition of Polyribonucleotides by Spectrophotometric Analysis
Author(s) -
RICHARDS E. G.
Publication year - 1968
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1968.tb00202.x
Subject(s) - hydrolysate , allowance (engineering) , chemistry , spectrophotometry , wavelength , composition (language) , analytical chemistry (journal) , component (thermodynamics) , fraction (chemistry) , nucleotide , spectrum (functional analysis) , chromatography , base (topology) , optics , mathematics , thermodynamics , hydrolysis , physics , biochemistry , mathematical analysis , quantum mechanics , mechanical engineering , linguistics , philosophy , engineering , gene
A simple method for detemining the base composition of ribonucleic acid hydrolysates is described. The method depends on the measurement of a difference spectrum between an aliquot of the hydrolysate at pH 7 and one at pH 11.8. This difference spectrum depends only on the concentrations of UMP and GMP present. The quantities of CMP and AMP are obtained from a spectrum at pH 7 after allowance for the contribution of the UMP and GMP. It is demonstrated that the results obtained by calculating the concentration of a component from measurements of absorbances at two wavelengths only are of comparable accuracy to those obtained from the more conventional technique of employing measurements at many wavelengths. It is concluded that the use of a digital computor is not required for the application of this method. A theoretical explanation is offered for this observation. The accuracy obtained is of the order of 0.02 in the mole fraction of a component.