Precision and accuracy of anion-exchange separation of nucleic acids
OwnerWill Blochinventor
Life of this patent
Estimating the term…
Abstract
Solvents for salt-gradient anion-exchange separation of nucleic acids, especially double-stranded DNA and especially by liquid chromatography, are improved by replacing NaCl as the eluting salt with any of a wide range of alkyl ammonium salts and can be used to elute nucleic acids in strict order of increasing length, with reduced sensitivity to elution temperature and salt concentration. Anion-exchange chromatography with these solvents is well suited for identification of DNA fragments on the basis of size, with greater accuracy, precision, and resolvable size range than often is possible with gel electrophoresis.
Claims9
A process for separating and analyzing double stranded deoxyribonucleic acids on the basis of differences in molecular size, said process comprising:
- (a) contacting a test sample containing double stranded deoxyribonucleic acid with an anion-exchange solid;
- (b) contacting said solid of step (a) with a series of solvents possessing a pH in the range of 4 to 9 comprising:
- an eluting salt in the concentration range of 0.5M to 1.5M composed of equal concentrations of:
- a cation selected from the group consisting of dialkylammonium, trialkylammonium and tetraalkylammonium wherein the alkyl groups consist of any combination of methyl and ethyl;
- and
- an anion selected from the group consisting of bromide, chloride, acetate, formate;
- nitrate, perchlorate, dihydrogen phosphate, ethane sulfonate, and methane sulfonate;
- and
- a buffer acid with a pKa in the approximate range of 3.5 to 9.5;
- wherein the concentration of eluting salt is systematically increased from approximately 0.5M to approximately 1.5M such that a major portion of the series of solvents is separated from the solid after said contacting;
- and
- (c) analyzing the series of solvents for the double stranded deoxyribonucleic acid.
Dependencies are inferred from the claim text.