Precision and accuracy of anion-exchange separation of nucleic acids
First Claim
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1. An aqueous solvent useful for the anion-exchange chromatography of double-stranded nucleic acids comprising:
- (a) an eluting salt in the approximate concentration range of 0.5 to 1.5M composed of equal concentrations of;
(1) a tetramethylammonium cation and(2) an anion selected form the group consisting of formate and nitrate, and(b) a buffer acid with a pKa in the approximate range of 3.5. to 9.5, which acid has a concentration not exceeding approximately 0.05M;
wherein the eluting salt and the buffer are components of an aqueous solution.
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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.
42 Citations
10 Claims
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1. An aqueous solvent useful for the anion-exchange chromatography of double-stranded nucleic acids comprising:
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(a) an eluting salt in the approximate concentration range of 0.5 to 1.5M composed of equal concentrations of; (1) a tetramethylammonium cation and (2) an anion selected form the group consisting of formate and nitrate, and (b) a buffer acid with a pKa in the approximate range of 3.5. to 9.5, which acid has a concentration not exceeding approximately 0.05M; wherein the eluting salt and the buffer are components of an aqueous solution. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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Specification
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Current AssigneeApplied Biosystems LLC (Thermo Fisher Scientific Incorporated)
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Original AssigneePerkinelmer Incorporated
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InventorsBloch, Will
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Primary Examiner(s)LUDLOW, JAN M
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Application NumberUS08/589,904Time in Patent Office1,078 DaysField of Search435/6, 536/26.42, 536/26.43, 536/25.4, 935/19, 935/20, 436/161, 436/18, 564/281, 564/291US Class Current436/18CPC Class CodesB01D 15/36 involving ionic interactionB01D 15/363 Anion-exchangeB01D 15/426 Specific type of solventB01J 41/20 Anion exchangers for chroma...G01N 2030/8813 biological materialsG01N 30/02 Column chromatographyG01N 30/34 of fluid composition, e.g. ...Y10T 436/108331 Preservative, buffer, antic...Y10T 436/143333 Saccharide [e.g., DNA, etc.]
