Purification of nucleic acid from a sample containing nucleic acid and endotoxin
First Claim
1. A process for purifying nucleic acids from a biologic sample containing nucleic acid, comprising the following steps:
- a) preparing a fluid sample containing nucleic acid and endotoxin each in dissolved form;
b) precipitating at least the nucleic acid and endotoxin from the fluid sample onto the surface of a solid carrier material selected from the group consisting of mineral carrier materials, quartz fibers, silica, glass, aluminum oxide, zeolite, titanium dioxide, zirconium dioxide, and combinations thereof;
c) washing the precipitated constituents of the liquid sample from step b) to at least partially remove the endotoxin with a washing solution which contains(i) an amine compound of formula (I),
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Abstract
Disclosed is a process for purifying nucleic acids, especially plasmid DNA, from a nucleic acid-containing, biologic sample, consisting of (a) preparation of a fluid sample containing nucleic acid and endotoxin each in dissolved form; (b) precipitation at least of nucleic acid and endotoxin from the fluid sample; (c) washing of the components of the fluid sample precipitated out in step b) in order to at least partially remove the endotoxin with at least one washing solution, which contains at least one amine compound with at least two carbon atoms and with a molar mass of ≤500 g/mol and; at least one organic solvent different from the aforementioned amine compound and has a pH-value (20° C.) in the range from pH 3.0 to pH 8.5, and; (d) dissolution of the remaining nucleic acid from the washed, precipitated constituents from step c) using a dissolving buffer and collection of the dissolved nucleic acids in a separate receptacle. The process is suitable for the effective and economical removal of endotoxins.
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Citations
12 Claims
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1. A process for purifying nucleic acids from a biologic sample containing nucleic acid, comprising the following steps:
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a) preparing a fluid sample containing nucleic acid and endotoxin each in dissolved form; b) precipitating at least the nucleic acid and endotoxin from the fluid sample onto the surface of a solid carrier material selected from the group consisting of mineral carrier materials, quartz fibers, silica, glass, aluminum oxide, zeolite, titanium dioxide, zirconium dioxide, and combinations thereof; c) washing the precipitated constituents of the liquid sample from step b) to at least partially remove the endotoxin with a washing solution which contains (i) an amine compound of formula (I), - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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Specification