Nucleic acid purification using silica gel and glass particles

Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives

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435 911, C12P 1934, C07H 2104

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active

058080414

ABSTRACT:
The present invention provides compositions and methods for isolating nucleic acids with lengths greater than about 50 bases, from cells, gels, solutions and other media, in which nucleic acids occur in vivo or in vitro. The compositions of the invention are mixtures of the silica materials silica gel and glass particles, particularly glass microfibers; such mixtures combined with chaotropic salts, such as guanidinium chloride or guanidinium thiocyanate; and suspensions of such mixtures in aqueous solutions of chaotropic salts. In the methods of the invention, an aqueous solution comprising nucleic acid is mixed with an aqueous solution of chaotropic salts and the resulting solution is contacted with a mixture of the silica materials, whereupon the nucleic acid in the solution binds to the silica materials. The chaotropic salts and components, other than the nucleic acid adsorbed to the silica materials, from the aqueous solution treated by the method of the invention are washed from the silica materials. Finally, the nucleic acid can be obtained by elution from the silica materials. The methods provide nucleic acid in water or buffer, such as TE buffer, free of contamination by any salt or macromolecule that would interfere with further processing or analysis.

REFERENCES:
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Boom et al. J. Clin. Micro. 28:495-503, 1990.
Marko, et al., "A Procedure for the Large-Scale Isolation of Highly Purified Plasmid DNA Using Alkaline Extraction and Binding to Glass Powder", Analytical Biochemistry, vol. 121, pp. 382-387, 1982.
Chen, et al., "Recovery of DNA Segments form Agarose Gels", Analytical Biochemistry, vol. 101, pp. 339-341, 1980.
Vogelstein, et al., "Preparative and analytical purification of DNA from agarose", Proc. Natl. Acad. Sci. USA, vol. 76, No. 2, pp. 615-619, 1979.

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