Acetate-free purification of plasmid dna on hydroxyapatite

Details Acetate-free purification of plasmid dna on hydroxyapatite Acetate-free purification of plasmid dna on hydroxyapatite

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01) B01D 15/36 (2006.01)

Patent

CA 2414788

Plasmid DNA is purified from a cell lysate by acidifying the lysate with a combination of a mineral acid and an inorganic salt in place of the commonly used acetate buffer, allowing chromosomal DNA and other impurities to precipitate out and recovering the supernatant, and applying the supernatant directly to a hydroxyapatite chromatographic separation medium. The use of the mineral acid and organic salt avoids the degradation of the hydroxyapatite that frequently accompanies repeated use with acetate and related compounds.

De l'ADN plasmidique est purifié à partir d'un lysat cellulaire par acidification du lysat avec une combinaison d'un acide minéral et d'un sel inorganique à la place du tampon d'acétate habituellement utilisé, par formation d'un précipité d'ADN chromosomique et d'impuretés et récupération du surnageant, et par application du surnageant directement sur un milieu de séparation chromatographique d'hydroxyapatite. L'utilisation de l'acide minéral et de l'acide organique évite la dégradation de l'hydroxyapatite accompagnant fréquemment l'utilisation répétée avec des composés d'acétate et apparentés.

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