Method for rnase- and organic solvent-free plasmid dna...

Details Method for rnase- and organic solvent-free plasmid dna... Method for rnase- and organic solvent-free plasmid dna...

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01)

Patent

CA 2378944

A process is described for purifying plasmid DNA from prokaryotic cells comprised thereof. This process comprises the steps of: (a) digesting the cells; (b) incubating the cells for about 4 to 24 hours to effect lysis and solubilization thereof, without effecting enzymatic digestion of RNA; (c) removing lysate contaminants from the cells to provide a plasmid DNA solution; (d) filtering the solution through a tangential flow filtration device to obtain a retentate containing the plasmid DNA; and (e) collecting the retentate.

L'invention concerne un procédé servant à purifier de l'ADN de plasmide depuis des cellules procaryotes. Ce procédé consiste à: (a) effectuer la digestion des cellules; (b) effectuer l'incubation de ces cellules pendant 4 à 24 heures afin d'en obtenir la lyse et la solubilisation sans effectuer de digestion enzymatique d'ARN; (c) supprimer des cellules les contaminants du lysat afin d'obtenir une solution d'ADN de plasmide; (d) filtrer la solution au moyen d'un dispositif de filtration d'écoulement tangentiel, de manière à obtenir un rétentat contenant l'ADN de plasmide; (e) recueillir le rétentat.

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