Method for obtaining plasmid-dna by means of an aqueous biphasic system

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Method for obtaining plasmid-dna by means of an aqueous biphasic system
Agent: Leon R Yankwich Yankwich & Associates - Cambridge, MA, US
Inventors: Markus Muller, Jurgen Hubbuch, Andreas Frerix, Maria-Regina Kula
Applicaton #: 20060286080
Class: 424093210 (USPTO)
12/21/06
The invention relates to a process for isolation of plasmid DNA from biomass by means of an aqueous 2-phase system having a polymer component and a salt component, characterized in that the resuspension of the biomass employed, the alkaline lysis of the biomass, the neutralization of the alkaline lysis batch and the separation of the plasmid DNA from the contaminants (such as e.g. cell debris, RNA and gDNA) are carried out in a single reaction vessel (one-pot process). According to the invention, this is rendered possible in that the neutralization of the alkaline lysis batch is carried out in one and the same container by addition of potassium phosphate and one component of the aqueous 2-phase system is therefore already present, and in that the second component of the aqueous 2-phase system is a PEG having a molecular weight of the mathematical average of about 600 g/mol to 1,000 g/mol, but is preferably formed from a mixture of PEG 600 and PEG 1000.

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