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US 6,548,253 B1Granted
Method of isolating plasmid DNA
AssigneeMerck Patent GmbH
InventorsKarl Holschuh, Uwe Michelsen
Life of this patent
Estimating the term…
Abstract
The invention relates to a method of isolating plasmid DNA from microorganism cultures with the aid of solid-phase bodies. The solid-phase bodies can be silica gels, silicates, or glass-like materials and the solid-phase bodies can have magnetic properties. The solid-phase bodies are used for isolating the microorganisms and for isolating the plasmid DNA.
Claims8
A method of isolating plasmid DNA from a microorganism culture with the aid of solid-phase bodies, comprising:
- a) acidifying said culture to an acid pH and mixing said culture with solid-phase bodies which are underivatized with antibody and which nonspecifically immobilize microorganisms thereon,
- b) resuspending the immobilized microorganisms, lysing the microorganisms, adding a neutralising binding buffer and incubating the buffer with the lysed microorganisms, followed by removing and discarding the solid-phase bodies,
- c) mixing the supernatant from step (b), containing plasmid DNA, with solid-phase bodies and incubating the supernatant with the bodies followed by eluting the plasmid DNA from the solid-phase bodies with an elution buffer.
A method of isolating plasmid DNA from a microorganism culture with the aid of magnetic solid-phase bodies having a solid gel surface, comprising:
- a) acidifying said culture to an acid pH and mixing said culture with magnetic solid-phase bodies having a solid gel surface which nonspecifically immobilize microoganisms thereon,
- b) resuspending the immobilized microorganisms, lysing the microorganisms, adding a neutralising binding buffer and incubating the buffer with the lysed microorganisms, followed by removing and discarding the magnetic solid-phase bodies having a solid gel surface,
- c) mixing the supernatant from step (b), containing plasmid DNA, with magnetic solid-phase bodies having a solid gel surface and incubating the supernatant with the bodies followed by eluting the plasmid DNA from the magnetic solid-phase bodies having a solid gel surface with an elution buffer.
Dependencies are inferred from the claim text.