Recovery of plasmids in an aqueous two-phase system

Details Recovery of plasmids in an aqueous two-phase system Recovery of plasmids in an aqueous two-phase system

2003-06-17

2009-06-30

Burkhart, Michael (Department: 1633)

Chemistry: molecular biology and microbiology

Process of utilizing an enzyme or micro-organism to destroy...

Treating animal or plant material or micro-organism

C435S091400, C435S320100

Reexamination Certificate

active

07553658

ABSTRACT:
The present invention is a method for the purification of plasmid DNA comprising to provide a composition comprising a first polymer having inverse solubility characteristics and a second polymer immiscible in the first polymer; contacting said solution with an aqueous solution comprising plasmid DNA; providing phase separation and isolating the aqueous phase; and increasing the temperature of the isolated phase to a temperature above the cloud point of the first polymer and below the temperature where plasmid DNA is degraded and subsequently isolating the aqueous phase so formed. The invention also encompasses a kit for purification of plasmid DNA as described above.

REFERENCES:
patent: 6107467 (2000-08-01), Ageland et al.
patent: W098/11140 (1998-03-01), None
Ohlsson R, (A rapid method for the isolation of circular DNA using an aqueous two-phase partition system. Nucleic Acids Res. Feb. 1978;5(2):583-90).
Persson, J., et al., “Purification of protein and recycling of polymers in a new aqueous two-phase system using two thermoseparating polymers”,Journal of Chromatography A, vol. 864, 1999, p. 31-48.
Tjerneld, F., et al., “Utilization of Temperature-induced Phase Separation for Purification of Biomolecules”,International Symposium on Separations for Biotechnology Reading: Separation for Biotechnology 3. Cambridge: Royal Society of Chemistry, 1994, p. 505-511.
Ribeiro, S., et al., “Isolation of Plasmid DNA from Cell Lysates by Aqueous Two-Phase Systems”,Biotechnology and Bioengineering, vol. 78, No. 4, May 2002, p. 376-384.

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