Liquid purification or separation – Processes – Liquid/liquid solvent or colloidal extraction or diffusing...
Patent
1997-12-08
1999-11-23
Weber, Jon P.
Liquid purification or separation
Processes
Liquid/liquid solvent or colloidal extraction or diffusing...
2103216, 2103231, 21050021, 210378, 422101, 422239, 435400, B01D 6302, B01D 1104, C12N 502
Patent
active
059894318
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a method of, and apparatus for, extraction of DNA from a suspension of cells. In particular, the invention relates to a method of and apparatus for, the extraction of plasmid DNA from micro-organisms and genomic DNA from micro-organisms and animal cells.
BACKGROUND ART
The application of recombinant DNA technology frequently involves the use of plasmids or cosmids in the cloning or manipulation of the target DNA. Typically, plasmids or cosmids are amplified by culturing micro-organisms harbouring plasmid or cosmid and ultimately extracting the plasmid or cosmid DNA from the culture. Purification of the extracted DNA is often required.
While automated or semi-automated procedures have been developed for some recombinant DNA techniques, particularly DNA purification and sequencing, many methods are still carried out manually. This is particularly the case for plasmid DNA extraction methods. Conventional methods, such as lysis by alkali or lysis by boiling (see Sambook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989) comprise a number of steps, the success of which rely on the skill of the worker. Repetitive application of such methods can thus lead to variation in the outcome of the procedures in the hands of an inexperienced worker.
It is often the case that plasmid or cosmid DNA must be extracted from a number of different cultures. Since micro-titre plates are used in some recombinant DNA techniques, it can be the case that extraction of DNA from hundreds of different cultures is desired. Conventional extraction procedures are not amenable to automation, or even semi-automation. Consequently, extraction of plasmid or cosmid DNA from a large number of individual cultures can constitute a time consuming and labour intensive operation.
There is therefore a need for a method which can be used to extract plasmid or cosmid DNA from cultures of micro-organisms which is simpler and more reproducible than existing procedures. There is also a need for an extraction method which is amenable to automation so that large numbers of cultures can be processed.
Similarly, the application of recombinant DNA technology and DNA analysis involves the use of genomic DNA from animal blood and other body fluids. Conventional methods such as extraction with organic solvents (see Sambook et al., Molecular Cloning: A Laboratory Manual, 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989) comprise a number of steps, the success of which rely on the skill of the worker. Repetitive application of such methods can thus lead to variation in the outcome of the procedures in the hands of an inexperienced worker.
It is often the case that genomic DNA must be extracted from large numbers of samples. This is particularly the case for use of DNA in the polymerase chain reaction and DNA sequencing. Since microtitre plates are used in some recombinant DNA and genetic analysis techniques, it can be the case that DNA from hundreds of samples is required.
There is therefore a need for a method which can be used to extract genomic DNA from blood and other body fluids which is simpler and more reproducible than existing procedures. There is also a need for an extraction method which is amenable to automation so that large numbers of samples can be processed.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide a method of extracting DNA from a suspension of cells which might overcome the disadvantages of existing extraction methods. In particular, it is an object of the invention to provide a method of selectively extracting plasmid DNA from a suspension of cells which might overcome the disadvantages of existing methods for the extraction of plasmid DNA.
It is a further object of the invention to provide apparatus for use in extracting DNA from a suspension of cells.
The term "suspension of cells" as used above and hereafter includes a culture of animal cells, a culture of mic
REFERENCES:
Hirasaki et al., Anim. Cell Technol.: Dev. 21st Century, [Proc. Meet.], Meeting Date 1994, pages 547-551. Editor(s): Beuvery, E. Coen; Griffiths, J. Brian; Zeijlemaker, Wim P. Publisher: Kluwer, Dordrecht, Neth., in Chem Abstr., vol. 125, Abstr. No. 563, 1995.
Gupta, et al.; Hemaphereals; International Journal of Artificial Organs, vol. 14, No. 1, (1990) "Comparison of polyamide and polypropylene membranes for plasma separation", pp. 56-60.
Evans Timothy Martin
Hugh Robert Don
Progen Industries Ltd
Weber Jon P.
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