Cartridge for preparing purified nucleic acids

Details Cartridge for preparing purified nucleic acids Cartridge for preparing purified nucleic acids

1996-05-01

1998-08-04

Beisner, William H.

Chemistry: molecular biology and microbiology

Apparatus

Involving lysis of a microorganism by means other than...

4352971, 435270, 422101, 21032164, C12M 112, C12M 306

Patent

active

057892431

DESCRIPTION:

BRIEF SUMMARY
FIELD AND BACKGROUND OF THE INVENTION

The invention relates to a cartridge for extracting and purifying nucleic acids, in particular genome DNA, from a blood sample, a tissue cell sample, or a culture cell sample.
International application WO 93/01312 has already disclosed a cartridge of this type constituting a considerable advance over competing devices by virtue of its ease of use, its efficiency, its reliability, and the economic conditions required for manufacturing and using it.
The cartridge described in that prior application comprises a flat frame having a dialysis membrane stuck to each of the two faces thereof, thereby defining a dialysis enclosure between them. A plane filter for retaining cell nuclei is mounted inside the frame between the membranes, and means are provided for causing the following to flow between the membranes: a sample of lysed cells; substances for washing and rinsing the nuclei retained by the filter; and substances for lysing the nuclei and degrading proteins. The nucleic acids released into the enclosure are purified by dialysis. For some cells, the operations of cell lysis, nucleus lysis, degradation of proteins and other contaminants, and dialysis can all be done directly in the enclosure.
The cartridge serves to automate the operations of extracting and purifying nucleic acids, which operations take place in the cartridge without it being manipulated.


SUMMARY OF THE INVENTION

The present invention seeks to improve that cartridge, aiming to simplify its structure and its manufacture, to reduce its cost, and to improve its operation, its performance, and its reliability.
To this end, the invention provides a cartridge for preparing purified nucleic acids, the cartridge comprising a dialysis enclosure, insertion means for inserting a sample of cells and reaction substances into said enclosure, discharge means for discharging reaction substances and purified nucleic acids, and a filter for retaining cell nuclei, the filter being disposed inside the dialysis enclosure, the cartridge being characterized in that the filter and the dialysis enclosure are constituted by a flexible filter tube and a flexible dialysis tube placed one within the other, the filter tube being open at one end and closed at the other, the dialysis tube being open at both ends, said ends being mounted together with at least the open end of the filter tube in endpieces of plastics material or of elastomer designed to receive the above-mentioned insertion and discharge means.
The flexible tubular structures of the filter and of the dialysis enclosure enable the system to be drained completely by suction, where the effect of the suction is to flatten the filter tube and the dialysis tube onto themselves so as to leave no residual internal volume. In addition, the dialysis enclosure contains no foreign body other than the filter tube, thereby reducing the risk of contamination or of pollution.
In a preferred embodiment of the invention, each endpiece includes a hollow body or cap of material that is self-sealing after being pierced, receiving an open end of the filter tube and/or of the dialysis tube, and embedded in a sealant.
The endpieces are also rigidly interconnected, e.g. by longitudinal uprights which are fixed by mutual engagement and/or adhesive to the endpieces.
In general, the cartridge of the invention is designed to be placed in a tube or a housing connected to means for circulating a dialysis liquid.


BRIEF DESCRIPTION OF THE DRAWINGS

The invention will be better understood and other characteristics, details, and advantages thereof will appear more clearly on reading the following description given by way of example and made with reference to the accompanying drawings, in which:
FIG. 1 is a diagram of a cartridge of the invention;
FIG. 2 is a diagram of a tube designed to receive the cartridge;
FIGS. 3 and 4 are diagrams of the filter tube and of the dialysis tube;
FIG. 5 is a diagram showing a step during the mounting of a cartridge of the invention;
FIG. 6 is a perspective diagram o

REFERENCES:
patent: 4787963 (1988-11-01), MacConnell
patent: 4921952 (1990-05-01), Longmire et al.
patent: 5352609 (1994-10-01), Boquet

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