Chemistry: electrical and wave energy – Apparatus – Coating – forming or etching by sputtering
Patent
1990-04-30
1992-04-07
Niebling, John
Chemistry: electrical and wave energy
Apparatus
Coating, forming or etching by sputtering
2041801, 2041828, B01D 6142, B01D 5702, C25B 100
Patent
active
051025248
DESCRIPTION:
BRIEF SUMMARY
The invention relates to a multiple electrophoresis method and device ensuring the controlled migration of macromolecules in rectangular gel plates.
Techniques are used at the present time for separating macromolecules, such as proteins or nucleic acids, by electrophoresis using an electric field applied to the longitudinal ends of a rectangular plate of an appropriate gel, for example agarose or polyacrylamide. Samples of macromolecules, for example nucleic acids, to be separated are deposited in wells formed in the gel, along an edge of the plate then the whole is immersed in an appropriate electrophoresis liquid. The electrodes placed against said edge of the plate and the opposite edge thereof are connected to different potentials so that an electric field is created between the electrodes in a direction corresponding to the desired direction of migration of the macromolecules in the gel plate. Under the effect of the electric field, the macromolecules of the samples housed in the wells move towards the opposite edge of the plate, through the gel, at speeds which depend particularly on their molar mass so that, at the end of a given time, macromolecules of different molar masses have travelled through the gel over different distances.
In a known technique, the macromolecules thus separated are then transferred either by suction, or by means of an electric field oriented perpendicularly to the plate in the desired direction, on a membrane placed on a large face of the gel plate, for hybridization and subsequent detection thereof.
This technique and the apparatus which has been devised for implementing it relate essentially to the laboratory, i.e. it is a question of small-sized apparatus operating at a slow rate, processing the gel plates one after the other, etc.
Furthermore, the separations obtained with such apparatus are not perfectly repetitive and may vary from one plate to the other or from one sample to the other in the same plate, if only because the electric field developed between the two electrodes is not uniform, because of the heterogeneity of the medium passed through, so that identical macromolecules, having the same molar mass, may migrate over different distances if they are placed at different points on the same plate or on different plates.
The result is particularly difficulties in interpreting the results and the impossibility of automating the apparatus for macromolecule separation by electrophoresis.
The object of the invention is in particular to avoid such drawbacks of the prior technique.
It provides a multiple electrophoresis method and device for separating macromolecules in gel plates in a way which is reliable, faithful, repetitive and perfectly automatable.
Yet another object of the invention is a method and device of this type which make it possible to process simultaneously a large number of gel plates.
A further object of the invention is a method and device of the above type, making it possible to modify at will the conditions of separation of the macromolecules, particularly by controlled variation of the electric field applied to the gel plates for migration of the macromolecules.
For this, the invention provides a multiple electrophoresis method ensuring the controlled migration of macromolecules in rectangular gel plates, characterized in that it consists: them spaced apart so as to form at least a parallelepipedic stack, stack a plurality of elongate electrodes, for example in the form of wires, parallel to each other and to the planes of the plates, and contained in a series of planes perpendicular to a common desired migration direction of the macromolecules in the plates, electrophoresis liquid, migration direction to the same potential, and different planes so as to create in the bath and in the plates an electric field whose direction is at all points substantially parallel to the desired migration direction.
The electrodes contained in planes perpendicular to the desired migration direction of the macromolecules define equipotential surfaces which are subst
REFERENCES:
patent: 4088561 (1978-05-01), Anderson
patent: 4655898 (1987-04-01), Poulhes et al.
patent: 4994166 (1991-02-01), Fernwood et al.
Bertin & Cie
Koestner Caroline
Niebling John
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