JP3541971B2 - Analysis method using electrophoresis device - Google Patents

Description

[0001]
[Industrial applications]
The present invention relates to an electrophoresis apparatus, and more particularly, to an electrophoresis apparatus capable of performing electrophoresis on a variety of sample samples under the same conditions, and an electrophoresis apparatus which can also be used as an apparatus for producing a support (gel).
[0002]
[Prior art]
Electrophoresis is a fundamental technology used in a very wide range of fields such as molecular biology and related fields, biophysics and clinical chemistry, and is used for the separation, detection and purification of proteins, sugars, DNA, RNA, etc. ing.
[0003]
This method utilizes a method in which a sample solution is attached to one end of a support saturated with a buffer solution, and when an appropriate voltage is applied to both ends, each component of the sample moves at a specific speed in the support. Things. This moving speed depends on the molecular weight, charge amount, and the like of each component. In particular, the gel electrophoresis in which the support is a gel includes agarose gel electrophoresis, polyacrylamide gel electrophoresis, gel disk electrophoresis, SDS gel electrophoresis, isoelectric focusing, and the like. In recent years, it has been the driving force for the remarkable development of genetic engineering and protein engineering.
[0004]
In producing such a support in an electrophoresis apparatus, for example, two glass plates having a length of about 40 cm and a width of about 30 cm are opposed to each other with a spacer having a thickness of about 0.35 mm interposed therebetween. A support solution saturated with a buffer solution is filled so as not to contain air bubbles and the like, and solidified to prepare a support. The support is then loaded into an electrophoresis apparatus, and the upper and lower portions of the support are brought into contact with the buffer solution, respectively. At the same time, a well is formed on the support, an analysis sample is set, and a voltage is applied in the vertical direction of the support to perform electrophoresis.
[0005]
[Problems to be solved by the invention]
When genetic engineering and protein engineering are used in agriculture, the pharmaceutical industry, etc. or genome analysis of animals and plants, the frequency of gel electrophoresis will dramatically increase, and it will be necessary to analyze many types of sample samples quickly. At present, the support for the electrophoresis apparatus is prepared one by one at the time of analysis, and it needs to be carefully prepared so as not to contain bubbles, etc. In some cases, the curing of the support is not uniform, and furthermore, since the two glass plates are usually fixed with clips or the like, there is a problem that the liquid leaks from between the glass plate and the spacer, which is a troublesome work. Has become. Moreover, in recent years, it has been required to simultaneously process a large number of sample samples under the same conditions, and it is necessary to simultaneously prepare a large number of supports that are homogeneous and to easily perform electrophoresis. Is desired.
[0006]
A first object of the present invention is to solve the above problems and provide an electrophoresis apparatus capable of electrophoresing a large amount of various samples under the same conditions, quickly, simply and inexpensively, that is, at an industrial level and in a narrow space. is there.
[0007]
A second object of the present invention is an apparatus for preparing a support for use in an electrophoresis apparatus capable of electrophoresing a large amount of various samples under the same conditions, quickly, simply, and inexpensively, that is, in an industrial level and in a narrow space. Another object of the present invention is to provide an apparatus for producing a support that can also be used as an electrophoresis apparatus.
[0008]
[Means for Solving the Problems]
An analysis method using the electrophoresis apparatus of the present invention is a support box in which a plurality of partition walls subjected to a peeling treatment are stored and a cover is provided with a current-carrying opening, and an upper portion of the support box is provided. A support body capable of accommodating a buffer solution, a plurality of partition spaces having the inside opened in the vertical direction by a plurality of partition walls, and the plurality of partition spaces being in communication with each other at the bottom. A box, a support filled with a partition space in the support box, an analysis sample is fixed at an upper end and saturated with a buffer, and the support box is housed. And a lower buffer tank to be brought into contact with the lower electrode, and further, electrodes for enabling a voltage to be applied vertically to the support in both upper and lower buffers are wired to form an electrophoresis apparatus, and both electrodes in the electrophoresis apparatus are used. After applying a voltage to the electrophoresis of the analysis sample, removing the lid in the support box, peeling off the partition wall, extracting the support attached to the partition wall, and extracting a specific component. It is characterized by.
[0010]
[Action and effect of the invention]
According to the present invention, a required number of supports can be simultaneously prepared by dividing the space in the electrophoresis apparatus with a glass plate or the like, and the prepared support is moved or manufactured for incorporation into the electrophoresis apparatus. This allows electrophoresis to be performed by attaching a sample to a part of the support without changing the shape of the support.The same method is used for the simple preparation of the support and the operation from preparation of the support to electrophoresis. This can be performed in an apparatus, and enables electrophoresis of various types of samples under the same conditions.
[0011]
Hereinafter, embodiments of the present invention will be described with reference to the drawings, but the present invention is not limited thereto.
[0012]
FIG. 1 is a perspective view of an electrophoresis apparatus of the present invention, FIG. 2 is a view for explaining a method of storing a partition wall in a support box, and FIG. 3 is a partition space portion when the electrophoresis apparatus is used. FIG. 4 is a view for explaining a section view. In the figure, reference numerals 1 and 2 denote buffer solution tanks, reference numerals 3 and 5 denote electrodes (platinum wires) respectively wired at the bottom of the buffer solution tank, reference numerals 4 and 6 denote anode and cathode connectors, reference numeral 7 denotes a support box, and reference numeral 8 Is a partition wall, 9 is a spacer, 10 is a packing, 11 is a lid of a support box, 12 is an opening for energization, 13 is a bolt, 14 is a groove for injecting a support solution, 15 is a sample, and 16 is a sample fixation. Gel, 17 indicates support (gel saturated with buffer).
[0013]
The support manufacturing apparatus in the electrophoresis apparatus of the present invention will be described. As shown in FIG. 2, for example, the support box 7 is a rectangular parallelepiped box formed of an insulating material such as plastic, and has a plurality of partition walls 8 inside thereof with the upper part and one side part opened. Are stacked one on top of the other via a spacer 9 (plastic film having a thickness of 1 mm) to form a partition space. The plurality of partition walls 8 are housed on steps provided at both lower corners in the support box, so that the plurality of partition spaces are in communication at the bottom. Then, the lid 11 is put on the opening in the side surface, and the plurality of partition walls 8 are fixed in the support box using the bolts 13.
[0014]
A glass plate, an acrylic plate, or the like is used for the partition wall 8, and at least one surface is subjected to a release treatment using a sigma coat (manufactured by SIGMA) or the like in order to improve the releasability from the support (gel). Good. Further, if the number of the partition walls 8 is three, the partition space becomes two places and two supports can be simultaneously manufactured. However, in the electrophoresis apparatus of the present invention, the number of the partition walls can be three or more. We have confirmed that we can have 17 images. Further, as shown in FIG. 3, the upper end of the partition wall may be formed in a tapered shape in order to easily store the analysis sample 15. The thickness of the spacer 9 can be various thicknesses depending on the purpose.
[0015]
An opening 12 for energization is opened in the lower part of the lid 11, and is sealed with an adhesive tape or the like when the support is manufactured.
[0016]
The upper part of the support box 7 is widened and opened for wiring of the electrodes 3 as shown in the figure, and the height of the partition wall 8 is set to be lower than the upper end of the support box 7. You.
[0017]
A groove 14 for injecting a support solution is provided inside the side wall of the support box 7 using a thin tube such as a pipette so that the support solution can be injected from the bottom of the support box 7 from the top of the support box 7. Provided.
[0018]
After assembling the support preparation apparatus in this manner, a thin tube is inserted through the support solution injection groove 14 to the bottom of the support box, and the support solution is partitioned while the partition space is raised from the lower part to the upper part. Inject through tubule. By injecting the support solution from the bottom of the support box, the support solution can be filled into a plurality of partition spaces without bubbles or the like. As the support solution, in the case of agarose gel electrophoresis, a usual support forming solution, for example, an aqueous solution of agarose (agar) saturated with Tris buffer and heated can be used. In the case of the method, a saturated aqueous solution of a tris buffer solution comprising acrylamide, bisacrylamide, an ammonium persulfate catalyst, a polymerization initiator (N, N, N ', N'-tetramethylethylenediamine), etc., maintained at a low temperature of about 4 ° C. Can be used.
[0019]
The support solution is poured to a height of about 1 cm below the upper end of the partition wall, as in a usual electrophoresis apparatus, and then a comb for forming a well is inserted and left at room temperature to solidify the gel.
[0020]
After preparing the support in this manner, the adhesive tape sealing the current-carrying opening 12 was peeled off, and then a buffer solution of 1-fold concentration was placed in the lower buffer solution tank 2 shown in FIG. Insert up to the height of the part 12.
[0021]
Next, the comb is taken out and, as shown in FIG. 3, for example, a DNA sample is placed in a well formed by the comb, the gel solution for fixing the sample is gelled and fixed, and the buffer solution is put into the buffer tank 1. It is good to conduct with the electrode 3.
[0022]
In order to perform electrophoresis, the anode and the cathode connectors may be connected to power supplies, and a voltage of, for example, 100 V to 150 V may be applied between the electrodes 3 and 5 to perform electrophoresis for 20 hours to 24 hours.
[0023]
As described above, in the present invention, the electrophoresis apparatus support producing apparatus can be immediately used as the electrophoresis apparatus.
[0024]
Further, in the electrophoresis apparatus of the present invention, a temperature control device such as a cooling device may be attached to the support box as in a normal electrophoresis apparatus.
[0025]
After completion of the electrophoresis, the glass plate is peeled off one by one except for the lid 11 of the support box, the gel attached to the glass plate is taken out, and the gel is analyzed or a specific component is extracted from the gel.
[Brief description of the drawings]
FIG. 1 is a perspective view of an electrophoresis apparatus of the present invention.
FIG. 2 is a diagram for explaining a method of storing a partition wall in a support box.
FIG. 3 is a diagram for explaining a partition space portion in a cross-sectional view when the electrophoresis device is used.

Claims (2)

A plurality of partition walls subjected to the peeling process are stored, and a support box provided with a current-carrying opening in a lid portion, wherein the upper portion of the support box is capable of storing a buffer solution, and the inside is A plurality of partition spaces opened in the vertical direction are formed by the plurality of partition walls, and further, the plurality of partition spaces are in a support box that is in a state of being communicated with each other at the bottom, and in a partition space in the support box. A support that is filled and has an analysis sample fixed at the upper end and saturated with a buffer solution, and a lower buffer tank that houses the support box and makes an opening for energization contact with the buffer solution. the electrophoresis apparatus support in a buffer of the upper and lower both in the vertical direction in the electrodes a voltage can be applied to wire respectively, said analysis sample is electrophoresed by applying a voltage between the electrodes in the electrophoretic device Was followed, except the lid in the support box, peeled off the partition wall, the analysis method using an electrophoresis apparatus characterized by the extraction of a specific component is taken out of the support attached to the partition wall. The method according to claim 1, wherein the support is an agarose gel or a polyacrylamide gel.

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