JPH08166370A - Electrophoresis device and supporter manufacturing apparatus for electrophoresis device - Google Patents

Abstract

PURPOSE: To obtain the electrophoresis device, which can perform electrophoresis a for many kinds of materials in large amounts under the same conditions quickly, simply and inexpensively, i.e., at the industrial level also in the narrow space, and to obtain the supporter manufacturing apparatus, which can be also used as the electrophoresis device. CONSTITUTION: In a supporter box 7, a plurality of partitioning walls are contained, and an opening part 12 for electric conduction that can be sealed is provided at the bottom part. An upper part 1 in the supporter box can contain buffer liquid. In the inside of the box, a plurality of partitioning spaces, whose openings in the upper and lower directions are formed of a plurality of the partitioning walls, are formed. Furthermore, the apparatus comprises the supporter box 7, wherein a plurality of the partitioning spaces are mutually communicated at the bottom part, a supporter, which is filled in each partitioning space in the supporter box and filled with buffer liquid, and a lower buffer-liquid tank 2, which contains the supporter box 7, and wherein the opening part 12 for electric conduction and the buffer liquid are in contact. Furthermore, electrodes 3 and 5, which can apply the voltages in the up and down directions for the supporters, are interconnected in both upper and lower buffer liquids.

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an electrophoretic device,
More specifically, the present invention relates to an electrophoretic device capable of performing electrophoresis under the same conditions for various types of specimen samples, and an electrophoretic device that can also serve as a support (gel) manufacturing device. [0002] Electrophoresis is a core technology used in a very wide range of fields such as molecular biology and related fields, biophysics, clinical chemistry, and is used for proteins, sugars, DNA, RNA and the like. Used for separation, detection and purification. According to this method, when a sample solution is attached to one end of a support saturated with a buffer solution and an appropriate voltage is applied to both ends of the support, each component of the sample moves at a specific speed in the support. Is used. This moving speed depends on the molecular weight, charge amount, etc. of each component. In particular, gel electrophoresis in which the support is a gel includes agarose gel electrophoresis, polyacrylamide gel electrophoresis, gel disc electrophoresis, SDS gel electrophoresis, isoelectric focusing, and the like. It has been a driving force for the remarkable development of genetic engineering and protein engineering in recent years. In the production of such a support for an electrophoretic device, for example, the length is about 40 cm and the width is about 30.
cm glass plates facing each other through a spacer having a thickness of about 0.35 mm, and a support solution saturated with a buffer solution is filled between the glass plates so as to prevent bubbles and the like and solidified. After preparing the support, the support is loaded into an electrophoresis apparatus, and the upper and lower parts of the support are brought into contact with a buffer solution, respectively,
Form a well on the support and set the sample for analysis,
Electrophoresis is carried out by applying a voltage in the vertical direction of the support. [0005] When genetic engineering and protein engineering have come to be used in agriculture, pharmaceutical industry, animal and plant genome analysis, etc., the frequency of use of gel electrophoresis will increase dramatically. Although it has become necessary to rapidly analyze many types of specimen samples, the current condition is that supports for electrophoretic devices are prepared one by one during analysis, and care should be taken not to include bubbles and the like. In addition, the curing of the support becomes uneven depending on the manufacturing conditions, and since the two glass plates are usually fixed with a clip or the like, the liquid may not flow between the glass plate and the spacer. There is a problem of leakage, which is a complicated task. Moreover,
In recent years, it has been required to process a large amount of specimen samples under the same conditions at the same time, and it is desired that a homogeneous and large number of supports can be simultaneously prepared and that electrophoresis can be easily performed. ing. The first object of the present invention is to solve the above-mentioned problems and to perform electrophoresis of a large amount of various samples under the same conditions rapidly, simply and inexpensively, that is, on an industrial level in a narrow space. Is provided. A second object of the present invention is to prepare a support used for an electrophoresis apparatus capable of performing electrophoresis on a large amount of various kinds of samples under the same conditions rapidly, simply and inexpensively, that is, on an industrial level and in a narrow space. An object of the present invention is to provide a device for producing a support, which can also be used as an electrophoretic device. The electrophoretic device according to the present invention is a support box in which a plurality of partition walls are housed and an opening for energization is provided at the bottom. The upper part of is able to store the buffer solution,
A plurality of partition spaces whose interior is opened in the vertical direction is formed by a plurality of partition walls, and further, a support box in which the plurality of partition spaces communicate with each other at the bottom, and a partition space in the support box. It consists of a support filled inside and saturated with a buffer solution, and a lower buffer solution tank for accommodating the support box and bringing an opening for energization into contact with the buffer solution. It is characterized in that electrodes for applying a voltage to the support in the vertical direction are respectively wired. The electrophoretic device support production apparatus of the present invention is a support box in which a plurality of partition walls are housed and a bottom portion is provided with a sealable energization opening. Inside the body box, a plurality of partition walls are formed with a plurality of partition walls to open in the vertical direction.
In a support box in which the plurality of partition spaces are in communication with each other at the bottom, after the energizing opening is sealed, a support saturated with a buffer solution from the bottom is provided in the partition space in the support box. The present invention is characterized in that a body solution is filled and solidified to form a support, and then an opening for energization is opened. According to the present invention, by dividing the space inside the electrophoretic device with a glass plate or the like, a required number of supports can be simultaneously prepared, and the prepared supports can be used in the electrophoretic device. The sample can be directly attached to a part of the support without performing movement for incorporation or changing the shape of the prepared support, and electrophoresis can be performed. It is possible to perform the operations from the preparation of the sample to the electrophoresis in the same device, and it is possible to perform the electrophoresis of many kinds of samples under the same conditions. Embodiments of the present invention will be described below with reference to the drawings, but the present invention is not limited thereto. FIG. 1 is a perspective view of the electrophoretic device of the present invention.
FIG. 2 is a diagram for explaining a method of accommodating the partition wall in the support box, and FIG. 3 is a diagram for explaining a partition space portion in a sectional view when the electrophoresis device is used. In the figure,
1, 2 are tanks for buffer solution, 3 and 5 are electrodes (platinum wire) wired at the bottom of the buffer solution tank, 4 and 6 are anode and cathode connectors, 7 is a support box, and 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 energizing, 13 is a bolt, 14 is a groove for injecting a support solution, 15 is a sample, 16 is a sample fixing gel, 17 Indicates the support (gel saturated with buffer). An apparatus for preparing a support in the electrophoresis apparatus of the present invention will be described. The support box 7 is, for example, as shown in FIG. 2, a rectangular parallelepiped box body made of an insulating material such as plastic, and has a plurality of partition walls 8 inside with the top and one side face opened. Are stacked and stored via a spacer 9 (plastic film having a film thickness of 1 mm) to form a partition space. Since the plurality of partition walls 8 are housed on the stepped portions provided at both lower corners in the support box, the plurality of partition spaces are in communication with each other at the bottom portion. Next, the opening in the side surface portion is covered with the lid 11, and the plurality of partition walls 8 are fixed in the support box by using the bolt 13. The partition wall 8 is made of a glass plate, an acrylic plate or the like, and has a support (gel) on at least one surface.
Sigma coat (SIGM
It is preferable that the peeling treatment is performed by A) or the like. Further, if the number of partition walls 8 is 3, there are two partition spaces, and
Although individual supports can be produced at the same time, in the electrophoresis device of the present invention, the number of partition walls can be three or more,
For example, it has been confirmed that it can be 17 sheets. 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 film thickness of the spacer 9 can be various according to the purpose. A current-carrying opening 12 is opened in the lower portion of the lid 11, and is sealed with an adhesive tape or the like when the support is manufactured. As shown in the drawing, the upper portion of the support box 7 is widened and opened for the wiring of the electrodes 3.
The height of the partition wall 8 is set to be lower than the upper end of the support box 7. A thin tube such as a pipette is used inside the side wall of the support box 7 so that the support solution can be injected from the top of the support box 7 from the bottom of the support box 7. A groove 14 is provided. After assembling the support manufacturing apparatus in this way, a thin tube is inserted to the bottom of the support box through the groove 14 for injecting the support solution, and the support solution is lifted from the lower part to the upper part of the space. While injecting, inject through a thin tube. By injecting the support solution from the bottom of the support box, the support solution can be filled in the plurality of partition spaces without bubbles and the like. As the support solution, in the case of agarose gel electrophoresis, an ordinary 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 tris buffer saturated aqueous solution containing acrylamide, bisacrylamide, an ammonium persulfate catalyst, a polymerization initiator (N, N, N ', N'-tetramethylethylenediamine), etc. and maintained at a low temperature of about 4 ° C. Can be used. The support solution was injected to a height of about 1 cm below the upper end of the partition wall as in the case of an ordinary electrophoresis apparatus, and then a comb for forming wells was inserted and left at room temperature.
Allow the gel to solidify. After the support is prepared in this way, the adhesive tape sealing the current-carrying opening 12 is peeled off, and then a buffer solution having a 1-fold concentration is placed in the lower buffer tank 2 shown in FIG. Insert up to the height of the opening 12 for energization. Then, the comb is taken out, and as shown in FIG. 3, for example, a DNA sample is placed in the well formed by the comb, the gel solution for sample fixation is gelled and fixed, and the buffer solution is stored in the buffer tank 1. It is better to put it in the electrode and make it conductive with the electrode 3. In order to carry out electrophoresis, it is advisable to connect the anode and cathode connectors to a power source, apply a voltage of, for example, 100 V to 150 V between the electrodes 3 and 5, and perform electrophoresis for 20 hours to 24 hours. As described above, in the present invention, the apparatus for preparing a support for an electrophoretic device can be immediately used as an electrophoretic device. In the electrophoretic device of the present invention,
Like the usual electrophoretic device, a temperature control device such as a cooling device may be attached to the support box. After the electrophoresis, the lid 1 of the support box is
Except for 1, remove the glass plate one by one, take out the gel adhering to the glass plate, analyze it,
Alternatively, the specific component is extracted from the gel.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a perspective view of an electrophoretic device 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 it is used as an electrophoretic device.

Claims (1)

Claim: What is claimed is: 1. A support box in which a plurality of partition walls are stored and an opening for energization is provided in a bottom portion, and an upper portion of the support box can store a buffer solution. A plurality of partition walls are formed in the interior to form a plurality of partition spaces open in the vertical direction, and further, the plurality of partition spaces are in communication with each other at their bottoms, and the support box. It is composed of a support body filled in the partition space inside and saturated with a buffer solution, and a lower buffer solution tank for accommodating the support box and bringing the opening for energization into contact with the buffer solution. An electrophoretic device in which electrodes for applying a voltage to a support in a vertical direction are respectively wired in a buffer solution. 2. The electrophoresis device according to claim 1, wherein the support is an agarose gel or a polyacrylamide gel. 1. A support box which accommodates a plurality of partition walls and is provided with a sealable opening for energization at the bottom thereof, wherein the support box has a plurality of partition walls extending in the vertical direction. In a support box in which a plurality of open partition spaces are formed and the plurality of partition spaces are in communication with each other at the bottom, after partitioning the energizing opening, the partition space in the support box A support preparation device for an electrophoretic device, comprising: filling a support solution saturated with a buffer solution from the bottom thereof, solidifying the support solution to form a support, and then opening an opening for energization.