JPH0734365U - Horizontal gel electrophoresis device - Google Patents

Abstract

(57) [Summary] [Purpose] To simplify the liquid contact mechanism between the flat plate gel placed in the horizontal plane and the electrode solution, facilitate sample loading, and allow smooth migration. [Structure] Vertically rising rising parts 4 and 6 are formed at both ends of a flat gel gel 2 in which a migration part is installed in a horizontal plane. The compatible rising parts 4 and 6 are a cathode side electrode tank 8 and an anode side. It is in contact with each electrolytic solution in the electrode tank 10. At the end of the rising portion 4 on the cathode side, there is provided a sample introduction portion 12 formed by inserting a comb in the vertical direction. Electrode wires 14 and 16 are respectively immersed in the electrolytic solutions of both electrode tanks 8 and 10, and a migration voltage is applied from a high voltage power supply 18 between both electrode wires 14 and 16. In order to keep the temperature of the gel 2 uniform, a cooling water circulation mechanism 20 is provided on the lower surface side of the gel 2, and the cooling water is circulated by the cooling water control system 20 and kept at a constant temperature.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 The present invention relates to an electrophoretic device used for separating and analyzing proteins and determining the electric base sequence of DNA.

[0002]

[Prior art]

 Some gel electrophoresis devices use a flat gel (slab gel). There are two types of gel electrophoresis apparatus using a flat gel: one in which the migration surface of the flat gel is placed in a vertical plane to allow the sample to migrate, and another in which the migration surface is placed in the horizontal plane to migrate the sample.

When a sample is electrophoresed in a gel, an electrophoretic voltage is applied across the gel to cause a current to flow. The heat generated by the current causes a temperature distribution in the migration surface of the gel. Due to the temperature distribution, the migration speed becomes non-uniform in the plane of the gel, and a phenomenon called smileing occurs in the migrated pattern, which causes an error in the DNA sequencing.

A horizontal gel electrophoresis apparatus in which the migration surface of a flat gel is set in a horizontal plane has an excellent effect of radiating heat from the gel, and it is easy to uniformly cool the gel to reduce the temperature distribution. Has advantages. In a horizontal gel electrophoresis device, a comb is inserted at one end of a flat gel placed in a horizontal plane in a direction perpendicular to the surface of the gel to form a slot for sample injection (eg, “Nucleic Acids Research”). Vol. 19, No. 15, pp. 4121-4126). The slab gel used there exists only in the horizontal plane. Therefore, on the cathode side, to insert a comb to form a slot for sample injection and to make contact with the cathode side electrolyte solution, and on the anode side to contact the anode side electrolyte solution. , There are holes at both ends of the upper gel plate that holds the gel from above.

[0005]

[Problems to be solved by the device]

 In the horizontal gel electrophoresis device introduced in the above-mentioned literature, it is necessary to achieve the liquid contact between the flat plate gel in the horizontal direction and the electrolytic solution in the electrode tank through the hole formed in the upper gel plate. There is a problem that becomes complicated. In addition, the comb forming the slot for the sample must be inserted vertically from the gel in the horizontal direction. When the thickness of the gel is as thin as 1 mm or less, it is not easy to insert the comb to the correct depth from the vertical direction on the surface of such a thin gel, and the tip of the comb is placed in the thickness direction of the gel. It may break through. In that case, it becomes difficult to smoothly migrate the sample. An object of the present invention is to simplify a liquid contact mechanism between a flat gel whose electrophoretic section is placed in a horizontal plane and an electrode liquid, facilitate sample loading, and allow smooth migration.

[0006]

[Means for Solving the Problems]

 The electrophoretic device of the present invention is provided such that the electrophoretic portion is installed in a horizontal plane, and the flat gel having vertical rising portions at both ends thereof and the gel rising portions at both ends of the flat gel are respectively in contact with the electrolytic solution. It is a horizontal gel electrophoresis apparatus provided with an electrode bath for a cathode and an anode, and an electrophoretic power supply device for applying an electrophoretic voltage between the electrode baths.

[0007]

[Action]

 In the present invention, since both ends of the flat gel in which the electrophoretic part is installed in the horizontal plane are raised in the vertical direction, the liquid contact mechanism with the electrolytic solution is simple. Since the comb into which the sample is loaded can be inserted into the end surface of the gel which has been vertically raised, the comb can be easily inserted, and the sample is correctly loaded into the gel and migration is smoothly performed.

[0008]

【Example】

 FIG. 1 schematically shows the present invention. Standing portions 4 and 6 that stand in the vertical direction are formed at both ends of the flat gel 2 in which the electrophoretic portion is installed in the horizontal plane. The flat gel 2 is a polyacrylamide gel, and is formed in a state of being sandwiched between two glass plates together with the rising portions 4 and 6 at both ends thereof. The compatibility parts 4 and 6 are in contact with the respective electrolytic solutions of the cathode side electrode tank 8 and the anode side electrode tank 10.

At the end of the rising portion 4 on the cathode side, a comb is inserted in the vertical direction, and a sample introduction portion 12 formed after the comb is removed is provided. A protein sample or a DNA fragment sample is input to the sample input unit 12. In the case of a DNA fragment sample, for example, samples of terminal base types A (adenine), G (guanine), T (thymine), and C (cytosine) that have been pretreated by the Sanger's method are prepared. The cells are put into different slots of the feeding section 12 and are allowed to swim in different migration lanes at the same time.

Electrode wires 14 and 16 are respectively immersed in the electrolytic solutions of both electrode tanks 8 and 10, and a migration voltage is applied from a high voltage power source 18 between both electrode wires 14 and 16. In order to keep the temperature of the gel 2 uniform, a cooling water circulation mechanism 20 is provided on the lower surface side of the gel 2, and the cooling water is circulated by the cooling water control system 20 to maintain a constant temperature.

2 to 4 show a specific structure of one embodiment. In order to form a gel, as shown in FIG. 3 (A), a flat lower gel plate 30 and a frame-shaped spacer 32 placed thereon, and a flat plate placed further thereon, A pair of gel plates 31 is constituted by an upper gel plate 34 having grooves 36 and 38 corresponding to positions inside the frame of the spacer 32 at both ends in the longitudinal direction. The three gel plates 30, 32, and 34 are combined and placed in the horizontal direction, and the spacer 32 is sandwiched between the lower gel plate 30 and the upper gel plate 36 in the horizontal space, and the both ends thereof rise vertically. The space defined by the grooves 36 and 38 is filled with gel. As a result, as shown in FIG. 3B, a tabular gel having the tabular gel 2 of the electrophoretic section and the rising portions 4 and 6 at both ends thereof is formed. For example, a 6% polyacrylamide gel is used as the gel, and the gel thickness determined by the thickness of the spacer 32 is, for example, 0. 25 mm.

As shown in FIG. 2, the cooling water circulation system 20 installed on the lower side of the lower gel plate 30 is supplied with cooling water from the cooling water control system 22 via the cooling water inflow part 21 a. The cooling water that has cooled the gel is returned to the cooling water control system 22 from the cooling water outlet 21b of the cooling water circulation mechanism 20. The cooling water is controlled to a constant temperature and circulates between the cooling water circulation mechanism 20 and the cooling water control system 22.

The electrode tanks 8 and 10 are placed on the grooves 36 and 38 at both ends of the upper gel plate 34 of the set of gel plates 31 placed on the cooling water circulation mechanism 20, respectively. Both electrode tanks 8 and 10 are detachably fixed to a cooling water circulation mechanism 20 by an electrode tank fixture 9.

The electrode tanks 8 and 10 have symmetrical shapes. Although one electrode tank 8 is shown in FIG. 4, the other electrode tank 10 has a similar structure. The electrode tanks 8 and 10 are open at the top and bottom, and the openings in the bottom allow the gel in the grooves 36 and 38 of the upper gel plate to come into contact with the electrolytic solution. A seal member 40 such as an O-ring or sponge is provided at the bottom of the electrode tanks 8 and 10 in order to prevent liquid leakage while being pressed by the upper gel plate 34.

An electrode wire 14 is provided in the cathode-side electrode tank 8 via a cathode voltage applying plug 15, and the electrode wire 14 extends into the electrolytic tank 8 and is immersed in an electrolytic solution. Similarly, in the anode-side electrode tank 10, the electrode wire 16 is provided through the anode voltage applying plug 17, and the electrode wire 16 extends into the electrolytic tank 10 and is immersed in the electrolytic solution. By removing the electrode bath fixture 9, the electrode baths 8 and 10 can be removed from the cooling water circulation mechanism 20, and the gel plate 31 can also be removed to take out the gel.

The operation of the embodiment shown in FIGS. 2 to 4 will be described. The gel plate 31 is assembled and filled with polyacrylamide gel, and a comb is inserted into the rising part of the gel formed in one groove 36 of the upper gel plate 34 to form a sample input part. A sample such as a DNA fragment sample is put in the sample input section. After that, both electrode tanks 8 and 10 are placed on the gel plate 31, the electrode tanks 8 and 10 are fixed to the cooling water circulation mechanism 20 by the fixture 9, and both electrode tanks 8 and 10 are filled with the electrolytic solution. After that, a high voltage power source 18 (FIG. 1) is connected between both plugs 15 and 17 to apply a migration voltage. At this time, cooling water is circulated in the cooling water circulation mechanism 20 from the cooling water control system 22 to keep the temperature of the gel constant.

When the sample introduced from the sample introduction section is a DNA fragment sample prepared by the Sanger method for each terminal salt group, the sample is labeled with a fluorescent substance. When the sample migrates through gel 2 and is separated, the application of the migration voltage is stopped, and the migration band developed in the gel is irradiated with excitation light and the fluorescence is detected to read. The labeling compound is not limited to the fluorescent substance and may be a radioactive substance. It may also be read by other known methods without labeling. If the upper gel plate 34 is made of a transparent glass plate, the gel can be read while being sandwiched in the gel plate 31.

[0018]

[Effect of device]

 In the present invention, since both ends of the flat plate gel in which the electrophoretic part is installed horizontally stand upright, the comb for loading the sample is inserted in the same vertical direction from the end of the vertical standup part of the gel. Therefore, it is easy to form the sample input part correctly, and migration can be smoothly caused. When the comb is inserted vertically from a horizontal gel as proposed in the literature, the comb must penetrate the gel to form a good sample input, especially if the gel is thin. However, such problems do not occur in the present invention. As described above, according to the present invention, the electrophoresis of the sample can be smoothly performed in the horizontal gel electrophoresis apparatus with improved thermal effect. Further, since both ends of the gel stand vertically from the flat surface of the gel, the liquid contact with the electrolytic solution in both electrode tanks is facilitated and the structure of the liquid contact part is simplified.

[Brief description of drawings]

FIG. 1 is a schematic configuration diagram showing the present invention.

FIG. 2 is a perspective view showing an example.

FIG. 3A is an exploded plan view of the gel plate in the same embodiment, and FIG.
It is sectional drawing which shows the state cut | disconnected in the X'direction.

FIG. 4 is a view showing an electrode tank in the same example,
(A) is a plan view and (B) is a sectional view taken along the line YY '.

[Explanation of symbols]

 2 Gel migration part 4,6 Gel rising part 8,10 Electrode tank 12 Sample input part 14,16 Electrode wire 18 High voltage power supply 20 Cooling water circulation mechanism

Front page continued (72) Shuji Fujibun, 1st Nishinokyo Kuwabara-cho, Nakagyo-ku, Kyoto City, Kyoto Prefecture Shimadzu Corporation Sanjo Factory (72) Inventor Yoshishi Togawa, Nishinokyo Kuwabara-cho, Nakagyo-ku, Kyoto City, Kyoto Prefecture Shimazu Corporation Factory Sanjo Factory

Claims (1)

[Scope of utility model registration request] 1. A flat gel having electrophoretic portions arranged in a horizontal plane and having vertical rising portions at both ends thereof, and a cathode provided so as to contact gel rising portions at both ends of the flat gel with an electrolytic solution, respectively. A horizontal gel electrophoresis apparatus comprising: an electrode bath for an anode and an electrophoretic power supply device for applying an electrophoretic voltage between the electrode baths.