JP4712364B2 - Electrophoresis device electrophoresis tank - Google Patents

Description

The present invention mainly uses biomolecules such as DNA, RNA, and proteins, or mixtures of various low-molecular compounds and complexes having a charge, and electrophoresis used for separation analysis, purification, and recovery of these samples. related to the electrophoresis tank of instrumentation 置用.

As a conventional electrophoresis apparatus, an electrophoresis apparatus disclosed in Japanese Utility Model Publication No. 63-39639 is known. This electrophoresis apparatus is composed only of a fuse, a rectifier, an operation on / off switch, and an output changeover switch, and can supply a current corresponding to the impedance of the electrophoresis carrier.
Shoko Sho 63-39639 All of the electrophoresis methods (Medical and Dental Publishing Co., Ltd., 1983) In all of the electrophoresis methods (Medical and Dental Publishing Co., Ltd., 1983), electrodes are placed so as to sandwich the gel soaked in the buffer solution. In addition, a plate-like body having partitioning properties protruding from the buffer solution is disposed between the electrode and the gel, and a configuration in which the plate-like body and the bottom of the electrophoresis tank are used as an electrical passage is shown. .

However, since the inventive device is directly connected to a general-purpose AC power source such as an AC 100 V household power source, there is a high possibility that a large amount of current flows due to a rapid drop in impedance of the electrophoresis carrier or a short circuit of each part. Although this large amount of current is dangerous for the human body, the only consideration for electrical insulation is the disconnection of the built-in fuse and the protective functional form of the case housing the electronic circuit.
Such a simple protection function is not enough at present when electrophoresis experiments are increasing as part of education at high schools. In addition, if an unhandled person conducts an experiment, there is a high possibility that the device will fail, such as accidentally spilling liquid, so that it is difficult to make a configuration that does not cause failure or operations that may cause failure. Such an improvement in the feeling of use is required.

However, if the circuit configuration is complicated only to improve safety and usability, or if automation is promoted, the size and cost of the apparatus will increase. As a result, such devices can only be handled by specialized research institutes, and do not lead to expansion of educational applications or speeding up research by handling multiple units by a single researcher. Therefore, there is a need for an electrophoresis apparatus that is as simple as possible and safe and easy to operate.
The inventors have devised a simple and safe electrophoresis apparatus in Japanese Patent Application No. 2001-379255. In the electrophoresis apparatus according to the present invention, the lid of the electrophoresis tank is provided with a magnet, a reed switch is sandwiched between the connection part of the power supply for electrophoresis and the electrophoresis tank, and the reed switch is operated by the magnet of the lid. Without it, it has the safety of not energizing. In addition, the electrophoretic power switch is equipped with a magnet, and the reed switch in the power supply case is operated by magnetic force, so that the circuit part can be completely sealed in the case. A difficult configuration is realized.

However, the electrophoresis apparatus according to the present invention includes a problem that the operation may become unstable due to variations in performance of magnets used everywhere. In addition, since a reed switch for enhancing safety is attached to the electrophoresis tank, liquid may enter the switch part during washing or injection of a buffer solution, leading to failure. For this reason, it is necessary to pay attention to handling, and it cannot be said that it can be used easily. Further, for example, for observation of DNA, a staining solution suspected of carcinogenicity or toxicity such as ethidium bromide may be added to the buffer solution in the electrode tank before electrophoresis. In this case, with the shape of the conventional electrophoresis tank, when the buffer solution is processed as waste liquid after electrophoresis, the liquid shape during disposal is not constant. There was a possibility that the waste liquid was scattered.

Therefore, the present invention is an extremely compact electrophoresis apparatus that takes advantage of the simplicity of the electric circuit disclosed in Japanese Utility Model Publication No. 63-39639, and has an excellent usability that anyone can easily handle, and is less likely to break down. The object is to devise an electrophoresis apparatus with improved safety.
The safety of the apparatus is ensured by a configuration that cannot be energized without a lid, but the lid and the switch provided in the electrophoresis tank are not interlocked as in the electrophoresis apparatus of Japanese Patent Application No. 2001-379255. And a switch provided in the power supply for electrophoresis. This makes it possible to eliminate switches that are sensitive to liquids from the electrophoresis tank, and dramatically reduce the possibility of failure. Moreover, since it becomes easy to wash | clean an electrophoresis tank, a usability | use_condition improves. In particular, when the buffer solution in the electrode tank is treated as a waste liquid after electrophoresis, it is possible to greatly avoid situations where the waste liquid adheres to the outside of the electrophoresis tank or the waste liquid scatters around.

The present invention is safer, as well as can be realized an efficient electrophoresis experiment, the experimenter, intends smooth line supply of custom-made specific paraphernalia and supplies that need.

The electrophoresis apparatus and the imaging analysis system of the present invention will be described in order.
First, a cross-sectional view of the basic configuration of the electrophoresis apparatus of the present invention is shown in FIG.
The electrophoresis apparatus of the present invention is called a submarine type, and the electrophoresis buffer 2 is injected into the electrophoresis tank 1. The central part of the electrophoresis tank 1 is usually raised in a trapezoidal shape. In addition, by using a small amount of a supporting electrolyte such as a buffer solution used in the electrophoresis tank, it can be diverted to filter paper electrophoresis or cellulose acetate electrophoresis at a place where it rises in a trapezoidal shape. Electrodes 4, 4 ′ are stretched on both bottoms separated by the base 3. The electrodes 4, 4 ' are usually platinum linear members. The two electrodes 4, 4 ' are each connected to a power supply for electrophoresis, one being an anode and the other being a cathode.
An electrophoretic carrier 5 is installed on the platform 3. This electrophoresis carrier 5 is often a gel mainly composed of agarose in a submarine type electrophoresis apparatus. The electrophoresis carrier 5 is provided with a sample installation groove 6 in which the sample 7 is installed. A voltage is applied to the electrophoresis carrier 5 from the electrodes 4 , 4 ′ via the buffer solution 2, and the charged sample 7 is migrated. Each component contained in the sample 7 is separated in the electrophoresis carrier 5 according to a difference in molecular size or a difference in charged state.

In addition, since a high voltage is usually applied in electrophoresis, an electrophoresis tank lid 10 is installed to prevent an electric shock. The electrophoresis tank lid 10 also plays a role of suppressing evaporation of moisture in the buffer solution and preventing contamination such as dust from entering the electrophoresis tank. Further, as shown in FIG. 1A, the partition plates 8a and 8a ′ may be installed above the electrodes 4 and 4 ′ as necessary. This is because at least the lower part is opened so as not to impede energization from the electrodes 4 and 4 ', but from the upper part, the electrodes 4 and 4' can be easily touched to cover the electrodes so as not to break down. Is. The partition plates 8 a and 8 a ′ also function to prevent bubbles generated from the electrodes 4 and 4 ′ from flowing into the central portion of the electrophoresis tank 1. The configuration in which the partition plates 8a and 8a ′ are arranged in the direction perpendicular to the bottom surface of the electrophoresis tank has been used for a long time, such as those described in the column of non-patent literature. In order to influence electrophoresis, it is desirable that the partition plates 8a and 8a ′ are pluggable parts that can be attached and detached as necessary. Further, as shown in FIG. 1 (b), the partition plates 8b and 8b ' may be installed inclined from the vertical direction. This is a structure for preventing bubbles generated from the electrodes 4, 4 ′ from adhering to the partition plate and remaining in the electrophoresis buffer 2. If bubbles remain, dissolution in the electrophoresis buffer 2 is promoted, and there is a possibility that electrophoresis will be affected by an increase in concentration in the solution.

Further, as shown in FIG. 1 (c), bottom partition plates 9 and 9 ′ may be installed. The bottom partition plates 9 and 9 ′ are plates that stand upward from the bottom of the buffer solution tank of the electrophoresis tank 1, and are not necessarily vertical, but the upper limit of the bottom partition plates 9 and 9 ′ is the partition plate (It is formed so as to be positioned above the lower limits of 8a and 8a 'in FIG. 1 (a) or 8b and 8b' in FIGS. 1 (b) and 1 (c). Existence of bottom partition plates 9 and 9 ' Therefore, the possibility of touching the electrodes 4 and 4 ′ is further reduced, so that safety is improved.Because the bottom partition plates 9 and 9 ′ also have a function of physically dividing the buffer solution tank, It is also possible to cause discontinuities in the components and concentrations of the buffer solution, and the partition plates 8a, 8a ', 8b, 8b' and the bottom partition plates 9, 9 'need not be separate. Instead, it may be incorporated into the migration tank as a single part 8c having a cross section as shown in FIG.
For example, the partition plates 8a and 8a ′ are provided with guide grooves on the inner surfaces facing each other in the electrophoresis tank, and are installed in the electrophoresis tank so as to be inserted along the guide grooves.
Other examples of the guide groove include those provided with two continuous convex portions in parallel or those provided with a rail-like body.
The inclination of the partition plates 8b and 8b ′ is preferably 3 ° to 30 ° with respect to the side surfaces 1a and 1a ′ of the electrophoresis tank in that bubbles can be efficiently discharged to the outside. In some cases, it may be an inclination.
The partition plates 8a and 8a ′ need only protrude at least from the buffer solution surface and have a gap and a hole penetrating inside the buffer solution, and the shape thereof is not limited thereto.

  In the electrophoresis experiment, different electrochemical reactions occur between the two electrodes, and generally different amounts of bubbles are generated. For example, an electrolysis reaction occurs in which oxygen is generated at the anode and hydrogen twice as much as that is generated at the cathode. Since the reaction is different between the two electrodes, the buffer solution in the vicinity of the two electrodes has different physical properties with the passage of time, affecting the electrophoresis. Even in the basic configuration as shown in FIG. 1, the circulation of the buffer solution occurs mainly due to the difference in the amount of generated bubbles, but the effect is small. Pumps that forcibly circulate the buffer solution for the purpose of making the physical properties of the buffer solution in the vicinity of both poles are also commercially available, but they are more expensive than the electrophoresis tank and require a lot of work to install, making it easy to use Not suitable for purpose. Therefore, as shown in FIG. 2, a method of configuring the inside for the purpose of circulating the buffer solution is considered. The base part of the electrophoresis tank 1 is used as a gel installation member 3a so that it can be attached and detached as necessary. There is a space between the lower surface of the gel installation member 3 a and the electrophoresis tank 1, and the space is filled with the buffer solution 2. In the basic configuration shown in FIG. 1, the buffer solution path between the electrodes exists only in the upper part of the gel. Therefore, the buffer solutions from both electrodes are pushed and exchanged here, but in the configuration shown in FIG. Since the liquid path exists, the buffer solution 2 is easily circulated.

That is, the buffer solution flows from the cathode (electrode with a large amount of bubble generation) 4a to the anode 4b (electrode with a small amount of bubble generation) at the top of the gel, and vice versa at the bottom. Circulate. Since the driving force for circulating the buffer solution is due to the generation of bubbles, the path constituting portion 3b extends from the gel installation member 3a toward the cathode 4a, and further, the bubble guiding portion 3c is formed above the cathode 4a. Yes. Due to the presence of the bubble guiding portion 3c, the bubbles generated from the cathode 4a are guided upward and are efficiently used as a driving force for circulating the buffer solution 2. The narrower the gap between the side surface of the electrophoresis tank 1 and the bubble guiding portion 3c, the more concentrated the flow of bubbles and the greater the driving force. However, since it affects the formation of the electric field, it cannot be made extremely narrow. To the extent. The path | route structure part 3b is for concentrating the driving force by a bubble on the force which draws in a buffer solution from an anode side. In addition, since the gel installation member 3a can be attached in the reverse direction as long as it is detachable, it can cope with the case where the amount of bubbles generated in both electrodes is different. Since the buffer solution bottom path 1A below the gel installation member 3a is energized, if this path is too large, conversely, consumption of the buffer solution is promoted. The diameter of the buffer solution bottom path 1A may be about 3 mm or less in height and about 400 mm ² or less in cross-sectional area, and the width on the cathode 4a side (side on which the buffer solution is sucked by bubbles) may be narrower than the anode 4b side.

The present invention is used in a power supply integrated electrophoresis apparatus for electrophoresis experiments .

The figure which shows one Example of this invention. The figure which shows one Example of this invention.

DESCRIPTION OF SYMBOLS 1 Electrophoresis tank 2 Buffer 3 Base part 3a Gel installation member 3b Path | route component 3c Bubble guide part
4, 4 'electrode 4a electrode (cathode)
4b electrode (anode)
5 Electrophoresis carrier 6 Sample installation groove 7 Sample
8a, 8a 'partition plate
8b, 8b 'partition plate
9, 9 'Bottom divider
10 Electrophoresis tank lid

Claims (2)

A base part (3) raised in a trapezoidal shape is formed in the central part of the electrophoresis tank (1), and electrodes (4) and (4) are formed on both bottoms of the electrophoresis tank (1) divided by the base part (3). 4 '), and the electrophoresis gel (5) is placed on the platform (3), and the electrophoresis is composed of a buffer solution (2) arranged so that at least the electrophoresis gel is immersed. In the migration tank for apparatus, the partition plate (8b) (8b ') is installed by being inserted along the guide groove provided on the opposing inner surface of the migration tank, and installed above the electrodes (4) (4'). The partition plates (8b) and (8b ′) are arranged so as to protrude from the buffer solution (2) and from 3 ° to the side surfaces (1a) and (1a ′) of the electrophoresis tank (1). in 30 °, submarine type electrophoresis apparatus for electrophoresis which is inclined from the bottom of the electrophoresis tank (1) as widens upward .   The upper limit of the bottom partition plates (9) and (9 ') at the position closer to the electrophoresis gel (5) than the inclined partition plates (8b) and (8b') from the bottom of the electrophoresis tank (1) upward. The electrophoresis tank for electrophoretic devices according to claim 1, further comprising a bottom partition plate (9) (9 ') extending to a position above a lower limit of the partition plates (8b) (8b').

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