Electrophoresis device and electrophoresis method

Abstract

The present invention prevents air bubbles generated in a lower second electrode (4b) from entering a first gel flow path (2). The electrophoresis device (1) is provided with a first electrode (4a) as a cathode and a second electrode (4b) as an anode, and is used for vertical electrophoresis in which the first electrode (4a) is arranged on the upper part and the second electrode (4b) is arranged on the lower part, and is provided with: a buffer tank (5) into which a sample is introduced and in which the first electrode (4a) is provided; a first gel flow path (2) connected to the buffer tank (5) and filled with a gel; and a second gel flow path (3) which is connected to the first gel flow path (2), is filled with a gel, and is provided with a first electrode (4a), and in the second gel flow path (3), a second electrode (4b) is provided at a position different from that of the first gel flow path (2).

Description

Technical Field

[0001] The present disclosure relates to an electrophoresis device and an electrophoresis method. Background Art

[0002] A known gel electrophoresis method utilizes the phenomenon that when an electric field is applied to a substance with an electric charge, the substance moves toward an electrode of opposite polarity to analyze biological substances such as nucleic acids and proteins. Typically, electrophoresis gels such as agarose gel and acrylamide gel are used as supports for biological substances. The migration speed in the electrophoresis gel varies depending on the molecular weight of the biological substance, so the biological substance is separated in the form of bands that vary according to the molecular weight. The gel electrophoresis method has a high decomposition ability for the separation of biological substances and is therefore also used to measure the fragment length of DNA to grasp the state of the sample. In addition, by fluorescently labeling DNA and observing its brightness, it is also used for the quantification of DNA of a specific length.

[0003] Widely used electrophoresis methods include disk gel electrophoresis using a glass tube, slab gel electrophoresis using a slab, and submerged electrophoresis using horizontal electrophoresis, all of which are open-type electrophoresis.

[0004] Furthermore, Patent Document 1 discloses a sealed electrophoresis apparatus in which a gel and a buffer are sealed.

[0005] Furthermore, in submerged electrophoresis, the sample is applied to a structure called a well to receive the sample. This causes the sample to diffuse into the well, buffer reservoir, and other areas, resulting in less sharp bands and reduced separation performance. Therefore, to ensure optimal separation performance, vertical electrophoresis is used in sealed electrophoresis devices, rather than horizontal electrophoresis.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: U.S. Patent No. 8,124,029 Summary of the Invention

[0009] Problems to be solved by the invention

[0010] However, in the case of vertical electrophoresis, as Figure 1 As shown, it is known that bubbles 110 generated by electrolysis at the lower electrode 114 b rise, and the bubbles 110 enter the flow path 111 , hindering the conduction of electricity and sometimes causing poor electrophoresis.

[0011] In the case of open electrophoresis, bubbles generated during power application are not a problem. However, in vertical electrophoresis, bubbles 110 generated at the lower electrode 114b can enter the flow path 111, which can be a problem. Since bubbles 110 are generated by electrolysis during electrophoresis, their generation is difficult to avoid.

[0012] Therefore, an object of the present disclosure is to provide an electrophoresis device and an electrophoresis method that can prevent bubbles generated at a lower electrode from entering a flow path.

[0013] Solutions to Problems

[0014] In order to solve the above-mentioned problems, the electrophoresis device disclosed in the present invention comprises a first electrode at the cathode and a second electrode at the anode, and is used for vertical electrophoresis in which the first electrode is arranged at the upper part and the second electrode is arranged at the lower part. The electrophoresis device comprises: a solution tank, into which the sample is introduced and provided with the first electrode; a first gel flow path, which is connected to the solution tank and filled with gel; and a second gel flow path, which is connected to the first gel flow path, filled with gel, and provided with the second electrode. In the second gel flow path, the second electrode is provided at a position different from the axis of the first gel flow path.

[0015] In addition, the electrophoresis device disclosed in the present invention has a first electrode at the cathode and a second electrode at the anode, and is used for vertical electrophoresis in which the first electrode is arranged at the top and the second electrode is arranged at the bottom. The electrophoresis device includes: a solution tank, into which the sample is introduced and provided with a first electrode; a first gel flow path, which is connected to the solution tank and filled with gel; a second gel flow path, which is connected to the first gel flow path, filled with gel, and provided with a second electrode; and a connecting portion, which is connected to the side of the second gel flow path and connects the first gel flow path and the second gel flow path, and the second gel flow path has a protrusion protruding upward than the connecting portion.

[0016] In addition, the electrophoresis method disclosed in the present invention includes: vertically arranging an electrophoresis device in such a manner that the first electrode becomes the upper part and the second electrode becomes the lower part, the electrophoresis device comprising: a solution tank provided with a first electrode; a first gel flow path connected to the solution tank and filled with gel; a second gel flow path connected to the first gel flow path, filled with gel, and provided with a second electrode; and a connecting portion connected to the side of the second gel flow path and connecting the first gel flow path and the second gel flow path, the second gel flow path having a protrusion protruding upward than the connecting portion; introducing a sample into the solution tank; and applying a voltage to the cathode of the first electrode and the anode of the second electrode to cause the sample to be electrophoresed in the gel of the first gel flow path.

[0017] Effects of the Invention

[0018] According to the electrophoresis device and the electrophoresis method of the present disclosure, it is possible to prevent bubbles generated by energization from entering the flow path. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a diagram showing an example of electrophoresis failure caused by air bubbles in a conventional electrophoresis device.

[0020] Figure 2 It is a diagram showing the structure of the electrophoresis device 1 of Example 1.

[0021] Figure 3 It is a diagram showing the structure of the electrophoresis device 1 according to the second embodiment.

[0022] Figure 4 It is a diagram showing the structure of the electrophoresis device 1 of Example 3.

[0023] Figure 5 It is a diagram showing an example of arrangement of electrodes according to an embodiment.

[0024] Figure 6A It is a diagram showing the structure of the electrophoresis device 1 of Example 4.

[0025] Figure 6B It is a diagram showing the structure of another electrophoresis device 1 according to Example 4.

[0026] Figure 7 FIG. 1 is a diagram showing a flow channel device 100 in which a plurality of electrophoresis devices 1 are installed.

[0027] Figure 8 This is a diagram showing the effect of preventing air bubbles from entering the flow channel in the electrophoresis device 1 according to the embodiment. DETAILED DESCRIPTION

[0028] In all figures used to illustrate the present embodiment, parts having the same function are marked with the same reference numerals, and their repeated descriptions may be omitted. In addition, the present invention is not limited to the description of the following embodiment. The present invention is defined by the appended claims, but it is readily understood by those skilled in the art that the specific structure can be modified within the scope of the present invention.

[0029] To facilitate understanding of the invention, the positions, sizes, shapes, and ranges of various structures shown in the drawings and the like may not necessarily represent actual positions, sizes, shapes, and ranges. Therefore, the present invention is not necessarily limited to the positions, sizes, shapes, and ranges disclosed in the drawings and the like.

[0030] In this specification, unless otherwise specified, a constituent element expressed in the singular includes the plural form.

[0031] use Figures 2 to 8 The electrophoresis device 1 according to the embodiment of the present disclosure will be described. Figures 2 to 8 It is a diagram showing the structure of the electrophoresis device 1 according to the embodiment.

[0032] The electrophoresis device 1 includes a first upper electrode 4a and a second lower electrode 4b for applying a voltage. A power supply (not shown) is connected to the first and second electrodes 4a, 4b, enabling voltage to be applied between the pair of electrodes 4a and 4b. Although not shown, a voltage control device for controlling the operation of the power supply may also be connected to the power supply. The electrophoresis device 1 of the embodiment is used in vertical electrophoresis, with the first electrode 4a positioned at the top and the second electrode 4b positioned at the bottom.

[0033] In addition, the following description will be given by taking the case where the sample (target biological substance) is nucleic acid as an example. Alternatively, the target biological substance may be a protein.

[0034] Example 1

[0035] like Figure 2 As shown in (a), the electrophoresis apparatus 1 of Example 1 includes a first gel channel 2, a second gel channel 3 connected to the first gel channel 2, a buffer tank 5 (solution tank) connected to the first gel channel 2, a first electrode 4a provided in the buffer tank 5, and a second electrode 4b provided in the second gel channel 3. A separation medium (gel) for separating a target biological substance is prefabricated (pre-filled) in the first gel channel 2 and the second gel channel 3.

[0036] As gel, for example, known gels such as agarose gel or polyacrylamide gel can be used. Usually, electrophoresis gel is prepared and used before electrophoresis at every turn. However, in the preparation of electrophoresis gel, if it is agarose gel, then it is necessary to dissolve the gel, and if it is a polyacrylamide gel, then it is necessary to adjust the reagents and degas, so it is time-consuming and labor-intensive. Therefore, in the present embodiment, the preparation of electrophoresis gel is not needed, and prefabricated agarose gel or polyacrylamide gel etc. that can be prepared in advance for immediate electrophoresis are used.

[0037] The electrophoresis apparatus 1 includes a buffer tank 5 (solution tank) into which a buffer solution is introduced. This buffer solution is injected into the sample (target biological substance) to be electrophoresed in the gel of the first gel flow channel 2. The first gel flow channel 2 is connected to the buffer tank 5, and the internal space of the buffer tank 5 is spatially continuously connected to the internal space of the first gel flow channel 2. The buffer solution may or may not be pre-prepared.

[0038] The second gel flow path 3 is connected to the first gel flow path 2, and the internal space of the first gel flow path 2 is spatially continuously connected to the internal space of the second gel flow path 3. In addition, the second gel flow path 3 does not have a connection port 3a with the first gel flow path 2 (see Figure 2 Openings other than (b)).

[0039] A first electrode 4a is provided in the buffer tank 5, and a second electrode 4b is provided in the second gel channel 3. The first electrode 4a is a cathode, and the second electrode 4b is an anode.

[0040] The second electrode 4b in the second gel flow path 3 is provided at a position different from the axis 2a of the first gel flow path 2. In other words, the second electrode 4b in the second gel flow path 3 is provided so as not to overlap with the axis 2a of the first gel flow path 2. The axis 2a is a line passing through the center of the first gel flow path 2.

[0041] like Figure 2 As shown in the enlarged view of the second gel flow path 3 in (b), the connection port 3a of the second gel flow path 3 with the first gel flow path 2 is located at a different position within the second gel flow path 3 than vertically above the second electrode 4b. Specifically, the second electrode 4b within the second gel flow path 3 is positioned so as not to overlap with the opening 3b vertically below the connection port 3a. The X and Y directions in the figure represent horizontal directions, and the Z direction represents the vertical direction. In this specification, the X direction is sometimes referred to as the width direction, and the Y direction is sometimes referred to as the depth direction.

[0042] Second electrode 4b in second gel channel 3 is positioned at a different position from axis 2a of first gel channel 2, thereby preventing bubbles generated at second electrode 4b from entering first gel channel 2. This prevents electrophoresis current from being blocked by bubbles generated at second electrode 4b.

[0043] Example 2

[0044] like Figure 3 As shown, the electrophoresis apparatus 1 of Example 2, like the electrophoresis apparatus 1 of Example 1, includes a first gel channel 2, a second gel channel 3 connected to the first gel channel 2, a buffer tank 5 (solution tank) connected to the first gel channel 2, a first electrode 4a provided in the buffer tank 5, and a second electrode 4b provided in the second gel channel 3. The same descriptions as those of Example 1 are omitted as appropriate.

[0045] In Example 2, the first gel flow path 2 is connected to the side surface of the second gel flow path 3. In Example 2, the second electrode 4b in the second gel flow path 3 is also provided at a position different from the axis 2a of the first gel flow path 2. Furthermore, in Example 2, the connection port 3a of the second gel flow path 3 to the first gel flow path 2 is open in the width direction (the X direction in the figure).

[0046] In Example 2, the second electrode 4b in the second gel flow channel 3 is disposed at a position different from the axis 2a of the first gel flow channel 2. This prevents bubbles generated at the second electrode 4b from entering the first gel flow channel 2. This prevents the electrophoresis current from being obstructed by bubbles generated at the second electrode 4b.

[0047] Furthermore, in Example 2, the connection port 3 a opens in the width direction (X direction in the figure), so that compared with Example 1, it is difficult for bubbles to enter the first gel flow path 2 .

[0048] Example 3

[0049] like Figure 4 As shown, the electrophoresis apparatus 1 of Example 3, like the electrophoresis apparatus 1 of Example 1, includes a first gel channel 2, a second gel channel 3 connected to the first gel channel 2, a buffer tank 5 (solution tank) connected to the first gel channel 2, a first electrode 4a provided in the buffer tank 5, and a second electrode 4b provided in the second gel channel 3. The same descriptions as those of Examples 1 and 2 are omitted as appropriate.

[0050] The electrophoresis apparatus 1 of Example 3 includes a connection portion 6 connecting the first gel flow path 2 and the second gel flow path 3. The connection portion 6 is connected to the side surface of the second gel flow path 3 at approximately the center thereof in the vertical direction (the Z direction in the figure). As a result, the second gel flow path 3 has a space (herein referred to as a protrusion 7) that protrudes upward from the connection portion 6 (dashed line in the figure). The second gel flow path 3 of Example 2 has no opening other than the connection port 3a with the connection portion 6.

[0051] The second electrode 4b is provided so that its front end 4c is above the connection portion 6 (dashed line in the figure). Alternatively, the second electrode 4b may be provided so that its front end 4c is below the connection portion 6.

[0052] Since the bubbles generated at the second electrode 4 b are captured by the protrusion 7 , the bubbles generated at the second electrode 4 b do not enter the first gel channel 2 and do not hinder the conduction of electrophoresis.

[0053] Furthermore, since the second electrode 4 b is provided so that the front end 4 c thereof is located above the connection portion 6 , it is possible to prevent bubbles rising from the front end 4 c along the second electrode 4 b from entering the connection portion 6 .

[0054] Other effects are the same as those of Embodiment 1 and Embodiment 2.

[0055] <Direction of Second Electrode 4b>

[0056] Here, the change of the orientation of the second electrode 4b provided in the second gel flow path 3 will be described. The second electrode 4b is not limited to being provided in the lower part of the second gel flow path 3 (see Figure 5 (a)), it can also be set on the upper part of the second gel flow path 3 (refer to Figure 5 (b)) or side (refer to Figure 5 (c)). In addition, Figure 5In the examples (a) to (c) of FIG. 1 , the second electrode 4 b is provided so that the tip 4 c thereof is located closer to the buffer reservoir 5 (upper side) than the connection portion 6 .

[0057] Figure 5 (d) and (e) are side views of the electrophoresis device 1. Figure 5 As shown in (d) and (e) of FIG. 1 , the second electrode 4 b may be provided at a position different from that on the axis 2 a of the first gel flow path 2 in the depth direction (the Y direction in the figure).

[0058] Example 4

[0059] like Figure 6A As shown, the electrophoresis apparatus 1 of Example 4, like the electrophoresis apparatus 1 of Example 1, includes a first gel channel 2, a second gel channel 3 connected to the first gel channel 2, a buffer tank 5 (solution tank) connected to the first gel channel 2, a first electrode 4a provided in the buffer tank 5, and a second electrode 4b provided in the second gel channel 3. The same descriptions as in Examples 1 to 3 are omitted as appropriate.

[0060] like Figure 6A As shown in FIG. 1 ( a ), the electrophoresis apparatus 1 of Example 4 includes a spacer 8 provided between the tip 4 c of the second electrode 4 b and the connection port 3 a of the second gel channel 3 and the first gel channel 2 .

[0061] In addition, if Figure 6A As shown in FIG. 1 ( b ), the electrophoresis apparatus 1 of Example 4 includes a spacer 8 provided between the tip 4 c of the second electrode 4 b and the connection port 3 a of the second gel channel 3 and the connection portion 6 .

[0062] In Example 4, the electrophoresis apparatus 1 includes a spacer 8, which restricts the movement of bubbles generated at the second electrode 4b. This prevents the bubbles from entering the first gel channel 2 and obstructing the electrophoresis current flow. Other effects are the same as those of Example 1.

[0063] In addition, if Figure 6B As in the reference example of FIG, in a structure in which the second electrode 4b in the second gel flow path 3 is arranged to be aligned with the axis 2a of the first gel flow path 2, a partition 8 may be provided between the front end 4c of the second electrode 4b and the connection port 3a. The partition 8 is provided at an angle with respect to the vertical direction so that rising bubbles do not rise directly upward.

[0064] Example 5

[0065] Figure 7 FIG. 1 is a diagram showing a flow channel device 100 in which a plurality of electrophoresis devices 1 are provided. Figure 7As shown, a plurality of electrophoretic devices 1 are provided in the flow path device 100. The electrophoretic device 1 provided in the flow path device 100 may be the electrophoretic device of any one of the above-mentioned embodiments 1 to 4. In embodiment 5, a flow path device 100 provided with the electrophoretic device 1 having the connection portion 6 of embodiment 3 is described. The flow path device 100 includes a power pin 101 connected to the first electrode 4a and a power pin 102 connected to the second electrode 4b. The flow path device 100 includes a power supply for supplying power to the power pins 101 and 102, a power supply control unit for controlling the power supply voltage, and applies a voltage between the first electrode 4a (cathode) and the second electrode 4b (anode).

[0066] like Figure 7 As shown in (a) , a plurality of electrophoresis apparatuses 1 may be arranged such that the connection portion 6 and the first gel channel 2 are provided on the side of the second gel channel 3 in the arrangement direction of the electrophoresis apparatuses 1 (direction A in the figure).

[0067] In addition, if Figure 7 As shown in (b), the plurality of electrophoresis devices 1 may be arranged in such a manner that the connection portion 6 and the first gel flow path 2 are arranged in a direction (direction B in the figure) near the front or deep side relative to the second gel flow path 3. Figure 7 In the example of (b), Figure 7 Compared with (a), the installation space of the connecting portion 6 and the first gel flow path 2 can reduce the size of the flow path device 100.

[0068] <Electrophoresis method>

[0069] Next, an electrophoresis method using the above-mentioned electrophoresis apparatus 1 will be described. Here, an electrophoresis method using the electrophoresis apparatus 1 having the connection portion 6 of Example 3 will be described.

[0070] First, prepare the electrophoresis apparatus 1 described above. For example, as shown in Example 3, the electrophoresis apparatus 1 includes: a solution tank equipped with a first electrode 4a (buffer tank 5); a first gel channel 2 connected to the buffer tank 5 and filled with gel; a second gel channel 3 connected to the first gel channel 2, filled with gel, and equipped with a second electrode 4b; and a connector 6 connected to the side of the second gel channel 3, connecting the first and second gel channels 2 and 3. Gels are pre-formed in the first and second gel channels 2 and 3, as well as the connector 6. Furthermore, the second gel channel 3 has a protrusion 7 that protrudes upward from the connector 6.

[0071] Next, a buffer solution is added to the buffer solution reservoir 5 , and the sample is injected into the buffer solution added to the buffer solution reservoir 5 .

[0072] Finally, a voltage is applied to the cathode of the first electrode 4 a and the anode of the second electrode 4 b to allow the sample to be electrophoresed in the gel of the first gel channel 2 .

[0073] The results of electrophoresis are shown in Figure 8 .like Figure 8 As shown, it was confirmed that the bubbles generated in the second electrode 4 b were captured by the protrusion 7 and did not enter the first gel flow channel 2 .

[0074] Furthermore, the present disclosure is not limited to the aforementioned embodiments and includes various variations. For example, the aforementioned embodiments are described in detail to facilitate understanding of the present disclosure and are not necessarily limited to having all of the described structures. Furthermore, with respect to a portion of the structure of each embodiment, other structures may be added, deleted, or replaced.

[0075] For example, in the above-mentioned embodiment, the electrophoresis apparatus 1 is described in which the gel is pre-formed in the first gel channel 2 and the second gel channel 3 , but the gel may not be pre-formed.

[0076] Furthermore, in the above-described embodiment, the linear electrode 4 b is disposed in the second gel flow channel 3 . However, the shape of the electrode 4 b is not limited to a linear shape, and may be, for example, a T-shaped electrode.

[0077] Explanation of symbols

[0078] 1—electrophoresis device; 2—first gel flow path; 2a—axis; 3—second gel flow path; 3a—connection port; 3b—opening range; 4a—first electrode; 4b—second electrode; 4c—front end; 5—buffer tank; 6—connection portion; 7—protrusion; 8—partition; 100—flow path device; 101, 102—power pins.

Claims

1. An electrophoresis device comprising a first electrode as a cathode and a second electrode as an anode, for vertical electrophoresis in which the first electrode is arranged at an upper portion and the second electrode is arranged at a lower portion. The electrophoresis device is characterized by comprising: a solution tank into which the sample is introduced and provided with the first electrode; a first gel flow path connected to the solution tank and filled with gel; and a second gel flow path connected to the first gel flow path, filled with gel, and provided with the second electrode; The second electrode is provided in the second gel flow path at a position different from that on the axis of the first gel flow path.

2. The electrophoresis device according to claim 1, wherein: The connection port between the first gel flow path and the second gel flow path is provided in the second gel flow path at a position different from that vertically above the second electrode.

3. The electrophoresis device according to claim 1, wherein: The first gel flow path is connected to a side surface of the second gel flow path.

4. The electrophoresis device according to claim 1, wherein: The device further includes a separator provided between the front end of the second electrode and a connection port between the first gel flow path and the second gel flow path.

5. The electrophoresis device according to claim 1, wherein: Gels are prefabricated in the first gel flow path and the second gel flow path.

6. An electrophoresis device comprising a first electrode as a cathode and a second electrode as an anode, for vertical electrophoresis in which the first electrode is arranged at an upper portion and the second electrode is arranged at a lower portion. The electrophoresis device is characterized by comprising: a solution tank into which the sample is introduced and provided with the first electrode; a first gel flow path, connected to the solution tank and filled with gel; a second gel flow path connected to the first gel flow path, filled with gel, and provided with the second electrode; as well as a connecting portion connected to a side surface of the second gel flow path and connecting the first gel flow path and the second gel flow path, The second gel flow path includes a protruding portion that protrudes upward from the connecting portion.

7. The electrophoresis device according to claim 6, characterized in that: The second electrode is provided so that the front end thereof is located above the connecting portion.

8. The electrophoresis device according to claim 6, wherein: The device further includes a separator provided between the tip of the second electrode and a connection port between the connection portion and the second gel flow path.

9. The electrophoresis device according to claim 6, wherein: The second gel flow path has no opening other than the connection port to the connection portion.

10. An electrophoresis method, characterized in that: include: An electrophoresis device is vertically arranged with a first electrode at the top and a second electrode at the bottom, the electrophoresis device comprising: a solution tank provided with the first electrode; a first gel flow path connected to the solution tank and filled with gel; and a second gel flow path connected to the first gel flow path, filled with gel and provided with the second electrode. and a connecting portion connected to a side surface of the second gel flow path and connecting the first gel flow path and the second gel flow path, wherein the second gel flow path has a protrusion protruding upward from the connecting portion; introducing a sample into the solution tank; as well as A voltage is applied to the cathode of the first electrode and the anode of the second electrode to allow the sample to be electrophoresed in the gel of the first gel flow channel.

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