Component for preparing electrophoresis gel and application thereof

Abstract

The present application provides an assembly for the preparation of an electrophoretic gel comprising i) a plurality of hollow tubes (1) open at the top and bottom, and ii) a plurality of bottom caps (2) wherein each bottom cap (2) is configured to seal the lower opening (4) of each hollow tube (1) in a removable form, each hollow tube (1) being made of a transparent and electrically non-conductive plastic thin layer, or a polylactic acid thin layer.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to Chinese patent application number CN 202311237869.3 and filing date September 22, 2023, the entire contents of which are incorporated herein by reference. Field of the Invention

[0003] The present application relates to a component for preparing an electrophoresis gel and a use thereof. Background Art

[0004] Electrophoresis is a commonly used bioanalysis technique that uses the difference in migration rates of nucleic acids or proteins in an electric field to achieve separation, identification and quantification of nucleic acid or protein molecules. Specifically, during electrophoresis, the nucleic acid or protein sample is loaded into the electrophoresis medium, and an electric field is applied to cause the nucleic acid or protein molecules to migrate along the direction of the electric field. Depending on the charge, size, and shape of the nucleic acid or protein molecules, the nucleic acid or protein molecules migrate at different rates to form specific separation bands in the gel or capillary. Due to its convenience, electrophoresis technology is widely used in gene expression research, genetic disease diagnosis, pathogenic microorganism identification, drug screening, etc.

[0005] Gel preparation and gel extraction after electrophoresis are relatively important steps, but there are various inconveniences.

[0006] For example, in the preparation of polyacrylamide (PAGE) gel, it is necessary to assemble glass plates before filling the gel. The glass plates are large in area, and loose assembly will cause gel leakage, affecting the resolution and straightness of the electrophoresis bands. In addition, the glass plates themselves are heavy and easily damaged, and are easily broken or chipped during assembly and cleaning. At the same time, the number of sample wells in the gel is fixed, and the entire piece needs to be electrophoresed together, resulting in great waste and making the cost of gel production high.

[0007] In traditional disc electrophoresis, gel is usually poured into a glass tube, and it is very troublesome to remove the gel after electrophoresis. Usually, a syringe with a long needle is filled with water, and the needle is inserted between the gel and the inner wall of the glass plate. While rotating the glass tube, water is slowly injected to separate the gel strip from the inner wall of the glass tube, and finally the gel strip is blown out with an ear bulb. This is very easy to damage the gel, the efficiency is very low, and it cannot be used on a large scale.

[0008] The biggest problem with agarose gel is that it easily loses water and cracks, has a short shelf life, needs to be made frequently, and is time-consuming and labor-intensive.

[0009] In addition, electrophoresis gels or samples usually contain toxic substances, such as ethidium bromide and polyacrylamide, and gel plates need to be washed intensively afterwards, which is time-consuming and laborious. When cutting and recovering large gels, the strips in different lanes are sometimes close to each other, which can easily cause cross-contamination during cutting. The cut gels are difficult to mark and easy to mix up. Summary of the invention

[0010] In response to the above problems, the inventors of the present application have designed a set of gel-making components, which can i) prepare gels in independent lanes, reduce the possibility of leakage, and avoid cross-contamination of samples; ii) prepare a corresponding amount of gel based on the number of samples, reducing waste; iii) extend the shelf life of the prepared gel; iv) reduce the manufacturing cost and washing burden of the gel-making components; and v) facilitate the recovery of nucleic acid or protein molecules after electrophoresis.

[0011] In a first aspect, the present application provides a component for preparing an electrophoresis gel, which may include i) a plurality of hollow tubes (1) with upper and lower openings, and ii) a plurality of bottom covers (2), wherein each bottom cover (2) may be configured to seal the lower opening (4) of each hollow tube (1) in a detachable manner.

[0012] The size and shape of each hollow tube (1) are completely consistent, and the size and shape of each bottom cover (2) are completely consistent.

[0013] The cross section of the hollow tube (1) may be circular, square, or other suitable shapes. In some embodiments, the cross section of the hollow tube (1) may be circular, and the inner diameter may be about 0.1 cm to 50.0 cm. In some embodiments, the cross section of the hollow tube (1) may be circular, and the inner diameter may be about 0.5 cm to 5.0 cm.

[0014] The hollow tube (1) can be made of a transparent and non-conductive material, such as a transparent and non-conductive plastic. The transparent and non-conductive plastic can be selected from polypropylene (PP), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), polyvinylidene fluoride (PVDF), polyethylene (PE), polycarbonate (PC), polymethyl methacrylate (PMMA), polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), acrylonitrile-styrene copolymer (AS), nylon, polyethylene terephthalate (PET), etc. The transparent and non-conductive material can also be selected from polylactic acid (PLA). The hollow tube (1) can be made of a transparent and non-conductive thin layer material, such as a thin layer material with a thickness of about 0.05mm to 2.00mm, such as a plastic thin layer or a polylactic acid thin layer. In some embodiments, the hollow tube (1) can be composed of a transparent, non-conductive and heat-resistant plastic thin layer or a polylactic acid thin layer. In some embodiments, the hollow tube (1) may be made of polypropylene (PP), polyvinyl chloride (PVC), or polylactic acid (PLA).

[0015] The length of the hollow tube (1) can be 5.0 cm to 100.0 cm, for example, more than 5.0 cm, more than 6.0 cm, more than 7.0 cm, more than 8.0 cm, more than 9.0 cm, more than 10.0 cm, more than 11.0 cm, more than 12.0 cm, more than 13.0 cm, more than 14.0 cm, or more than 15.0 cm, and less than 100.0 cm, less than 90.0 cm, less than 80.0 cm, less than 70.0 cm, less than 60.0 cm, less than 50.0 cm, less than 40.0 cm, less than 30.0 cm, or less than 20.0 cm.

[0016] The hollow tube (1) may include graduations on its surface, for example graduations for marking the height measured from the bottom.

[0017] The shape and size of the bottom cover (2) can be configured to match the shape and size of the lower opening (4) of the hollow tube (1).

[0018] The bottom cover (2) can be made of plastic, silicone rubber, rubber, silicone, polyurethane, metal, wood, glass, ceramic and other materials. In one embodiment, the bottom cover can be made of silicone rubber, rubber, silicone, or plastic. In some embodiments, the bottom cover can be made of silicone rubber.

[0019] In some embodiments, the bottom cover (2) may include a plane portion (5), and a flange portion (6) protruding at a perpendicular angle to the plane portion (5) at the edge of the plane portion (5), wherein the inner diameter of the flange portion (6) matches the outer diameter of the lower opening (4) of the hollow tube (1). The height of the flange portion (6) on the bottom cover (2) may be about 1.0 cm to 3.0 cm. In some embodiments, the height of the flange portion (6) on the bottom cover (2) may be about 1.0 cm. The thickness of the flange portion (6) on the bottom cover (2) may be about 0.1 cm to 0.3 cm. In one embodiment, the bottom cover may be made of silicone rubber, rubber, or silicone.

[0020] In some embodiments, the bottom cover (2) may include a plane portion (7), and an upper protrusion (8) protruding at a perpendicular angle to the plane portion (7) at the center of the plane portion (7), wherein the outer diameter of the upper protrusion (8) matches the inner diameter of the lower opening (4) of the hollow tube (2). The height of the upper protrusion (8) on the bottom cover (2) may be about 1.0 cm to 3.0 cm. In one embodiment, the bottom cover may be made of silicone rubber, rubber, silicone, polyurethane, wood, plastic, ceramic, glass, or metal.

[0021] Each bottom cover (2) may be independent, or may be arranged on a plane in an integrally formed or spliced ​​manner. In one embodiment, a plurality of bottom covers (2) may be made in an integrally formed manner. In one embodiment, a plurality of bottom covers (2) may be arranged on a bottom plate or a base. In one embodiment, a plurality of bottom covers (2) may be arranged on a bottom plate or a base in an integrally formed manner.

[0022] The component for preparing electrophoresis gel of the present application may also include more than one group of top covers (9), each group includes multiple top covers (9), and each top cover (9) can be configured to close the upper opening (3) of each hollow tube (1) in a detachable form.

[0023] In some embodiments, each top cover (9) may include a plane portion (10), and a lower protrusion (11) protruding at a perpendicular angle to the plane portion (10) at the center of the plane portion (10), wherein the outer diameter of the lower protrusion (11) matches the inner diameter of the upper opening (3) of the hollow tube (1). The height of the lower protrusion (11) of each top cover (9) may be about 0.1 cm to 7.0 cm, such as 0.1 cm, 0.5 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm, 3.0 cm, 3.5 cm, 4.0 cm, 4.5 cm, 5.0 cm, 5.5 cm, 6.0 cm, 6.5 cm or 7.0 cm. In one embodiment, the top cover (9) may be made of silicone rubber, rubber, silicone, polyurethane, wood, plastic, ceramic, plastic, or glass.

[0024] In some embodiments, the assembly for preparing electrophoresis gel of the present application may include two sets of top covers (9), wherein the height difference of the lower protrusions (11) of the two sets of top covers (9) is about 0.5 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm, or 3.0 cm. For example, the assembly of the present application may include two sets of top covers (9), and the height of the lower protrusions (11) thereof may be 2.0 cm and 3.0 cm, 2.0 cm and 3.5 cm, 3.0 cm and 4.0 cm, 3.0 cm and 3.5 cm, 4.0 cm and 5.0 cm, 4.0 cm and 5.5 cm, 5.0 cm and 6.0 cm, 5.0 cm and 6.5 cm, etc. In some embodiments, the height difference of the lower protrusions (11) of the two sets of top covers (9) is about 1.0 cm.

[0025] The assembly for preparing electrophoresis gel of the present application may further comprise a bracket for vertically fixing the hollow tube (1). In one embodiment, the bracket is used to vertically fix a plurality of hollow tubes (1) so that the top surfaces or bottom surfaces of the plurality of hollow tubes are at the same horizontal plane.

[0026] In a second aspect, the present application provides a method for preparing an electrophoresis gel using the assembly of the present application, which may include:

[0027] i) taking one or more hollow tubes (1);

[0028] ii) taking a corresponding number of bottom covers (2) to seal the lower openings (4) of the hollow tubes (1) in a detachable manner;

[0029] iii) placing one or more hollow tubes (1) vertically;

[0030] iv) pouring the same amount of gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and then pouring the same amount of distilled water; or

[0031] The same amount of gel solution is poured into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and a corresponding number of top covers (9) are taken to seal the upper opening (3) of each hollow tube (1) in a detachable manner.

[0032] The method may further include, after step iv), v) removing the distilled water or taking out the top cover (9), pouring the same amount of another gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and then pouring the same amount of distilled water; or removing the distilled water or taking out the top cover (9), pouring the same amount of another gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), taking a corresponding number of top covers (9) whose lower protrusions (11) are smaller in height than the lower protrusions (11) of the top cover in step iv), and closing the upper openings (3) of each hollow tube (1) in a detachable manner.

[0033] The present application also protects a hollow tube containing gel and a bottom cover, or containing gel, a bottom cover and a top cover obtained by the method. The hollow tube containing gel can be referred to as a gel tube in the present application.

[0034] In a third aspect, the present application provides an electrophoresis tank for vertical electrophoresis, which comprises i) an upper electrophoresis tank (12), ii) a cathode (13), iii) a lower electrophoresis tank (14), iv) an anode (15), v) a plurality of lane switches (16), and vi) a plurality of connectors (17).

[0035] The upper electrophoresis tank (12) can be a tank body with an open top, located at the top of the electrophoresis tank, and used to accommodate electrophoresis buffer and electrophoresis cathode (13). The lower electrophoresis tank (14) can be a tank body with an open top, located at the bottom of the electrophoresis tank, and used to accommodate electrophoresis buffer and electrophoresis anode (15). The upper electrophoresis tank (12) and the lower electrophoresis tank (14) can be made of non-conductive material, preferably non-conductive transparent material, such as acrylic.

[0036] A plurality of lane switches (16) may be located on the bottom or side of the upper electrophoresis tank (12) and configured to open or close the external channel of the upper electrophoresis tank (12). The plurality of lane switches (16) may control the opening and closing of the external channel of the upper electrophoresis tank (12) independently of each other. In some embodiments, each lane switch (16) may include a valve. Each lane switch (16) may be made of a non-conductive material, such as plastic, glass, ceramic, etc.

[0037] The plurality of connectors (17) can be configured to seal each lane switch (16) and the gel tube. Each connector (17) can be a hollow tube, one end of which is configured to seal with each lane switch (16) and the other end of which is configured to seal with the gel tube. The plurality of connectors (17) can be connected to each lane switch (16) and the gel tube independently of each other. In one embodiment, each connector (17) may be a hollow tube, and includes, from one end to the other end, a lane switch connection hole (22), a first gel tube connection hole (23), a second gel tube connection hole (24), and a third gel tube connection hole (25), wherein the inner diameter of the first gel tube connection hole (23) is smaller than the inner diameter of the second gel tube connection hole (24), the inner diameter of the second gel tube connection hole (24) is smaller than the third gel tube connection hole (25), the inner diameter of the lane switch connection hole (22) is configured to match the outer diameter of each lane switch (16), and the inner diameter of one of the first gel tube connection hole (23), the second gel tube connection hole (24), and the third gel tube connection hole (25) matches the outer diameter of each gel tube. The lane switch connection hole (22), the first gel tube connection hole (23), the second gel tube connection hole (24), and the third gel tube connection hole (25) may have the same outer diameter. In one embodiment, each connector (17) may include a first hollow tube, a second hollow tube, a connecting portion between the first hollow tube and the second hollow tube, and a plurality of hollow tubes with gradually decreasing inner / outer diameters located in the second hollow tube, such as a third hollow tube with an outer diameter matching the inner diameter of the second hollow tube, a fourth hollow tube with an outer diameter matching the inner diameter of the third hollow tube, etc., wherein the first hollow tube is configured to match the shape and size of each lane switch (16), wherein the number of hollow tubes in the second hollow tube can be adjusted to match the shape and size of the gel tube, etc. The plurality of connectors (17) may be made of, for example, rubber or silicone.

[0038] The electrophoresis tank may further include a support plate configured to support and accommodate accessories in the electrophoresis tank. The support plate may include a left side plate (19), a back plate (20), and a right side plate (21), or may only include a left side plate (19) and a right side plate (21). In one embodiment, the support plate may include a guide rail and a guide rail sliding switch on the left side plate (19) and the right side plate (21), or on the left side plate (19), the back plate (20), and the right side plate (21). The support plate may be made of a non-conductive material, such as acrylic.

[0039] The electrophoresis tank may further include a plurality of fixing members (18) configured to vertically fix the gel tube. The plurality of fixing members (18) may fix the gel tube independently of each other. The plurality of gel tube fixing members (18) may be fixed on the left side plate (19) and the right side plate (21) of the electrophoresis tank, or fixed on the back plate (20). The plurality of fixing members (18) may be made of materials such as acrylic and plastic.

[0040] In one embodiment, the upper electrophoresis tank (12) can be configured to slide on the guide rail of the support plate. In one embodiment, the plurality of fixing members (18) can be configured to slide on the guide rail of the support plate.

[0041] In a fourth aspect, the present application provides a method for performing vertical electrophoresis on a sample, comprising:

[0042] i) taking one or more gel tubes obtained according to the second aspect of the present application, and removing the bottom cover (2), or the bottom cover (2) and the top cover (9);

[0043] ii) vertically fixing the one or more gel tubes of step i) in the electrophoresis tank described in the third aspect of the present application;

[0044] iii) adding samples or molecular standards to one or more gel tubes;

[0045] iv) connecting one or more gel tubes to one or more lane switches (16) via one or more connectors (17), and opening the one or more lane switches (16) connected to the gel tubes;

[0046] v) Setting the electrophoresis conditions, turning on the power supply, and starting electrophoresis;

[0047] vi) turning off the power supply, ending the electrophoresis, turning off one or more lane switches (16) connected to the gel tubes, disconnecting one or more gel tubes from one or more connectors (17), and taking out one or more gel tubes;

[0048] vii) optionally, staining and observing, or directly observing, one or more of the gel tubes;

[0049] viii) Optionally, one or more gel tubes are cut to obtain desired gel tube portions.

[0050] Based on the following specific description and examples, other features and advantages of the current disclosure will become clearer, and specific description and examples should not be interpreted as limiting. The contents of all documents, Genbank records, patents and published patent applications cited in this application are expressly included in this article by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] The following detailed description is given by way of example but is not intended to limit the present invention to the specific embodiments described and can be better understood in conjunction with the accompanying drawings.

[0052] Figure 1 A schematic diagram showing the components of the present application for preparing a gel.

[0053] Figure 2A schematic structural diagram of a bottom cover in an assembly according to one embodiment is shown.

[0054] Figure 3 A schematic structural diagram of a bottom cover in an assembly according to another embodiment is shown.

[0055] Figure 4 A schematic diagram showing multiple bottom covers connected together.

[0056] Figure 5 A schematic diagram showing the structure of the top cover in the assembly.

[0057] Figure 6 A front structural schematic diagram of the electrophoresis tank of the present application is shown.

[0058] Figure 7 A schematic diagram of the side structure of the electrophoresis tank of the present application is shown.

[0059] Figure 8 A schematic structural diagram of a connector in an electrophoresis tank according to one embodiment is shown.

[0060] Fig. 9 A schematic diagram showing the connection between the lane switch and the gel tube via a connector.

[0061] The reference numerals in the drawings of the specification are as follows:

[0062] 1-hollow tube, 2-bottom cover, 3-upper opening of hollow tube, 4-lower opening of hollow tube, 5-flat portion of bottom cover, 6-flange portion, 7-flat portion of bottom cover, 8-upper raised portion, 9-top cover, 10-flat portion of top cover, 11-lower raised portion, 12-upper electrophoresis tank, 13-cathode, 14-lower electrophoresis tank, 15-anode, 16-lane switch, 17-connecting piece, 18-fixing piece, 19-left supporting plate, 20-back plate, 21-right supporting plate, 22-lane switch connecting hole, 23-first gel tube connecting hole, 24-second gel tube connecting hole, 25-third gel tube connecting hole. DETAILED DESCRIPTION

[0063] The terms used in this document, unless otherwise specified, have the common meanings in dictionaries, textbooks, technical reference books, or the meanings commonly understood by those skilled in the art. The following descriptions of some terms are only for the purpose of facilitating the understanding of this application, and are not intended to be particularly limited to these terms, unless otherwise specified.

[0064] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0065] "And" herein means "and / or" unless otherwise specified. "Including" and "including" are used interchangeably in this application without limitation.

[0066] The term "hollow tube" refers to a hollow, non-solid tube, including a hollow round tube, square tube or tube of other shapes. "Upper and lower openings" refers to ports that are not closed at both ends of the tube, and the center of the tube is connected to the outside. The "cross section" of a hollow tube refers to the horizontal cross section when the hollow tube is placed vertically. When the cross section of the hollow tube is circular, the "inner diameter" refers to the circular diameter of the inside of the tube along the tube wall, and the "outer diameter" refers to the circular diameter of the outside of the tube along the tube wall. When the cross section of the hollow tube is square, the "inner diameter" refers to the side length of the square along the inside of the tube wall, and the "outer diameter" refers to the side length of the square along the outside of the tube wall. When the cross section of the hollow tube is rectangular, the "inner diameter" refers to the length and width of the rectangle along the inside of the tube wall, and the "outer diameter" refers to the length and width of the rectangle along the outside of the tube wall.

[0067] The “detachable” approach means that the connection between the two objects is not permanent, but can be connected or disconnected as needed.

[0068] "Matching" or "matching" in this application refers to the shape, size, etc. of two objects being in a state that enables the two objects to be tightly combined together in a manner such as engagement, meshing, embedding, etc. For example, the bottom cover "matches" the shape and size of the lower opening of the hollow tube, so that the bottom cover can be tightly combined with the bottom of the hollow tube to achieve the effect of closing the hollow tube. In one embodiment, the inner diameter of the flange portion of the bottom cover "matches" the outer diameter of the lower opening of the hollow tube, so that the flange portion of the bottom cover can tightly wrap the bottom of the hollow tube, thereby achieving the effect of closing the hollow tube. In another embodiment, the outer diameter of the upper protrusion of the bottom cover "matches" the inner diameter of the lower opening of the hollow tube, so that the upper protrusion can be tightly inserted into the hollow tube, thereby achieving the effect of sealing the hollow tube.

[0069] "Sealed" in this application means that two objects are tightly combined so that no gas or liquid can flow from the joint between the two objects. For example, the bottom cover and the hollow tube are tightly combined so that liquid cannot flow out from the joint between the bottom cover and the hollow tube. "Enclosed" means that two objects are relatively tightly combined, but the degree of tightness is weaker than "sealed", and a small amount of gas or liquid can be allowed to flow at the joint between the two objects. For example, the top cover and the hollow tube are relatively tightly combined, and a small amount of liquid is allowed to flow from the joint interface during the combination process.

[0070] In this application, "vertical" refers to a direction perpendicular to the horizontal plane, parallel to the direction of gravity. "Horizontal plane" refers to the plane formed by the surface of water when it is still under the action of gravity, which is perpendicular to the direction of gravity.

[0071] "Vertical electrophoresis" refers to electrophoresis in which the gel is placed vertically and nucleic acid or protein molecules are separated in the vertical direction.

[0072] "Plastic" is a material made by synthesizing or condensing monomer raw materials. It is composed of synthetic resins and additives such as fillers, plasticizers, stabilizers, lubricants, and colorants. Most plastics are light, chemically stable, and will not rust; they have good impact resistance; they have good transparency and wear resistance; they have good insulation and low thermal conductivity; and they generally have good formability and colorability.

[0073] "Polylactic acid", also known as polylactide, is a polyester polymer obtained by polymerization of lactic acid as the main raw material. Polylactic acid is transparent, has good mechanical and physical properties, good thermal stability, and a processing temperature of 170-230°C. Polylactic acid also has good biocompatibility and is biodegradable after use, making it more environmentally friendly than plastics.

[0074] A "thin layer" material, such as a plastic thin layer or a polylactic acid thin layer in the present application, refers to a material with a thickness ranging from about 0.05 mm to 2.00 mm.

[0075] The present application provides a component for preparing electrophoresis gel, which can i) prepare gels of independent swimming lanes to reduce the possibility of leakage; ii) prepare a corresponding amount of gel based on the number of samples to reduce waste; iii) extend the shelf life of the prepared gel; and iv) reduce the manufacturing cost and washing burden of the gel-making component. The gel prepared by the component can be retained in the gel-making tube and used together with the gel-making tube for electrophoresis to avoid cross contamination of samples. In addition, it is also convenient to directly observe the distribution of nucleic acid or protein molecular bands after electrophoresis, and cut and recover specific molecular bands.

[0076] like Figure 1 As shown, the components of the present application may include i) multiple hollow tubes 1 with upper and lower openings, and ii) multiple bottom covers 2, wherein each hollow tube includes an upper opening 3 and a lower opening 4, and each bottom cover may be configured to seal the lower opening of each hollow tube in a detachable form.

[0077] The assembly may include multiple hollow tubes and multiple bottom covers, and a corresponding number of hollow tubes and bottom covers may be used according to the sample to be electrophoresed. In order to maintain the parallelism of the electrophoresis experiment, the size and shape of each hollow tube remain completely consistent, and the size and shape of each bottom cover remain completely consistent.

[0078] The cross section of the hollow tube can be circular, square, or other suitable shapes. That is, the hollow tube can be a round tube, a square tube, or other tubes. Generally speaking, the shape and size of any cross section of the hollow tube are the same, which is convenient for the preparation, use and storage of the hollow tube, and also convenient for the preparation and selection of brackets and fixings for fixing the hollow tube, and connecting parts for connecting the hollow tube.

[0079] The length and thickness of the hollow tube can be determined according to actual conditions. Generally speaking, the length of the hollow tube can be in the range of 5.0 cm to 100.0 cm, particularly in the range of 10.0 cm to 20.0 cm. When the cross section of the hollow tube is circular, its inner diameter can be in the range of about 0.1 cm to 50.0 cm, particularly in the range of about 1.0 cm to 5.0 cm, for example, the inner diameter is 1.5 cm. When the cross section of the hollow tube is square, its inner diameter, i.e. the inner side length, can be in the range of about 0.1 cm to 50.0 cm, particularly in the range of about 1.0 cm to 5.0 cm, for example, 1.0 cm.

[0080] The hollow tube may be made of a transparent and non-conductive material.

[0081] The hollow tube needs to be made of transparent material because it is convenient to observe the amount of gel solution poured in during the gel pouring process and to add samples into the tube. In addition, because the hollow tube containing gel can be directly used for electrophoresis, the selection of transparent material can facilitate the observation of the movement of the sample to be tested in the gel during the electrophoresis process, determine the time to stop the electrophoresis, etc.; and after the electrophoresis is completed, it is convenient to observe the distribution of nucleic acid or protein bands in the instrument, as well as the cutting of specific bands, etc.

[0082] The hollow tube needs to be made of non-conductive material because the hollow tube will be directly added to the electrophoresis tank together with the gel contained therein. The non-conductive material can ensure that the current does not flow through the hollow tube during the electrophoresis process, thereby reducing the heating of the hollow tube and the electrophoresis fluid, as well as the bias of the band migration.

[0083] In some embodiments, the hollow tube needs to be made of heat-resistant material. When the assembly is used to prepare agarose gel, the hollow tube needs to have heat-resistant properties, because the agarose gel needs to be heated at high temperature before pouring and poured into the hollow tube in time, and the hollow tube needs to remain non-shrinking and non-deformed when the high-temperature liquid is poured. When the assembly is used to prepare polyacrylamide gel, the preparation material of the hollow tube is not required to have heat resistance, because polyacrylamide gel can be poured at room temperature.

[0084] In some embodiments, the hollow tube needs to be made of a material that is easy to cut and maintains a certain hardness. The material maintains a certain hardness because the hollow tube needs to maintain a certain shape when preparing the gel and / or using it for electrophoresis, so as not to damage the gel. The material needs to be easy to cut because after electrophoresis, the hollow tube may need to be cut to recover some bands in the gel.

[0085] Thus, based on the above requirements, the hollow tube can be made of transparent, non-conductive, hard, easy to cut, and heat-resistant materials. In some embodiments, the hollow tube can be made of transparent and non-conductive materials. Transparent and non-conductive plastics can be selected from polypropylene (PP), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), polyvinylidene fluoride (PVDF), polyethylene (PE), polycarbonate (PC), polymethyl methacrylate (PMMA), polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), acrylonitrile-styrene copolymer (AS), nylon, polyethylene terephthalate (PET), etc. In some embodiments, the hollow tube can be made of polylactic acid. In particular, the hollow tube can be made of a thin layer of polylactic acid. The thickness of the wall of the hollow tube, in some embodiments, is the thickness of a plastic thin layer or a thin layer of polylactic acid, which can be in the range of about 0.05mm to 2.00mm.

[0086] The surface of the hollow tube may contain scales, such as scales indicating the height from the bottom of the hollow tube. Such scales can help determine the height of the gel liquid poured into the hollow tube, thereby maintaining the consistency of the gel height in multiple hollow tubes. Of course, the consistency of the gel height in multiple hollow tubes can also be maintained by controlling the amount of gel liquid poured into each hollow tube. For example, the same amount of gel liquid can be added to each hollow tube using a measuring cup, a pipette, or a pipette gun. In addition, the presence of the scale also makes it easy to place multiple hollow tubes at the same horizontal height.

[0087] The shape and size of the bottom cover may be configured to match the shape and size of the lower opening of the hollow tube, so that the lower opening of the hollow tube may be sealed after being combined.

[0088] The bottom cover can be made of materials such as plastic, silicone rubber, rubber, silicone, polyurethane, metal, wood, glass, ceramic, etc. In particular, the bottom cover can be made of rubber or silicone.

[0089] In some embodiments, the bottom cover may include a plane portion 5, and a flange portion 6 protruding at a perpendicular angle to the plane portion at the edge of the plane portion, wherein the inner diameter of the flange portion 6 may be configured to match the outer diameter of the lower opening 4 of the hollow tube, so as to seal the lower opening of the hollow tube after the bottom cover is combined with the lower opening of the hollow tube. Figure 2 When the cross section of the hollow tube and the flange are circular, the inner diameter and outer diameter refer to the diameter of the circle; when the cross section of the hollow tube and the flange are square, the inner diameter and outer diameter refer to the side length of the square. The height of the flange may be in the range of about 1.0 cm to 3.0 cm, and the thickness may be in the range of about 0.1 cm to 0.3 cm.

[0090] In some embodiments, the bottom cover may include a plane portion 7, and an upper protrusion 8 protruding from the center of the plane portion at a perpendicular angle to the plane portion, wherein the outer diameter of the upper protrusion 8 matches the inner diameter of the lower opening 4 of the hollow tube, so that the lower opening of the hollow tube is sealed after the bottom cover is combined with the lower opening of the hollow tube. Figure 3 When the cross section of the hollow tube and the upper protrusion are circular, the inner diameter and outer diameter refer to the diameter of the circle; when the cross section of the hollow tube and the upper protrusion are square, the inner diameter and outer diameter refer to the side length of the square. The height of the upper protrusion can be in the range of 1.0 cm to 3.0 cm.

[0091] The multiple bottom covers can exist independently for easy access. Alternatively, the multiple bottom covers can be arranged on a plane in an integrally formed or spliced ​​manner, such as Figure 4 As shown, this allows multiple hollow tubes to be at the same level during gel pouring, making it easier to observe the height of the gel solution.

[0092] The component of the present application may also include more than one group of top covers 9 , each group including a plurality of top covers 9 .

[0093] In order to maintain the parallelism of the electrophoresis experiment, the size and shape of the caps of each group were kept exactly the same.

[0094] One of the functions of the top cover is to seal the upper opening of the hollow tube after the gel is prepared, thereby reducing the evaporation of water in the gel and extending the shelf life of the gel.

[0095] Another function of the top cover is to keep the amount of gel liquid in the multiple hollow tubes consistent. In one embodiment, the top cover may include a flat surface 10 and a lower protrusion 11 protruding from the center of the flat surface at a perpendicular angle to the flat surface, wherein the outer diameter of the lower protrusion 11 is configured to match the inner diameter of the upper opening 3 of the hollow tube, so that the upper opening of the hollow tube can be closed. Figure 5 . Specifically, after pouring an excess (preferably slightly excess) amount of gel liquid into a plurality of hollow tubes, the top cover is slowly put on. During the process of the top cover entering the hollow tube, the excess gel liquid slowly flows out through the gap between the top cover and the hollow tube, thereby maintaining the consistency of the gel liquid in the plurality of hollow tubes. After the gel solidifies, the upper part of the hollow tube is the sample loading area, and the lower part is the gel area. The height of the lower raised portion can be adjusted within a range of about 0.1 cm to 7.0 cm, depending on the length of the hollow tube, etc. The top cover can be made of silicone rubber, rubber, silicone, polyurethane, metal, wood, ceramic, plastic, or glass, etc.

[0096] In the case of preparing agarose gel, the assembly may include only one set of top covers. In the case of preparing polyacrylamide gel, the assembly may include, for example, two sets of top covers, the lower protrusions of the two sets of top covers having different heights, and the height difference depends on the desired thickness of the concentrated gel, for example, about 0.5 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm or 3.0 cm, etc.

[0097] In one embodiment, the hollow tube can be used with only the top cover. That is, after the top cover is put on, the upper opening 3 of the hollow tube is placed vertically with the upper opening 3 facing downward, and gel liquid is added to the lower opening 4 until the liquid level is flush with the lower opening 4, or until a certain scale. In this case, the combination of the top cover and the hollow tube should reach a "sealed" level.

[0098] The assembly may further include a bracket for vertically fixing the hollow tubes. In some embodiments, the bracket may allow the top surfaces or bottom surfaces of the plurality of hollow tubes to be at the same horizontal plane.

[0099] The components of the present application can be used to prepare electrophoresis gel.

[0100] Specifically, the method of preparing an electrophoresis gel using the assembly of the present application may include:

[0101] i) Take one or more hollow tubes as needed;

[0102] ii) taking a corresponding number of bottom covers to seal the lower openings of the hollow tubes in a detachable manner;

[0103] iii) placing one or more hollow tubes vertically;

[0104] iv) pouring the same amount of gel solution into one or more hollow tubes through the upper openings of each hollow tube, and then pouring the same amount of distilled water; or pouring the same amount of gel solution into one or more hollow tubes through the upper openings of each hollow tube, taking a corresponding number of top covers, and closing the upper openings of each hollow tube in a detachable manner.

[0105] The method may further include, after step iv), v) removing the distilled water or taking out the top cover, pouring the same amount of another gel solution into one or more hollow tubes through the upper openings of each hollow tube, and then pouring the same amount of distilled water; or removing the distilled water or taking out the top cover, pouring the same amount of another gel solution into one or more hollow tubes through the upper openings of each hollow tube, taking a corresponding number of top covers whose lower protrusions are smaller in height than the lower protrusions of the top cover in step iv), and closing the upper openings of each hollow tube in a detachable manner.

[0106] Steps i) to iv) of the above method can be used to prepare agarose gel, and steps i) to v) can be used to prepare polyacrylamide gel. When the method does not use a top cover, the amount of gel liquid in multiple hollow tubes can be kept consistent by relying on, for example, scales on the hollow tubes, gel liquid holding tools, etc.

[0107] As mentioned above, the prepared gel can be a polyacrylamide gel or an agarose gel. The concentration of the gel can vary depending on the application.

[0108] Polyacrylamide gel is a gel with a three-dimensional network structure formed by the polymerization of acrylamide and cross-linking agent N,N'-methylenebisacrylamide under the catalysis of riboflavin or ammonium persulfate and tetramethylethylenediamine. Under normal circumstances, the mobility of protein molecules depends on the net charge they carry and the size and shape of the molecules. After the anionic detergent sodium dodecyl sulfate (SDS) is added to the polyacrylamide gel, the mobility of protein molecules mainly depends on the size of the protein, and has nothing to do with the charge and shape. Therefore, non-denaturing polyacrylamide gel electrophoresis can be used to analyze the complex of natural undenatured samples. The protein molecules remain intact during the electrophoresis process and are separated according to charge and molecular weight. In addition, SDS-PAGE can be used to identify the molecular weight and purity of denatured proteins and detect their concentration. Polyacrylamide gel can also be used to separate nucleic acid molecules, such as DNA fragments below 1kb, with a maximum resolution of 1bp. Polyacrylamide gel electrophoresis is currently the most commonly used and highest resolution analysis and identification technology.

[0109] The polyacrylamide gel usually comprises a concentrated gel on the upper layer and a separation gel on the lower layer. The separation gel can be poured into a hollow tube in, for example, step iv) of the above method, and can be a single concentration or a gradient concentration. The concentrated gel can be poured into a hollow tube in, for example, step v).

[0110] The concentration of the stacking gel can be, for example, 4% or 5%. Its main function is to reduce the height difference of the sample when loading, so that all protein molecules can be on the same starting line, reducing problems such as band tailing. The separation gel is a real molecular sieve, which separates proteins of different molecular weights through the three-dimensional pores formed by polyacrylamide. The volume ratio of the separation gel to the stacking gel can be, for example, 5:1, 10:1, or 20:1.

[0111] The concentration of the separation gel should be determined according to the molecular weight of the target protein, and can range from 3% to 30%. When the molecular weight of the protein and nucleic acid is small, the concentration of polyacrylamide can be higher; when the molecular weight of the protein or nucleic acid is large, the concentration of polyacrylamide can be smaller. In some embodiments, the polyacrylamide gel can be a single concentration, such as 8%, 10%, 12% or 16%. The separation gel can also present a concentration gradient distribution from top to bottom, with the concentration close to the concentrated gel being lower, and the concentration gradually increasing as the distance from the concentrated gel increases, so that molecules with little difference in molecular weight can be better separated in increasing concentrations. In some embodiments, the concentration of the polyacrylamide gel can be a gradient change, such as 4-12%, 4-20% or 8-16%. When preparing a gradient polyacrylamide gel, a gradient former can be used in step iv) of the above method. For example, two concentrations of gel solution are configured, mixed and stirred in a gradient mixer, and poured into a hollow tube.

[0112] Agarose is separated from agar and is composed of 1,3-linked pyranose bD-galactose and 1,4-linked 3,6-dehydropyranose aL-galactose, forming a long chain with a relative molecular weight of 104-105. After agarose is heated and dissolved, the molecules are randomly distributed in a coiled shape. When the temperature is lowered, the hydroxyl groups on the sugar molecules between the chains are connected by hydrogen bonds to form a pore structure. The size of the pore size varies with the concentration of agarose. Nucleic acid or protein molecules will encounter resistance when passing through the network structure of agarose gel, and large molecules will encounter greater resistance when surging. DNA is an alkaline substance and carries a negative charge in an alkaline environment such as pH 8. The negative charge on the DNA chain increases with the increase of DNA molecular weight. The charge-to-mass ratio is a constant, so the separation of DNA in electrophoresis is similar to the molecular sieve effect. The size of DNA fragments that can be separated by agarose gel electrophoresis varies depending on the gel concentration. Gels with a gel concentration of 0.5-0.6% can separate DNA fragments of 20bp-50kb. The pore size of agarose gel is too large for protein molecules, and its molecular sieving effect is negligible for most protein molecules. It is now widely used in nucleic acid research.

[0113] Agarose gel usually contains only one layer of separation gel, and the concentration can vary from 0.4% to 4%. In some embodiments, the concentration of agarose gel can be 1%, 1.5%, 2% or 2.5%, etc. In some embodiments, agarose gel contains an upper layer of concentrated gel and a lower layer of separation gel. The concentration of the concentrated gel can be, for example, 0.5%, and the concentration of the separation gel can be, for example, 1%, 1.5%, 2% or 2.5%, etc. The separation gel can be prepared in step iv) of the above method, and the concentrated gel can be prepared in step v). The volume ratio of the separation gel and the concentrated gel can be, for example, 5:1, 10:1, or 20:1.

[0114] Gels, including polyacrylamide gels and agarose gels, can be divided into denaturing gels and non-denaturing gels. For protein molecules, protein molecules remain active in non-denaturing gels, and their mobility is related to molecular weight, isoelectric point and shape. In denaturing gels, denaturants such as SDS are added to denature protein molecules and carry the same density of negative charges, so that mobility is only related to molecular weight. For DNA, denaturation will break the hydrogen bonds between double strands, so that mobility is only related to molecular weight; in non-denaturing gels, nucleic acid molecules can retain secondary structures, and molecular weight is only a distinguishing factor for mobility. For RNA, because RNA in cells generally exists in a single-stranded form, it will form a variety of secondary structures. If it is run directly on the gel without denaturation, the bands that run out will not be able to truly reflect the size of RNA due to the existence of secondary structures. Therefore, denaturation is often required, such as adding denaturants such as urea; non-denaturing gels are the opposite. In order to study various comprehensive conditions such as the secondary structure, isoelectric point, and molecular weight of the molecule, electrophoresis is performed without adding denaturants. In the embodiments of the present application, the gel may be a denatured gel or a non-denatured gel.

[0115] Using the components of the present application, the gel obtained by the method of the present application can be present in a hollow tube. In other words, the method for preparing the gel of the present application can obtain a hollow tube containing a gel and a bottom cover, or containing a gel, a bottom cover and a top cover, which is called a gel tube. The gel tube can be directly assembled into an electrophoresis device to perform electrophoresis of a sample without separating the gel from the hollow tube.

[0116] The present application provides an electrophoresis tank for vertical electrophoresis, which comprises i) an upper electrophoresis tank 12, ii) a cathode 13, iii) a lower electrophoresis tank 14, iv) an anode 15, v) a plurality of lane switches 16, and vi) a plurality of connectors 17. The electrophoresis tank may also comprise a support plate, and / or a plurality of gel tube fixing members 18, such as Figure 6 and Figure 7 shown.

[0117] The upper electrophoresis tank can be a tank body with an opening at the top, located at the top of the electrophoresis tank, for accommodating electrophoresis buffer and electrophoresis cathode. The lower electrophoresis tank can be a tank body with an opening at the top, located at the bottom of the electrophoresis tank, for accommodating electrophoresis buffer and electrophoresis anode. The upper electrophoresis tank and the lower electrophoresis tank can be made of non-conductive materials to ensure that the current is not conducted from the upper electrophoresis tank or the lower electrophoresis tank during the electrophoresis process. The upper electrophoresis tank and the lower electrophoresis tank are preferably composed of non-conductive transparent materials to facilitate observation of the amount of electrophoresis buffer.

[0118] The upper electrophoresis tank and the lower electrophoresis tank may contain electrophoresis buffer. The electrophoresis buffer in the upper electrophoresis tank and the lower electrophoresis tank may contain the same components or different components.

[0119] During electrophoresis, an electrolytic reaction will occur at the anode and the cathode. For example, an oxidation reaction will occur at the anode and a reduction reaction will occur at the cathode. Long-term electrophoresis will make the anode acidic and the cathode alkaline. The electrophoresis buffer can provide a buffering function so that the pH of the cathode and the anode remains substantially unchanged. Another important function of the electrophoresis buffer is to provide conductivity so that nucleic acids or protein molecules can migrate under the action of an electric field. The electrophoresis buffer can also, for example, provide specific ions such as chloride ions to the concentrated gel. The composition of the electrophoresis buffer and its ionic strength will affect the electrophoretic mobility of nucleic acids or protein molecules, and chemical reactions with the sample should be avoided to change the physicochemical properties of the sample or cause it to lose its biological activity. The electrophoresis buffer can contain EDTA, for example, at a concentration of 1-2 mmol / L, which can chelate divalent cations such as Mg ions to prevent the activation of DNA enzymes during electrophoresis, and to prevent Mg ions from forming precipitation with nucleic acids.

[0120] The electrophoresis tank may include a support plate configured to support and accommodate accessories in the electrophoresis tank. In some embodiments, the support plate may include a left side plate 19 and a right side plate 21. In some embodiments, the support plate may include a left side plate 19, a back plate 20, and a right side plate 21. The support plate may include a guide rail and a guide rail sliding switch, and the upper electrophoresis tank and / or the fixing member may be configured to slide on the guide rail, so as to adjust the position of the upper electrophoresis tank and / or the fixing member according to the length of the gel tube, that is, adjust the spacing distance between the upper electrophoresis tank and the lower electrophoresis tank, and / or the position of the fixing member. The guide rail may be arranged on the left side plate and the right side plate, or on the back plate. The support plate also needs to be made of a non-conductive material to ensure that the current does not flow from the support plate during the electrophoresis process.

[0121] A plurality of lane switches may be located on the side of the upper electrophoresis tank, for example, on the surface opposite to the support back plate, and may be configured to open or close the external channel of the upper electrophoresis tank according to the number of gel tubes in the actual electrophoresis. When the lane switch includes a curved portion and has a relatively small inner diameter, it is easy to cause the problem of residual bubbles due to the small space in the internal channel, thus affecting the electrophoresis effect. Therefore, when designing the lane switch, the inner diameter of the lane switch may be appropriately enlarged to avoid the space in the curved portion being too small, such as making a straight up and down pipeline. In some embodiments, a plurality of lane switches may be located on the bottom surface of the upper electrophoresis tank, and may be configured to open or close the external channel of the upper electrophoresis tank according to the number of gel tubes in the actual electrophoresis.

[0122] The control of multiple lane switches is independent, that is, the switch of the upper electrophoresis tank to the external channel can be conveniently opened or closed according to the number of gel tubes actually used and the positions where they are arranged. In some embodiments, the lane switch comprises a valve. Similarly, the lane switch is made of a non-conductive material, such as plastic, glass, ceramic, etc., to ensure that the current is not conducted from the lane switch during the electrophoresis process. The inner diameter of the lane switch roughly matches the inner diameter of the electrophoresis gel. In general, a thicker electrophoresis gel can use a larger lane switch, so that the electrophoresis rate will not be reduced due to the lower current flux at the lane switch. Specifically, the inner diameter value of the electrophoresis gel cannot exceed twice the inner diameter of the lane switch, otherwise there will be a large loss of electrophoresis rate.

[0123] Before electrophoresis begins, the lane switch needs to be connected to the gel tube. Generally, the connection between the two requires the help of a connector, especially when the shapes and sizes of the two are not particularly matched. In addition, given that the size of the gel tube varies greatly in different situations, the connector can be designed to be connected to gel tubes of various sizes. The end of the connector that is connected to the lane switch can be a single size, or it can be changed according to the diameter of the gel tube. For example, in order to speed up the electrophoresis, a thicker electrophoresis gel tube can use a larger lane switch and connector, so that the electrophoresis rate will not be reduced due to the lower current flux at the lane switch or connector.

[0124] Multiple connectors can be used to connect the lane switch to the gel tube independently of each other, and a corresponding number of connectors can be used according to the number of gel tubes actually used. The connector can be a hollow tube, one end of which is configured to be sealed and connected to the lane switch, and the other end of which is configured to be sealed and connected to the gel tube. Depending on the shape and size of the lane switch cross-section, and the shape and size of the gel tube cross-section, the shape and / or size of the two ends of the connector can be the same or different. Depending on the position of the lane switch and the gel tube, the two end faces of the connector may not be in a straight line. In one embodiment, the two end faces of the connector may present a certain angle, such as a 90° angle, such as Figure 7 shown.

[0125] In one embodiment, each connector may be a hollow tube, and includes a lane switch connection hole 22, a first gel tube connection hole 23, a second gel tube connection hole 24, and a third gel tube connection hole 25 in order from one end to the other end, wherein the lane switch connection hole, the first gel tube connection hole, the second gel tube connection hole, and the third gel tube connection hole have the same outer diameter, the inner diameter of the first gel tube connection hole is smaller than the inner diameter of the second gel tube connection hole, and the inner diameter of the second gel tube connection hole is smaller than the third gel tube connection hole, wherein the inner diameter of the lane switch connection hole is configured to match the outer diameter of the lane switch, and the inner diameter of one of the first gel tube connection hole, the second gel tube connection hole, and the third gel tube connection hole matches the outer diameter of each gel tube. Figure 8 . According to the size of the gel tube actually used, select the first gel tube connection hole, the second gel tube connection hole, and the third gel tube connection hole that match or are relatively close thereto. For the convenience of using the electrophoresis tank, the inner diameter of the gel tube connection hole and the outer diameter of the hollow tube of the assembly of the present application are set to match, that is, the connector is tightly connected to the gel tube, preferably a sealed connection. Fig. 9 A schematic diagram showing the lane switch and the gel tube being sealed and connected via a connector.

[0126] In another embodiment, each connector may include a first hollow tube, a second hollow tube, a connecting portion located between the first hollow tube and the second hollow tube, and a plurality of hollow tubes with gradually decreasing diameters located in the second hollow tube, such as a third hollow tube whose outer diameter matches the inner diameter of the second hollow tube, a fourth hollow tube whose outer diameter matches the inner diameter of the third hollow tube, etc., wherein the first hollow tube is configured to match the shape and size of the lane switch, and the number of hollow tubes in the second hollow tube is adjusted to match the size of the gel tube. The second hollow tube, the third hollow tube, or the fourth hollow tube, etc. are selected according to the size of the gel tube actually used. For the convenience of use of the electrophoresis tank, the inner diameters of the second hollow tube, the third hollow tube, the fourth hollow tube, etc. are set to match the outer diameter of the hollow tube of the present application component, so that the connector is tightly connected to the gel tube, preferably a sealed connection.

[0127] The plurality of fixing members 18 can be configured to fix the gel tube vertically and can be configured to independently control the fixation of the gel tube. The plurality of connecting members can be made of non-conductive materials, such as rubber or silicone, and these materials facilitate a tight connection, preferably a sealed connection. The fixing members can also be made elastic or adjustable to facilitate fixing gel tubes of different diameters.

[0128] The present application also provides a method for performing vertical electrophoresis on a sample, comprising:

[0129] i) taking one or more gel tubes obtained by the gel preparation method of the present application, and removing the bottom cover, or the bottom cover and the top cover;

[0130] ii) fixing the one or more gel tubes of step i) in the vertical electrophoresis tank of the present application via, for example, one or more fixing members;

[0131] iii) adding samples or molecular standards to one or more gel tubes;

[0132] iv) connecting one or more gel tubes to one or more lane switches via one or more connectors, and turning on the one or more lane switches connected to the gel tubes;

[0133] v) Setting the electrophoresis conditions, turning on the power supply, and starting electrophoresis;

[0134] vi) turning off the power supply, ending the electrophoresis, turning off one or more lane switches connected to the gel tubes, disconnecting one or more gel tubes from one or more connectors, and taking out one or more gel tubes;

[0135] vii) optionally, staining and observing, or directly observing, one or more of the gel tubes;

[0136] viii) Optionally, one or more gel tubes are cut to obtain desired gel tube portions.

[0137] Before electrophoresis, the bottom cover, or the bottom cover and the top cover of the gel tube need to be removed, one end of the sample chamber facing upwards, fixed with a fixing in the electrophoresis tank, placed vertically, and the gel tube is inserted into the electrophoresis buffer of the lower electrophoresis tank. In the sample chamber of the gel tube, use a micropipette or a pipette and other tools to add the sample or molecular standard to the sample chamber. After that, connect the connector to the lane switch, and then connect the connector to the gel tube. The role of the lane switch, on the one hand, is as described above, to facilitate the assembly of different numbers of gel tubes in the electrophoresis tank, and on the other hand, to facilitate the independent stop of different samples at any time during the electrophoresis process without affecting other lanes, and also to avoid the positive and negative electrodes being directly connected through the electrophoresis buffer and short-circuiting. In the connection between the connector and the gel tube, the appropriate connection hole in the connector can be selected according to the thickness of the gel tube. For example, an electrophoresis gel with a smaller outer diameter can be inserted into the first gel tube connection hole, an electrophoresis gel with a medium outer diameter can be inserted into the second gel tube connection hole of the connector, and a gel tube with a thicker outer diameter can be inserted into the third gel tube connection hole of the connector.

[0138] After that, slowly open the lane switch connected to the gel tube, so that the electrophoresis buffer in the upper tank slowly fills the lane, connect the cathode and anode wires of the electrophoresis tank to the power supply of the electrophoresis instrument, conduct the current to generate an electric field force, and make the biological molecules, such as nucleic acids or proteins, migrate to the anode of the power supply under the action of the electric field force. The sample will enter the gel from the loading chamber, and under the molecular sieve action of the gel, it will show different swimming speeds according to different molecular weights and other factors. Small molecular weights migrate faster, and large molecular weights migrate slower, showing different electrophoretic bands, thereby achieving separation.

[0139] After the electrophoresis is finished, turn off the lane switch and remove the gel tube from the electrophoresis tank. The gel tube containing the sample or molecular standard can be placed directly under the gel imager, or directly stained and placed for observation after staining. Specifically, when observing, the gel tubes are arranged in order and aligned according to the sample information marked on the gel tubes to ensure that the molecular standard can accurately match the position of the sample band.

[0140] When gel cutting and recovery is required, the gel tube can be placed in a UV imager or a blue light instrument for observation, and the position of the band that needs to be cut and recovered can be marked with a marker pen, and then cut one by one with a surgical blade or a special knife, and the cut gel blocks containing the target bands can be placed in the corresponding marked new sample tubes for the next step of purification and recovery.

[0141] After the experiment, the gel tube can be discarded together with the gel inside. Alternatively, the gel can be roughly separated from the gel tube and the empty tube can be discarded. As a disposable item, the burden of recycling and washing the gel tube can be reduced.

[0142] In addition, a sample loading assembly matching the hollow tube can be provided so that the gel tube of the present application can be used for horizontal electrophoresis. The sample loading assembly can have a shape structure similar to the bottom cover containing the flange portion 6, that is, it includes a plane portion and a flange portion protruding at a vertical angle from the edge of the plane portion to the plane portion, and the inner diameter of the flange portion matches the outer diameter of the upper opening of the hollow tube, wherein the flange portion includes an opening near the plane portion, such as an opening of 0.5 cm long and 0.2 cm wide. The sample loading assembly can be configured to "seal" the upper opening of the hollow tube or gel tube in a detachable manner, and allow the electrophoresis buffer to flow into the hollow tube or gel tube from the opening. The "sealing" here means that the sample loading assembly is tightly connected to the hollow tube gel tube except for the opening portion. Before horizontal electrophoresis, a sample can be added to the upper opening of the gel tube, the sample loading assembly is put on, and the gel tube is fixed to the electrophoresis tank in a way that the opening of the sample loading assembly faces upward.

[0143] Although the present application has been described in conjunction with one or more embodiments, it should be understood that the present application is not limited to these embodiments. The description in this application is intended to cover all variant forms and equivalents, all included in the subject matter and scope of the appended claims. All documents cited in this article are fully incorporated herein by reference.

Claims

1. A component for preparing an electrophoresis gel, comprising i) a plurality of hollow tubes (1) with upper and lower openings, and ii) a plurality of bottom covers (2), wherein each bottom cover (2) is configured to seal the lower opening (4) of each hollow tube (1) in a detachable manner, and each hollow tube (1) is made of a transparent and non-conductive plastic thin layer or a polylactic acid thin layer.

2. The assembly of claim 1, wherein the transparent and non-conductive plastic is selected from polypropylene (PP), polyvinyl chloride (PVC), polyvinylidene chloride (PVDC), polyvinylidene fluoride (PVDF), polyethylene (PE), polycarbonate (PC), polymethyl methacrylate (PMMA), polystyrene (PS), acrylonitrile-butadiene-styrene copolymer (ABS), acrylonitrile-styrene copolymer (AS), nylon or polyethylene terephthalate (PET).

3. The assembly of claim 2, wherein the transparent and non-conductive plastic thin layer or the polylactic acid thin layer has a thickness of 0.05 mm to 2.00 mm.

4. The assembly as claimed in claim 1, wherein the hollow tube (1) is a hollow circular tube with an inner diameter of 0.1 cm to 50.0 cm.

5. The assembly as claimed in claim 1, wherein each bottom cover (2) is made of silicone rubber, rubber, silicone, polyurethane, wood, plastic, ceramic, glass, or metal.

6. The assembly as described in claim 1 further comprises more than one set of top covers (9), wherein each set comprises a plurality of top covers (9), and each top cover (9) is configured to close the upper opening (3) of each hollow tube (1) in a detachable manner.

7. A component as described in claim 6, wherein each top cover (9) comprises a planar portion (10) and a lower protrusion (11), wherein the lower protrusion (11) protrudes from the center of the planar portion (10) at a perpendicular angle to the planar portion (10), and the outer diameter of the lower protrusion (11) is the same as the inner diameter of the hollow tube (1).

8. The assembly according to claim 7, wherein the height of the lower protrusion (11) of each top cover (9) is 0.1 cm to 7.0 cm.

9. The component as described in claim 7 comprises two groups of top covers (9), wherein the height of the lower protrusion (11) of one group of top covers (9) differs from the height of the lower protrusion (11) of the other group of top covers (9) by 0.5 cm, 1.0 cm, 1.5 cm, 2.0 cm, 2.5 cm, or 3.0 cm.

10. The assembly according to claim 1, further comprising a bracket configured to vertically fix the hollow tube (1).

11. A method for preparing an electrophoresis gel using the assembly according to any one of claims 1 to 10, comprising: i) taking one or more hollow tubes (1); ii) taking a corresponding number of bottom covers (2) to seal the lower openings (4) of the hollow tubes (1) in a detachable manner; iii) placing one or more hollow tubes (1) vertically; iv) pouring the same amount of gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and then pouring the same amount of distilled water; or The same amount of gel solution is poured into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and a corresponding number of top covers (9) are taken to seal the upper opening (3) of each hollow tube (1) in a detachable manner.

12. The method of claim 11, further comprising after step iv): Remove the distilled water or take out the top cover (9), pour the same amount of another gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), and then pour the same amount of distilled water; or Remove the distilled water or take out the top cover (9), pour the same amount of another gel solution into one or more hollow tubes (1) through the upper opening (3) of each hollow tube (1), take a corresponding number of top covers (9) whose lower protrusions (11) are smaller in height than the lower protrusions (11) of the top cover in step iv), and seal the upper openings (3) of each hollow tube (1) in a detachable manner.

13. A gel tube obtained by the method according to claim 11 or 12, comprising a hollow tube (1) and a gel inside thereof.

14. An electrophoresis tank for vertical electrophoresis, comprising i) an upper electrophoresis tank (12), ii) a cathode (13), iii) a lower electrophoresis tank (14), iv) an anode (15), v) a plurality of lane switches (16), and vi) a plurality of connectors (17), wherein the plurality of lane switches (16) are configured to open or close the external channel of the upper electrophoresis tank independently of each other, and the plurality of connectors (17) are configured to seal and connect the plurality of lane switches (16) to one or more gel tubes according to claim 13 independently of each other.

15. The electrophoresis tank of claim 14, wherein each lane switch (16) comprises a valve.

16. The electrophoresis tank as claimed in claim 14, wherein each connecting member (17) comprises a hollow tube, one end of which is configured to be sealed and connected to each lane switch (16), and the other end of which is configured to be sealed and connected to the gel tube.

17. The electrophoresis tank as claimed in claim 16, wherein each connecting member (17) comprises, in order from one end to the other end, a lane switch connecting hole (22), a first gel tube connecting hole (23), a second gel tube connecting hole (24), and a third gel tube connecting hole (25), wherein the inner diameter of the first gel tube connecting hole (23) is smaller than the inner diameter of the second gel tube connecting hole (24), and the inner diameter of the second gel tube connecting hole (24) is smaller than the inner diameter of the third gel tube connecting hole (25).

18. The electrophoresis tank of claim 14, further comprising a plurality of fixing members (18) configured to vertically fix one or more gel tubes.

19. The electrophoresis tank as described in claim 14 or 18 further comprises a support plate configured to support and accommodate accessories in the electrophoresis tank, the support plate comprising a guide rail and a guide rail sliding switch, wherein the upper electrophoresis tank (12) and / or the fixing member (18) are configured to slide on the guide rail.

20. A method for performing vertical electrophoresis on a sample, comprising: i) taking one or more gel tubes as claimed in claim 13, and removing the bottom cover (2), or the bottom cover (2) and the top cover (9); ii) vertically fixing the one or more gel tubes of step i) in the electrophoresis tank according to any one of claims 14 to 19; iii) adding samples or molecular standards to one or more gel tubes; iv) connecting one or more gel tubes to one or more lane switches (16) via one or more connectors (17), and opening the one or more lane switches (16) connected to the gel tubes; v) Setting the electrophoresis conditions, turning on the power supply, and starting electrophoresis; vi) Turn off the power supply, end the electrophoresis, turn off one or more lane switches (16) connected to the gel tubes, and disconnect one or more gel tubes from one or more connectors (17).

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