Assembly for gel electrophoresis and electrophoresis comb

Abstract

Described herein are assemblies for gel electrophoresis, including a gel-making cassette and an electrophoresis comb, kits, and methods of use thereof. The gel-making cassette includes a fixed plate and a spacer plate coupled to form a cavity therebetween. The electrophoresis comb includes an elongated body having a first end and a second end; a middle portion connected to the elongated body and extending between the first end and the second end; and a plurality of teeth extending from the middle portion between the first end and the second end. The middle portion has a third end and a fourth end. The middle portion is housed in the cavity. The third end and the fourth end engage inner edges of the cavity. The plurality of teeth is spaced from at least one of the third end and the fourth end. Each tooth has a feature that forms a loading well in an electrophoresis gel that can accommodate a larger sample volume.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 63 / 679,541, filed August 5, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention relates to assemblies and electrophoresis combs for gel electrophoresis. Background Technology

[0004] Gel electrophoresis is a common technique for separating biomolecules such as DNA, RNA, peptides, and proteins. In gel electrophoresis, molecules are separated into bands based on the rate at which they migrate through the filtering gel due to an applied electric field.

[0005] The basic units used in this technology include gels encapsulated in glass tubes, or gels sandwiched as plates between glass or plastic sheets. The gels have an open molecular network structure, with defined pores filled with a conductive buffer solution. These pores are large enough to allow migrating macromolecules to pass through the gel.

[0006] The gel is placed in a chamber in contact with a buffer solution, which provides electrical contact between the gel and the cathode or anode of a power source. A sample containing macromolecules and tracer dyes is placed on top of the gel. A potential is applied to the gel, causing the sample macromolecules and tracer dyes to migrate towards the bottom of the gel. Electrophoresis is stopped just before the tracer dyes reach the ends of the gel. The positions of the separated macromolecular bands are then determined. By comparing the distance a particular band has moved to a known size of tracer dye and macromolecule, the sizes of other macromolecules can be determined.

[0007] Polyacrylamide gels are commonly used media for electrophoresis. Other gels suitable for electrophoresis include agarose gels and starch gels. Polyacrylamide gel electrophoresis (PAGE) is widely popular due to its optical transparency, electroneutrality, and adjustable pore size range.

[0008] The preparation methods for PAGE gels are well known in the industry. See Hames and Rickwood, *Protein Gel Electrophoresis* (2nd ed., Oxford University Press, 1990); Andrews, *Electrophoresis* (2nd ed., Oxford University Press, 1986). Typically, a concentrated stock solution containing acrylamide monomer, a cross-linking agent (such as bisacrylamide), a gel buffer (such as Tris-glycine), and a denaturing agent (such as sodium dodecyl sulfate (“SDS”)) is prepared first. These stock solutions can be stored until the gel is needed. During gel preparation, the stock solution is mixed with water in proportion to the desired final concentration of each component. An oxidizing polymerization initiator (such as ammonium persulfate ((NH4)2(SO4)2; APS)) is added to initiate gel polymerization. Polymerization accelerators (e.g., tetramethylethylenediamine ((TEMED))) are often added to promote rapid polymerization.

[0009] In some cases, a "stacking gel" is used. A stacking gel is a gel poured over a separating gel. Compared to a separating gel, a stacking gel typically has a lower pH (6.8, compared to 8.8) and a lower percentage of polyacrylamide (4%, compared to 8–12%). The purpose of a stacking gel is to concentrate and focus the sample, allowing target molecules (such as proteins) to enter the separating gel as tight bands much smaller than the sample volume.

[0010] Traditional electrophoresis gel molds are mostly made of glass. However, glass has disadvantages such as fragility, difficulty in molding, and high cost. Although it is simpler and more economical to make gel molds using plastic materials through processes such as injection molding, using plastic molds to pour electrophoresis gels may lead to a decrease in the separation of macromolecular bands. This decrease in macromolecular band separation may be due to the fact that macromolecules migrate faster at the contact surface between the gel and the mold than inside the gel. The difference in migration rates between the gel surface and the interior may cause macromolecular bands to exhibit tailing.

[0011] Besides the potential decrease in gel resolution, current methods for preparing and running gels (such as SDS-PAGE gels) have another drawback: gel usability is limited by the sample loading volume. This may require the ability to apply larger sample volumes to the gel. This necessitates preparing thicker gels or designing gels where the well volume increases proportionally to the gel thickness. However, this approach results in a proportionally increased current required for a given electric field strength. Increased current at a given electric field strength leads to increased heat generation within the gel, resulting in decreased resolution and performance. Another problem with thicker gels is that protein band transfer efficiency may decrease in downstream applications such as Western blotting.

[0012] One method to increase the sample loading volume is to increase the depth of the loading wells while maintaining a specific width and gel thickness. However, increasing the loading volume may cause the sample liquid layer above the gel to rise, thereby thickening the initiation zone of the protein bands and reducing resolution. If the height of the stacking gel is increased proportionally, the length of the separating gel will be shortened accordingly, which will also lead to a decrease in resolution within the specially designed cartridge size.

[0013] What is currently needed is a new combination device of gel casting box and electrophoresis comb that meets the following technical parameters: while maintaining a specific gel thickness, its sample well volume is at least twice that of the current method, and at the same time, it ensures that the height of the sample liquid layer above the gel is consistent with the standard specification (specific width and thickness) sample well. Utility Model Content

[0014] One embodiment described herein is an assembly for gel electrophoresis, comprising: a gel casting box including a fixing plate and a separator plate connected to form a cavity between the fixing plate and the separator plate; and an electrophoresis comb including an elongated body having a first end and a second end, a middle portion connected to the elongated body and extending between the first end and the second end, the middle portion having a third end and a fourth end, the third end adjacent to the first end of the elongated body, the fourth end adjacent to the second end of the elongated body, the middle portion being received in the cavity; and a plurality of teeth extending from the middle portion and located between the first end and the second end, the plurality of teeth being spaced apart from at least one of the third end and the fourth end. In one aspect, the plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width and being spaced apart from the third end of the middle portion by a first distance, the first distance being greater than the first width. In another aspect, the plurality of teeth includes a second tooth closest to the fourth end of the middle portion, the second tooth having a second width and being spaced apart from the fourth end of the middle portion by a second distance, the second distance being greater than the second width. In another aspect, the first width and the second width are the same. In another aspect, the first distance and the second distance are the same. In another aspect, each of the plurality of teeth has the same width. In another aspect, the plurality of teeth includes a third tooth located between the first and second teeth, the third tooth having a third width greater than the first width and / or the second width. In another aspect, the third width is greater than twice the first width and greater than twice the second width. In another aspect, each of the plurality of teeth includes a first portion and a second portion located on one side of the first portion opposite to the intermediate portion, the second portion forming an angle with respect to the first portion. In another aspect, the second portion forms an angle of 15 to 25 degrees with respect to the first portion. In another aspect, each of the plurality of teeth includes a third portion located on one side of the second portion opposite to the first portion, the first portion defining a first planar surface, the third portion defining a third planar surface, and the first planar surface being parallel to the third planar surface. In another aspect, the volume of at least one of the plurality of teeth is at least 123 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 50 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 70 μL. In another aspect, the volume of each of the plurality of teeth is at least 70 μL. In another aspect, at least one of the plurality of teeth has a volume of at least 46 μL. In another aspect, each of the plurality of teeth has a volume of at least 46 μL. In another aspect, the first tooth has a volume of at least 50 μL, the second tooth has a volume of at least 50 μL, and the third tooth has a volume of at least 123 μL. In another aspect, the plurality of teeth are spaced 1 to 5 mm from the third end and 1 to 5 mm from the fourth end. In another aspect, the plurality of teeth are spaced 6 to 10 mm from the third end and 6 to 10 mm from the fourth end. In another aspect, the plurality of teeth are spaced 11 to 15 mm from the third end and 11 to 15 mm from the fourth end.

[0015] Another embodiment described herein is an electrophoretic comb for gel electrophoresis, comprising: an elongated body including a first end and a second end; a wall extending at least partially around the periphery of the elongated body; a middle portion connecting the wall and extending between the first and second ends of the elongated body, the middle portion having a third end and a fourth end opposite the third end, the third end adjacent to the first end of the elongated body and the fourth end adjacent to the second end of the elongated body; and a plurality of teeth extending from the middle portion and located between the third and fourth ends, the plurality of teeth being spaced apart from at least one of the third and fourth ends. In one aspect, the plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width and being spaced apart from the third end of the middle portion by a first distance, the first distance being greater than the first width. In another aspect, the plurality of teeth includes a second tooth closest to the fourth end of the middle portion, the second tooth having a second width and being spaced apart from the fourth end of the middle portion by a second distance, the second distance being greater than the second width. In another aspect, the first width and the second width are the same. In another aspect, the first distance and the second distance are the same. In another aspect, each of the plurality of teeth has the same width. In another aspect, the plurality of teeth includes a third tooth located between the first tooth and the second tooth, the third tooth having a third width, and the third width being greater than the first width and the second width. In another aspect, the third width is greater than twice the first width, and the third width is greater than twice the second width. In another aspect, each of the plurality of teeth includes a first portion and a second portion located on one side of the first portion opposite to the intermediate portion, and the second portion forms an angle with respect to the first portion. In another aspect, the second portion forms an angle of 15 to 25 degrees with respect to the first portion. In another aspect, each of the plurality of teeth includes a third portion located on one side of the second portion opposite to the first portion, the first portion defining a first planar surface, the third portion defining a third planar surface, and the first planar surface being parallel to the third planar surface. In another aspect, the volume of at least one of the plurality of teeth is at least 123 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 50 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 70 μL. In another aspect, the volume of each of the plurality of teeth is at least 70 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 46 μL. In another aspect, the volume of each of the plurality of teeth is at least 46 μL. In another aspect, the volume of the first tooth is at least 50 μL, the volume of the second tooth is at least 50 μL, and the volume of the third tooth is at least 123 μL. In another aspect, the plurality of teeth are spaced 1 to 5 mm from the third end and 1 to 5 mm from the fourth end. In another aspect, the plurality of teeth are spaced 6 to 10 mm from the third end and 6 to 10 mm from the fourth end. In another aspect, the plurality of teeth are spaced 11 to 15 mm from the third end and 11 to 15 mm from the fourth end.

[0016] Another embodiment described herein is an assembly for gel electrophoresis, comprising: a gel casting cartridge configured to contain an electrophoretic gel casting solution, the cartridge including a fixing plate and a partition plate coupled to form a cavity between the fixing plate and the partition plate, the fixing plate having an upper edge adjacent to the opening of the cavity; and an electrophoresis comb including: an elongated body having a first end, a second end opposite to the first end, and a channel extending between the first end and the second end; a plurality of teeth extending from the elongated body and located between the first end and the second end; and a wall extending at least partially around the outer periphery of the elongated body, a portion of the wall being configured to engage with the upper edge of the fixing plate. In one aspect, the wall extends from a first side of the elongated body, and the channel extends along a second side of the elongated body opposite to the first side. In another aspect, the wall includes a flange extending between the first end and the second end and a lip extending between the first end and the second end, the flange engaging with the upper edge of the fixing plate. In yet another aspect, a groove is formed between the flange and the lip, and the lip improves accessibility to the elongated body to facilitate removal of the electrophoresis comb from the gel casting cartridge. In another aspect, the elongated body has a bevel near its edge within the groove, and the electrophoretic gel filling solution is configured to flow down the bevel and detach from the electrophoretic comb. In another aspect, the bevel forms a 30-degree angle with respect to the elongated body. In another aspect, the bevel is located on the side of the elongated body opposite to the channel. In another aspect, the fixing plate includes: a fixing plate body, an abutment extending along the fixing plate body near an opening, the abutment having an upper edge that is tapered, and a guide rail extending along the fixing plate body, the guide rail being spaced apart from the abutment and having a tapered edge. In another aspect, the electrophoretic comb includes an intermediate portion located between the elongated body and a plurality of teeth, the intermediate portion extending between a first end and a second end of the elongated body, having a third end and a fourth end opposite to the third end, and the third and fourth ends engaging with the inner edge of the cavity. In another aspect, the channel extends beyond the third and fourth ends. In another aspect, the plurality of teeth are spaced apart from at least one of the third and fourth ends. On the other hand, the assembly also includes a polymerized electrophoretic gel casting solution containing polyacrylamide, one or more buffer solutions, and optional detergents.

[0017] Another embodiment described herein is an electrophoresis comb for gel electrophoresis, comprising: an elongated body including: a first end, a second end opposite to the first end, and a channel extending between the first and second ends; a wall extending at least partially around the periphery of the elongated body, the wall including a flange extending between the first and second ends and a lip extending between the first and second ends; and a plurality of teeth extending from the elongated body between the first and second ends. In one aspect, the wall extends from a first side of the elongated body, and the channel extends along a second side of the elongated body opposite to the first side. In another aspect, a groove is formed between the flange and the lip, wherein the lip improves accessibility to the elongated body. In another aspect, the elongated body has a bevel near the edge within the groove, and the electrophoretic gel pouring solution is configured to flow down the bevel and detach from the electrophoresis comb. In another aspect, the bevel is at a 30-degree angle relative to the elongated body. In another aspect, the bevel is located on the side of the elongated body opposite to the channel. In another aspect, the electrophoretic comb includes an intermediate portion located between an elongated body and a plurality of teeth, the intermediate portion extending between a first end and a second end of the elongated body, and having a third end and a fourth end opposite the third end. In another aspect, a channel extends beyond the third and fourth ends. In another aspect, the plurality of teeth are spaced apart from at least one of the third and fourth ends.

[0018] Another embodiment described herein is an assembly for gel electrophoresis, comprising: a gel casting box formed by a fixed plate and a separator plate connected together to form a cavity therebetween, the fixed plate having an upper edge adjacent to the opening of the cavity; and an electrophoresis comb comprising: an elongated body having a first end and a second end opposite to the first end; a wall extending at least partially along the outer periphery of the elongated body, the wall including a flange extending between the first and second ends, the flange contacting an upper edge of the fixed plate; and a plurality of teeth extending from the wall between the first and second ends of the elongated body. In one aspect, the elongated body includes a channel extending between the first and second ends. In another aspect, the wall extends from a first side of the elongated body, and the channel extends along a second side of the elongated body opposite to the first side. In another aspect, each of the plurality of teeth includes a first portion and a second portion located on a side opposite to the first portion of the wall, the second portion forming an angle with respect to the first portion. In another aspect, the second portion forms an angle of 15 to 25 degrees with respect to the first portion. In another aspect, each of the plurality of teeth includes a third portion located on one side of a second portion opposite to the first portion, the first portion defining a first planar surface, the third portion defining a third planar surface, and the first planar surface being parallel to the third planar surface. In another aspect, the volume of at least one of the plurality of teeth is at least 123 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 50 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 70 μL. In another aspect, the volume of each of the plurality of teeth is at least 70 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 46 μL. In another aspect, the volume of each of the plurality of teeth is at least 46 μL.

[0019] Another embodiment described herein is an electrophoresis comb for gel electrophoresis, configured to be mounted in a gel casting cartridge including an upper edge. The electrophoresis comb includes: an elongated body including a first end and a second end; a wall extending at least partially along a peripheral portion of the elongated body, the wall including a flange extending between the first and second ends for engaging the upper edge of the gel casting cartridge; a middle portion connected to the wall and extending between the first and second ends of the elongated body; and a plurality of teeth extending from the middle portion. In another aspect, the elongated body includes a channel extending between the first and second ends. In another aspect, the wall extends from a first side of the elongated body, and the channel extends along a second side of the elongated body opposite the first side. In another aspect, each of the plurality of teeth includes a first portion and a second portion located on one side of the first portion opposite the middle portion, and the second portion is at an angle relative to the first portion. In another aspect, the second portion forms an angle of 15 to 25 degrees relative to the first portion. In another aspect, each of the plurality of teeth includes a third portion located on one side of a second portion opposite to the first portion, the first portion defining a first planar surface, the third portion defining a third planar surface, and the first planar surface being parallel to the third planar surface. In another aspect, the volume of at least one of the plurality of teeth is at least 123 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 50 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 70 μL. In another aspect, the volume of each of the plurality of teeth is at least 70 μL. In another aspect, the volume of at least one of the plurality of teeth is at least 46 μL. In another aspect, the volume of each of the plurality of teeth is at least 46 μL.

[0020] Another embodiment described herein is a kit comprising: the assembly described herein, a polymeric electrophoresis gel filling solution containing polyacrylamide, one or more buffer solutions and optional detergents; packaging comprising a bag, pouch, plastic container or box; buffer solution; and optional instructions for use.

[0021] Another embodiment described herein is a method for efficiently loading one or more large-volume samples into an electrophoresis gel, the method comprising: providing the assembly described herein; removing the electrophoresis comb from the assembly; and loading one or more samples into one or more sample wells.

[0022] Another embodiment described herein is a method for running multiple samples on an electrophoresis gel without causing striping of samples near the gel edge, the method comprising: providing the assembly described herein; removing the electrophoresis comb from the assembly; introducing one or more samples into one or more wells; and applying an electric current to induce electrophoresis.

[0023] Another embodiment described herein relates to a method for pouring and loading electrophoretic gels that avoids contamination of at least one well in the polymerized gel within a casting cartridge by excess polymerized gel solution. The method includes: providing the assembly described herein; introducing sufficient electrophoretic gel pouring solution to fill the casting cartridge, wherein the electrophoretic gel pouring solution comprises polyacrylamide, one or more buffer solutions, and optionally a detergent, and wherein polymerization of the electrophoretic gel pouring solution has been initiated by adding a polymerization initiator and optionally a polymerization initiator; inserting the electrophoretic comb described herein into a slot; allowing the electrophoretic gel pouring solution to polymerize for a period of time; removing the electrophoretic comb from the slot, wherein after removal, a minimum amount of excess polymerized gel pouring solution remains in or around one or more wells; and loading one or more samples into one or more wells.

[0024] Another embodiment described herein is a method for detecting low-abundance proteins or proteins in diluted samples by loading large sample volumes: providing the assembly described herein; removing the electrophoresis comb from the assembly; introducing one or more samples into one or more wells, wherein the wells have a volume of 40–100 μL; and applying an electric current to induce electrophoresis.

[0025] Another embodiment described herein is sample electrophoresis using the assembly described herein. In one aspect, the sample is a low-abundance protein or a protein in a diluted sample.

[0026] Another embodiment described herein involves using the electrophoresis comb described herein to prepare the electrophoresis gel, wherein the volume of the sample wells formed by the electrophoresis comb is approximately 40–100 μL. Attached Figure Description

[0027] Figure 1 A perspective view of the electrophoresis comb assembled in the gel casting box is shown.

[0028] Figure 2 Showing Figure 1 Side view of the electrophoresis comb and gel casting box.

[0029] Figure 3 Showing Figure 1 Exploded view of the electrophoresis comb and gel casting box.

[0030] Figure 4 Showing Figure 1 Another exploded view of the electrophoresis comb and gel casting box.

[0031] Figure 5 Showing Figure 1 A three-dimensional view of the electrophoresis comb.

[0032] Figure 6 Showing Figure 1 Another three-dimensional view of the electrophoresis comb.

[0033] Figure 7 Showing Figure 1 Side view of the electrophoresis comb.

[0034] Figure 8 Showing Figure 1 Another side view of the electrophoresis comb.

[0035] Figure 9 Showing Figure 7 Enlarged view of the structure of the electrophoresis comb section.

[0036] Figure 10 Showing along Figure 2 A cross-sectional view of the electrophoresis comb and gel casting box obtained by cutting along line 10-10.

[0037] Figure 11 Showing Figure 10 Enlarged view of the electrophoresis comb and gel casting box structure.

[0038] Figure 12 Displays what can be placed Figure 1 A three-dimensional view of another electrophoresis comb in the gel casting box.

[0039] Figure 13 Showing Figure 12 Another three-dimensional view of the electrophoresis comb.

[0040] Figure 14 Showing Figure 12 Side view of the electrophoresis comb.

[0041] Figure 15 Showing Figure 12 Another side view of the electrophoresis comb.

[0042] Figure 16 Showing what can be assembled with Figure 1 A three-dimensional view of another electrophoresis comb in the gel casting box.

[0043] Figure 17 Showing Figure 16 Another three-dimensional view of the electrophoresis comb.

[0044] Figure 18 Showing Figure 16 Side view of the electrophoresis comb.

[0045] Figure 19 Showing Figure 16 Another side view of the electrophoresis comb. Detailed Implementation

[0046] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For example, any nomenclature and techniques described herein related to electrophoresis, biochemistry, molecular biology, or protein and nucleic acid chemistry are common knowledge in the art. In case of any conflict, this specification (including definitions) shall prevail. Exemplary methods and materials are listed below, but similar or equivalent alternatives may be used when implementing or testing the embodiments and schemes described herein.

[0047] As used herein, terms such as “comprising,” “including,” “containing,” “having,” etc., all mean “comprising.” This disclosure also covers other embodiments, whether explicitly stated or not, that may “comprising,” “consistently comprise,” or “comprise” the embodiments or elements described herein. As used herein, “comprising” is an open-ended term and does not exclude the presence of other elements or method steps not explicitly listed. As used herein, “consistently comprise” limits the scope of the claims to the specified materials or steps, and other content that does not substantially affect the basic and novel characteristics of the claimed utility model. As used herein, “comprises” excludes any elements, steps, or components not expressly recited in the claims.

[0048] As used herein, the terms “a,” “an,” “the,” and similar terms used in the context of this disclosure (especially in the context of the claims) should be interpreted to cover both the singular and the plural, unless otherwise stated herein or the context clearly contradicts this. Furthermore, unless otherwise stated, “a,” “an,” or “the” means “one or more.”

[0049] As used in this article, the term "or" can indicate both a parallel relationship (and) and a selective relationship (or).

[0050] As used in this article, the term "and / or" refers to both a parallel relationship (and) and a selective relationship (or).

[0051] As used in this article, the term “substantially” refers to a large or significant degree, but not entirely.

[0052] As used herein, when the terms “about” or “approximately” are applied to one or more target values, they refer to values ​​similar to the stated reference value, or values ​​within an acceptable error range for a particular value as determined by a person skilled in the art, which will depend in part on how the value is measured or determined, such as limitations of the measurement system. In one aspect, the term “about” refers to all possible values ​​(including integer and decimal parts) of the modified numerical value, with an allowable range of variation up to ±10% of that value. Alternatively, according to convention in the art, “about” may also mean within three or more standard deviations. Or, for example, for biological systems or processes, the term “about” may mean a range of variation within a certain order of magnitude of a numerical value, in other embodiments within a range of five times, and in some embodiments within a range of two times. The symbol “~” used herein signifies “about” or “approximately”.

[0053] All ranges disclosed herein include the endpoint discrete values ​​and all integers and fractions within that range. For example, the range 0.1–2.0 includes 0.1, 0.2, 0.3, 0.4, ..., 2.0. If the endpoint values ​​are modified with “about,” the range is extended to include deviations not exceeding ±10% of any value (including the endpoint values) within that range, or within three or more standard deviations, or as defined above for “about.”

[0054] As used herein, the terms “room temperature,” “RT,” or “ambient temperature” refer to the typical temperature in an indoor laboratory environment. On the one hand, a laboratory environment is thermostatically controlled to maintain a substantially constant temperature or within a specific temperature range. On the other hand, “room temperature” refers to a temperature of approximately 15–30°C, including all integer and endpoint values ​​within the specified range. On the other hand, “room temperature” refers to any of the following temperature ranges under standard atmospheric pressure: approximately 15–30°C; approximately 20–30°C; approximately 22–30°C; approximately 25–30°C; approximately 27–30°C; approximately 15–22°C; approximately 15–25°C; approximately 15–27°C; approximately 20–22°C; approximately 20–25°C; approximately 20–27°C; approximately 22–25°C; approximately 22–27°C; approximately 25–27°C; approximately 15°C ± 10%; approximately 20°C ± 10%; approximately 22°C ± 10%; approximately 25°C ± 10%; approximately 27°C ± 10%; ~20°C, ~22°C, ~25°C, or ~27°C.

[0055] As used herein, the terms “control” or “reference” are used interchangeably. A “reference” or “control” level can be a predetermined value or range used as a baseline or benchmark for evaluating measurement results. “Control” also refers to a controlled experiment.

[0056] As used in this article, "PAGE" refers to polyacrylamide gel electrophoresis. "SDS-PAGE" refers to sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis.

[0057] As used herein, "polyacrylamide" refers to a mixture of acrylamide (CH2=CHC(O)NH2) and bisacrylamide (CH2[NHC(O)CH=CH2]2). The percentage of polyacrylamide in the gel casting solution (typically 5-25%) determines the gel porosity. The molecular weight of the biomolecule determines the gel percentage; the larger the molecular weight, the lower the required gel percentage to achieve optimal separation.

[0058] The "electrophoresis gel preparation solution" mentioned in this article refers to the solution used to prepare electrophoresis gels. This solution contains a specific proportion of polyacrylamide, buffer, and water. For SDS-PAGE, the electrophoresis gel preparation solution typically contains 8–12.5% ​​polyacrylamide, Tris-glycine buffer, and sodium dodecyl sulfate (SDS). The solution is gelled by adding a polymerization initiator (such as the oxidant ammonium persulfate (APS)). Polymerization accelerators (e.g., tetramethylethylenediamine (TEMED)) are usually added to promote rapid polymerization. After the addition of the polymerization initiator, the gel polymerizes within 10–30 minutes.

[0059] As used herein, a "stacking gel" refers to an electrophoresis gel consisting of at least two layers: an upper gel with a lower pH and a lower percentage of polyacrylamide, and a lower gel (or "separating gel") with a higher pH and a higher percentage of polyacrylamide. The stacking gel concentrates the sample volume in the wells, allowing the entire sample to enter the separating gel as a focused, narrow band.

[0060] As used herein, “electrophoresis buffer” refers to the buffer solution present at the positive and negative electrodes of the gel. A typical SDS-PAGE electrophoresis buffer (1x concentration) contains 25 mM tris(hydroxymethyl)aminomethane (Tris), 192 mM glycine, pH 8.3, and 0.1% sodium dodecyl sulfate.

[0061] The “electrophoresis gel running apparatus” described herein refers to an assembly comprising: at least one electrophoresis gel assembly, a negative electrode, a positive electrode, a buffer tank with positive and negative electrodes, and a power source for supplying current. In typical electrophoresis gel operation, the negative electrode is located near the sample well (top), and the positive electrode is located near the opposite side (bottom). The electrophoresis gel assembly comprises the gel casting box and polyacrylamide gel described herein.

[0062] This document describes a gel electrophoresis apparatus comprising a gel casting box and an electrophoresis comb with at least one tooth. The gel casting box may include a fixing plate and a separator plate. These plates can be manufactured by processes such as injection molding. Suitable plastic materials for making the plates include, but are not limited to: polyethylene terephthalate, polyvinyl chloride, polymethyl methacrylate, polystyrene, polyethylene, polymethyl polypropylene, and cellulose acetate and copolymers thereof. In some embodiments, a transparent plastic material may be used to allow observation of the gel state within the assembled gel casting box. In some embodiments, the gel casting box may be constructed from a glass plate, or from a glass plate and a plastic plate. In some embodiments, the apparatus may also be configured to prepare pre-formed gels that can be laid flat on various surfaces. This ability to prepare pre-formed gels that can be laid flat is beneficial for product transportation, storage, and daily user operations. Furthermore, during gel production, the gel casting box can be placed flat in an automated loading device.

[0063] In some embodiments, the separator plate has a series of feature structures, protrusions, or pillars along its lateral length, bottom length, or both. These pillars act as energy conductors during assembly, ensuring a uniform seal around the plate. In some embodiments, a slot may be formed at the bottom of the separator plate, and in some embodiments, this slot may optionally penetrate the thickness of the plate. The slot allows the gel inside the gel casting cartridge to contact the buffer solution (such as anolyte buffer) during electrophoresis, thereby forming or assisting in the formation of a circuit pathway. During gel casting, the slot can be temporarily sealed with a sealing material (such as tape, epoxy resin, polymer, conductive polymer, or other suitable sealant) to prevent the gel material from overflowing from the assembled gel casting cartridge before curing. In some embodiments, the separator plate may have flanges that cooperate with spacers on a fixing plate to form gel chambers. In some embodiments, the sealing material may be pre-placed on the separator plate before assembly, or it may be manually applied by the user to the slots of the assembled gel casting cartridge before gel infusion. If a non-conductive sealing material is used, it can be removed after the gel has cured to allow the gel to contact the buffer solution to complete or assist in the completion of the circuit connection. The top surface of the partition plate may be provided with a gripping portion or a curved surface for handling the assembled gel box. In some embodiments, at least one sheet material (e.g., a fixing plate) may have a spacer strip on its inner surface. This spacer strip may take any suitable shape (e.g., typically a U-shaped raised spacer strip on the inner surface). In some embodiments, the spacer strip may be located on the inner surface of one sheet and contact the inner surface of another sheet. In some embodiments, spacer strips may be provided on the inner surfaces of both sheets, and these two spacer strips may contact each other when the box is assembled. An additional function of the spacer strips (single or multiple) is to maintain the spacing between the partition plate and the fixing plate when the sheets are joined to form the box. When the fixing plate and the partition plate are joined, their inner surfaces form a plate-like cavity that can be sealed by the raised spacer strips. In some embodiments, flanges on the partition plate form a seal between the partition plate and the fixing plate. In some embodiments, the spacer strips and flanges together seal the gel chamber. In some embodiments, the fixing plate and the partition plate may be manufactured as a single plastic part. In some embodiments, the fixing plate and the partition plate may be two separate components combined by a connection. These two separate components can be joined by any suitable connection mechanism, including but not limited to, adhesive, pressure fitting, welding, bonding agents, clamps, snap-fits, or any other suitable mechanism (for example only). The thickness of the cavity can be determined by the height of the spacer. In some embodiments, the cavity formed by the spacer and the fixing plate can accommodate a gel material capable of forming a gel of about 1 mm thickness.

[0064] In some embodiments, the abutment extends at an angle from the outer surface of one of the plates to form an opening. Gel material can enter the housing through this opening. Furthermore, structures for forming sample wells in the gel can be introduced into the gel material through this opening. In some embodiments, the abutment may extend at an angle from the outer surface of one of the plates, for example, at an angle of about 70° to a plane perpendicular to the outer surface of the plate, or at an angle of about 20° to the plate surface. In some embodiments, the angle may be between about 15° and about 25°, between about 10° and 30°, or between about 5° and 35°, including all integer angles within the specified range, or other suitable angles. In some embodiments, the angle at which the abutment extends from the outer surface of the plate may be less than about 45°. In some embodiments, the protrusion facilitates the stacking of the welded housings in a storage cassette, which aids in the manufacturing process. In some embodiments, the assembled gel casting cartridge may have at least one abutment on at least one side to form an opening for receiving an electrophoresis comb for forming sample wells in the gel, wherein the electrophoresis comb is wedge-shaped or has a wedge-shaped portion or at least one inclined portion, such as any electrophoresis comb described herein. In some embodiments, abutments may be provided on both sides of the opening to form a V-shaped opening to receive V-shaped teeth on the electrophoresis comb. In some embodiments, the sides of the opening of the cartridge may be flat, but wide enough to receive wedge-shaped or V-shaped teeth on the electrophoresis comb without extending the abutment from one of the substrate surfaces.

[0065] In some embodiments of the device, at least one inner surface of the housing may be coated with a coating. The coating may be an oxygen barrier coating (a coating that forms an oxygen barrier to reduce the diffusion of oxygen from the plastic sheet surface to the gel, thereby preventing significant local variations in the oxygen diffusion rate), for example (by way of example only), including SiO₂. x A coating of SiO or SiO2. In some embodiments, the coating may be substantially transparent. The inner surface may be coated manually or mechanically, and in some embodiments, the coating may be applied before or after the plates are joined together to form a housing when two separate plates are assembled. In some embodiments, the inner surface of the housing may be coated in a vacuum environment by evaporation or vapor deposition, sputtering deposition, chemical vapor deposition, plasma-enhanced chemical vapor deposition, or any combination thereof. In some embodiments, the thickness of the deposited coating may be less than about 5000 Å.

[0066] This paper provides a gel electrophoresis apparatus comprising an electrophoresis comb with a toothed structure configurable to form sample wells, which are configured to hold at least 10% more sample than standard gel sample wells. The increased sample capacity allows more sample to be loaded into the gel without damaging it, while still producing clear, highly separated bands. Furthermore, the larger sample well openings may facilitate sample loading using standard pipette tips (rather than special elongated gel loading tips).

[0067] After the gel material for gel formation is poured into the opening of the assembled gel casting cartridge, an electrophoresis comb can be inserted into the opening and into the gel material. The insertion depth of the electrophoresis comb can be limited by the edge of the comb and the shoulder structure of the cartridge opening. In some embodiments, the insertion depth of the electrophoresis comb can be further limited by the comb abutting against the leading edge of the abutment portion. After the gel is poured, the electrophoresis comb can be removed, leaving a gap in the gel at the position corresponding to each tooth. Each gap forms a sample well with a specific sample volume.

[0068] In some embodiments, the electrophoresis comb may have teeth with beveled surfaces. In some embodiments, the electrophoresis comb may have teeth with wedge-shaped portions. In some embodiments, the electrophoresis comb may have wedge-shaped teeth. Electrophoresis combs with beveled surfaces, wedge-shaped portions, or wedge-shaped teeth can form wedge-shaped sample wells in the gel. In some embodiments, the wedge-shaped teeth have a flat surface at the tooth tip. The flat bottom surface of the teeth forms a sample well with a flat bottom. The flat bottom of the sample well becomes the leading edge of the stacked sample bands, ultimately resolving to straight bands similar to those formed by a standard electrophoresis comb. By inserting an electrophoresis comb with beveled surfaces, wedge-shaped portions, or wedge-shaped teeth into a sample well formed by the gel material, a larger opening is provided at the top of the sample well, making it easier to add samples using standard pipette tips from single-channel or multi-channel pipettes. Furthermore, the sample well allows a larger number of samples to be added to a thinner gel (e.g., a gel about 1 mm thick). In some embodiments, the sample well may be wedge-shaped or contain a wedge-shaped structure. The electrophoretic comb can be used with any of the aforementioned glue-making boxes, or with any other suitable glue-making box.

[0069] The sample wells formed in the gel by teeth on an inclined surface, teeth on a wedge-shaped portion, or wedge-shaped teeth may have a volume at least 12% larger than that formed by a standard electrophoresis comb. In some embodiments, the volume of the sample wells formed by a standard electrophoresis comb is approximately 41 μL. In some embodiments, the sample well volume may be at least 10–200% larger than that formed by a standard electrophoresis comb (inclusive of all integer values ​​within this range); at least 20%, at least 50%, at least 100%, at least 150%, at least 175%, at least 200%, or at least 250% larger. In some embodiments, the volume of the sample wells formed by the electrophoresis comb described herein may be at least 46 μL. In some embodiments, the volume of the sample wells formed by the electrophoresis comb described herein may be approximately 40–200 μL (inclusive of all integer values ​​and sub-ranges within this range). In some embodiments, the volume of the sample wells formed by the electrophoresis comb described herein may be at least approximately 50 μL, at least approximately 70 μL, or at least 123 μL.

[0070] Furthermore, the sample loading wells formed by the electrophoresis comb described herein can increase sample loading while maintaining or improving the sharpness and separation of electrophoretic bands, and reduce edge effects. Sample loading wells formed by inclined surface teeth, wedge-shaped teeth, or wedge-toothed electrophoresis combs allow more sample to be loaded into the wells while keeping the sample away from the sides of the gel. Samples located at the gel edges may not form highly sharp or well-separated bands. Positioning the sample loading wells in such locations reduces the probability of bands forming in the gel exhibiting upward curling ("smile") or downward bending ("frowning") at the ends. Additionally, wedge-shaped or wedge-structured sample loading wells prepared using the apparatus described herein have a larger cross-sectional area at the top than at the bottom. Under constant voltage, current, or power conditions, the increased cross-sectional area results in a lower voltage drop (or electric field strength) at the top of the well compared to the bottom. This lower voltage drop (or electric field strength) in this region results in superior sample stacking, which is better than thinner areas of the well and / or thinner areas of the gel, and also better than gel well designs with uniform thickness throughout the sample loading volume.

[0071] In some embodiments, the medium-sized gel casting cartridge measures approximately 10cm × 10cm. The effective pouring length of the gel loaded within the cartridge is approximately 7cm, and the width is approximately 8cm. The cartridge and the gel poured within it can be of any suitable length. In some embodiments, the width of the cartridge may be designed to reduce edge effects on gels with 10-12 wells. In some embodiments, the assembled cartridge may include a gap extending along the outer periphery of the cartridge. This gap facilitates opening the cartridge to remove the gel. In some embodiments, the cartridge can be opened using a gel scraper or manually.

[0072] In some embodiments, the electrophoresis comb may be a 12-tooth comb, i.e., an electrophoresis comb used to form 12 sample wells in the gel. On one hand, the electrophoresis comb also has an additional smaller tooth at each end (two small teeth in total) for adding control or label samples. On the other hand, the comb teeth are positioned further from the edge, thereby improving the sample filling quality and separation in lanes near the gel edge. In some embodiments, the electrophoresis comb may be a comb with at least one tooth, at least two teeth, at least three teeth, at least four teeth, at least five teeth, at least six teeth, at least seven teeth, at least eight teeth, at least nine teeth, at least ten teeth, at least eleven teeth, at least twelve teeth, at least fourteen teeth, at least fifteen teeth, at least sixteen teeth, at least seventeen teeth, at least eighteen teeth, at least twenty teeth, at least twenty-two teeth, at least twenty-four teeth, at least twenty-six teeth, at least twenty-eight teeth, at least thirty teeth, or any other electrophoresis comb with a suitable number of teeth; in each of the above cases, the electrophoresis comb may also have at least two end teeth smaller than the specified tooth size for loading controls or labeling samples. In some embodiments, the electrophoresis comb may be molded from any suitable plastic, including but not limited to the following polymers: for example, polyethylene terephthalate, polyvinyl chloride, polymethyl methacrylate, polystyrene, polyethylene, polymethyl polypropylene, cellulose acetate, copolymers, polycarbonate, or other suitable materials. In some implementations, the tooth size may remain consistent along the length of the electrophoresis comb or vary along its length. The tooth shape may be the same as or different from the wedge-shaped opening shape of the gel casting box.

[0073] In some embodiments, the edge of the electrophoresis comb opposite the teeth can serve as a handle, facilitating insertion into or removal from the gel casting cartridge. In some embodiments, this edge may be curved. The electrophoresis comb may also include a back and a front. In some embodiments, the front and back are solid structures. In some embodiments, the front of the electrophoresis comb has a recessed space to facilitate the user's gripping of the comb for insertion into the gel casting cartridge and removal from the cartridge after the gel and sample loading wells have formed.

[0074] Glue box

[0075] Figure 1 and Figure 2One embodiment of a gel electrophoresis assembly 100 (also referred to as assembly 100) is shown. The gel electrophoresis assembly 100 includes a gel casting box 104 (also referred to as assembled gel casting box 104) and an electrophoresis comb 108. The gel casting box 104 includes a separator plate 112 and a retaining plate 116 (also referred to as a holding plate 116). The separator plate 112 and the retaining plate 116 can be manufactured by processes such as injection molding. Any suitable plastic can be used to form the separator plate 112 and the retaining plate 116. Suitable plastic materials for forming electrophoretic gel molds include various polymers, such as polyethylene terephthalate, polyvinyl chloride, polymethyl methacrylate, polystyrene, polyethylene, polymethyl polypropylene, cellulose acetate, or various copolymers. In some embodiments, a transparent plastic material may be used to allow observation of the gel state within the assembled gel casting box 104. In some embodiments, the gel casting box 104 may be constructed of a glass plate or a combination of a glass plate and a plastic plate.

[0076] refer to Figure 3 The separator 112 includes an inner surface 120. The separator 112 may have a series of feature structures, protrusions, or pillars 124 along its bottom length and / or side length. The pillars 124 may protrude from the separator 112 and connect to a fixing plate 116. The separator 112 may also have flanges 128 extending along its sides and bottom. These flanges 128 can be used to seal the gel chambers in the assembled gel casting cartridge 104. In some embodiments, the bottom of the separator 112 may have a slot 132. In some embodiments, the slot 132 may optionally extend through the entire thickness of the separator 112. The slot 132 allows the gel inside the gel casting cartridge 104 to contact a buffer solution (such as an anolyte buffer) during electrophoresis, thereby forming or assisting in the formation of circuit pathways. The top surface 136 of the separator 112 may have a gripping portion or a curved surface, for example, to facilitate operation of the assembled gel casting cartridge 104.

[0077] refer to Figure 4 The fixing plate 116 includes an inner surface 140. In some embodiments, the inner surface 140 of the fixing plate 116 may be provided with a generally U-shaped raised spacer 144 that can contact the partition plate 112. For example, the spacer 144 may be accommodated between the post 124 and the flange 128 of the partition plate 112 (e.g., Figure 3 As shown), thereby coupling the fixing plate 116 and the partition plate 112. When the plates are joined to form the glue box 104, the U-shaped spacer 144 can also limit or maintain the spacing between the fixing plate 116 and the partition plate 112. When the partition plate 112 is joined to the fixing plate 116, the inner surfaces 120 and 140 of the two plates 112 and 116 form a plate-shaped cavity 148 (as shown). Figure 11As shown), the cavity is defined by a raised spacer 144. During assembly, it can be accessed via the spacer 144 and the flange 128 on the partition plate 112 (as shown). Figure 3 (As shown) or spacer 144 and flange 128 together seal cavity 148. The thickness of cavity 148 is determined by the height of U-shaped spacer 144. Cavity 148, formed by partition plate 112 and fixing plate 116, can accommodate and confine gel material, which can, for example, form a gel 1 mm thick.

[0078] like Figure 3 As shown, the fixing plate 116 includes an abutment portion 152 extending from the fixing plate 116 at a certain angle, the abutment portion forming an opening 156 leading to the cavity 148 (as shown). Figure 11 (As shown). The abutment portion 152 extends from the outer surface 160 of the retaining plate 116 in a direction away from the partition plate 112 to form an opening 156. The abutment portion 152 includes a first shoulder 164, a second shoulder 168 opposite to the first shoulder 164, and a leading edge 172 located between the two. Both the first shoulder 164 and the second shoulder 168 include a tapered edge 174. The tapered edge 174 of the abutment portion 152 is configured to help limit the abutment portion 152 from damaging the associated storage container. For example, the tapered edge 174 can reduce the risk that the abutment portion 152 may scratch or tear the storage bag or pouch (not shown) of the plastic box 104. The tapered edge 174 may be chamfered, rounded, rounded, or any other suitable shape to prevent tearing, scratching, or other forms of breakage of the storage bag or pouch surrounding the plastic box 104. In some embodiments, the assembled gel casting cartridge 104 may have at least one abutment 152 on at least one side to form an opening 156 for receiving an electrophoresis comb 108 forming a gel loading well. In some embodiments, the separator 112 may also include an abutment 152, thereby making the opening 156 V-shaped.

[0079] Continue to refer to Figure 3 A guide rail 176 may be provided on the outer surface 160 of the fixing plate 116. The guide rail 176 may extend in a direction inclined to the leading edge 172. The guide rail 176 may, for example, facilitate the stacking of multiple glue-making boxes 104 in a storage cassette, thereby assisting in the manufacturing process. Each guide rail 176 includes a tapered guide rail edge 178. The tapered guide rail edge 178 is used to reduce the risk of the guide rail 176 scratching or tearing the storage bag or pouch (not shown) of the glue-making box 104. The tapered guide rail edge 178 may be chamfered, rounded, rounded, or any other suitable shape to prevent tearing, scratching, or other forms of breakage of the storage bag or pouch surrounding the glue-making box 104.

[0080] In some embodiments, the fixing plate 116 and the partition plate 112 may be manufactured as a single plastic part. In some embodiments, the fixing plate 116 and the partition plate 112 may be two separate components joined together. These two separate components may be joined by any suitable connection mechanism, including but not limited to, adhesive, pressure fitting, welding, bonding agents, tape, clamps, snap-fits, or any other suitable mechanism (for example only).

[0081] During gel preparation, the slot 132 on the separator 112 can be sealed. This slot 132 can be sealed with any suitable sealing material (or sealing assembly), including tape, epoxy resin, glue, polymer composites, or other applicable sealants. The sealing material can be conductive or non-conductive. In some embodiments, the sealing material can be a permanent or removable sealant. After the slot 132 is sealed, the gel casting cartridge 104 can be placed vertically, and the gel forming mixture (also known as the electrophoretic gel casting solution) can be poured through the opening 156 into the cavity 148 formed between the retaining plate 116 and the separator 112. After gel casting is complete, a circuit can be formed using a conductive sealing material. Alternatively, the sealant can be removed from the outside of the gel casting cartridge 104, allowing the gel inside the cartridge to contact a buffer solution to complete the circuit path.

[0082] The gel mixture is configured to contact the inner surfaces 120 and 140 of the separator 112 and the fixing plate 116, respectively. In some embodiments, the inner surfaces 120 and 140 of the separator 112 and the fixing plate 116 may be coated. In some embodiments, one or both of the inner surfaces 120 and 140 of the two plates may be coated with an oxygen barrier coating (e.g., SiO2). x A coating (such as SiO or SiO2, or any combination thereof, or other suitable coatings) is applied to form an oxygen barrier, reducing the diffusion of oxygen from the plastic sheet surface to the gel. The oxygen barrier is configured to prevent significant local differences in oxygen diffusion rates. One or more embodiments of the plastic gel mold coating method can be found in U.S. Patent No. 5,685,967, which is incorporated herein by reference in its entirety. In some embodiments, the coating may be substantially transparent. The inner surfaces 120, 140 may be coated manually or mechanically, and in some embodiments, when the gel casting box 104 consists of two separate plates, the coating may be applied before or after the partition plate 112 and the fixing plate 116 are joined to form the gel casting box 104. In some embodiments, the inner surfaces 120, 140 of the gel casting box 104 may be coated under vacuum pressure by evaporation or vapor deposition, sputtering deposition, chemical vapor deposition, plasma-enhanced chemical vapor deposition, or any combination thereof. In some embodiments, the thickness of the deposited coating may be less than about 5000 angstroms.

[0083] In some embodiments, the mini gel casting box 104 is approximately 10cm × 10cm in size. The effective filling length of the gel loaded inside the casting box is approximately 7cm, and the width is approximately 8cm. The casting box 104 and the gel filled inside it can be of any suitable length. In some embodiments, the assembled casting box 104 may include a gap 180 extending along the outer periphery of the casting box 104 (e.g., Figure 1 (As shown). This 180° gap facilitates opening the gel casting box to remove the gel.

[0084] After sample loading onto the gel, the gel casting cartridge 104, containing the sample in the loading well, is placed into the electrophoresis apparatus, which forms a liquid seal between the outer edges of the fixing plate 116 and the separator plate 112. After placement into the electrophoresis apparatus, electrode buffer is added to the anode tank (formed behind the sealed separator plate) and the cathode tank (formed behind the sealed fixing plate). When voltage is applied to the electrophoresis apparatus, anions flow downwards from the cathode tank through the loading well and gel into the anode tank, while cations flow in from the anode tank and through the bottom of the gel. The gel casting cartridge is compatible with any suitable electrophoresis apparatus or system.

[0085] Electrophoretic comb

[0086] After the gel material is poured into the opening 156 of the gel casting cartridge 104, the electrophoresis comb 108 can be inserted into the opening 156. Before inserting the electrophoresis comb 108, a polymerization initiator needs to be added to the gel material to initiate gel polymerization, enabling the gel to be cast or solidified. After the gel solidifies, the electrophoresis comb 108 can be removed, thereby forming multiple pores (or pores or sample loading wells) in the gel corresponding to the teeth of the electrophoresis comb 108. The pores formed by the teeth of the electrophoresis comb 108 are the sample loading wells in the gel. In one aspect, the electrophoresis comb 108 described herein works synergistically with the assembly 100 to prevent excessive polymerized gel from contaminating the sample loading wells and surrounding areas. Furthermore, each electrophoresis comb 108 described herein provides a sample loading well volume larger than that of a standard electrophoresis comb, which allows for the loading of larger volumes of samples for the analysis of low-abundance or diluted protein samples.

[0087] For details, please refer to the following: Figure 5 - Figure 7 The illustration shows an example embodiment of an electrophoresis comb 108. The electrophoresis comb 108 includes an elongated body 184. This elongated body 184 can form a handle so that a user holds the elongated body 184 to insert the electrophoresis comb 108 into the opening 156 of the gel casting cartridge 104 (e.g., ...). Figure 11 (As shown) and / or remove the electrophoresis comb 108 from the gel casting box 104. The elongated body 184 includes a first end 188 and a second end 192 opposite to the first end 188. The elongated body 184 also includes a first side 196 and a second side 200 opposite to the first side 196. The first side 196 and the second side 200 extend between the first end 188 and the second end 192, respectively.

[0088] Continue to refer to Figure 5 - Figure 7 The electrophoresis comb 108 includes a wall 204 extending at least partially along the outer periphery of an elongated body 184. The wall 204 extends from a first side 196 of the elongated body 184 in a direction away from a second side 200. The wall 204 includes a lip 208 (also referred to as a flange 208 or handle 208) and an edge 212. The lip 208 is located on the upper part of the wall 204. In other words, the lip 208 is located on the side of the elongated body 184 facing away from the opening 156 of the gel casting box 104. A user can hold the lip 208 to insert the electrophoresis comb 108 into the opening 156 of the gel casting box 104 and remove the electrophoresis comb 108 from the gel casting box 104. The edge 212 is located on the lower part of the wall 204. In other words, the edge 212 is located on the side of the elongated body 184 adjacent to the opening of the gel casting box 104. Edge 212 engages with the leading edge 172 of the abutment portion 152 on the fixing plate 116 to prevent gel moisture from escaping from the cavity 148. In the illustrated embodiment, wall 204 includes a slot 216. This slot 216 extends through the lip 208. The slot 216 defines a midpoint between the first end 188 and the second end 192. In some embodiments, wall 204 may be without the slot 216, such that wall 204 extends along the entire periphery of the elongated body 184. Wall 204 and first side 196 together define a recess 220. This recess 220 extends between the first end 188 and the second end 192. Recess 220 provides space for a user's fingers to abut against the lip 208 to grip and operate the electrophoresis comb 108.

[0089] like Figure 5 and Figure 11 As best shown, the electrophoresis comb 108 has a bevel 224 within the groove 220. This bevel 224 is located on one side of the groove 220 adjacent to the edge 212. When the electrophoresis comb 108 is inserted into the cavity 148 and contacts the electrophoresis gel casting solution, some solution may overflow the cavity 148 and adhere to the electrophoresis comb 108. The bevel angle of the bevel 224 is sufficient to allow the overflowing electrophoresis gel casting solution to flow down the surface 224 and detach from the electrophoresis comb 108. This angle may be 30 degrees relative to the plane containing the first side 196 of the elongated body 184. In other embodiments, this angle may be approximately 10 degrees, 20 degrees, 40 degrees, 50 degrees, 60 degrees, or any suitable angle (relative to the plane containing the first side 196) to allow the overflowing electrophoresis gel casting solution to flow down the surface 224 and detach from the electrophoresis comb 108.

[0090] Now for reference Figure 6The electrophoresis comb 108 includes a channel 228 (also called a drain channel 228). This channel 228 allows the electrophoresis gel casting solution to flow through, thereby draining from the electrophoresis comb 108 and preventing the electrophoresis gel casting solution from contacting the user. The channel 228 extends into the interior of the second side 200 of the elongated body 184. The channel 228 extends between the first end 188 and the second end 192. The channel 228 is connected to the inclined surface 224 (as shown in the image). Figure 11 (As shown) The slope 224 is designed so that the channel 228 maintains sufficient width to drain the electrophoresis gel casting solution while maintaining proper structural rigidity (or structural integrity) of the electrophoresis comb 108. In the illustrated embodiment, the channel 228 is straight and terminates near the wall 204. In other embodiments, the channel 228 may adopt any suitable shape and location to facilitate the drainage of the electrophoresis gel casting solution from the electrophoresis comb 108.

[0091] refer to Figure 5 - Figure 7 The electrophoresis comb 108 includes a central portion 232. This central portion 232 extends from the wall 204. More specifically, the central portion 232 extends from the edge 212 away from the lip 208. The central portion 232 extends between a first end 188 and a second end 192 of the elongated body 184. The central portion 232 defines a third end 236 and a fourth end 240 opposite to the third end 236. The third end 236 is adjacent to the first end 188 of the elongated body 184, and the fourth end 240 is adjacent to the second end 192 of the elongated body 184. The distance between the third end 236 and the fourth end 240 is less than the distance between the first end 188 and the second end 192. The central portion 232 is configured to be received within a cavity 148 when the electrophoresis comb 108 is inserted into the gel casting cartridge 104. In some embodiments, when the electrophoresis comb 108 is connected to the gel casting box 104, the third end 236 and / or the fourth end 240 may engage with the inner surface of the cavity 148.

[0092] The electrophoresis comb 108 includes a plurality of teeth 248. These teeth 248 extend from the intermediate portion 232. The teeth 248 are configured to form a plurality of voids or pores (or sample loading wells) in the gel of the gel casting cartridge 104. The teeth 248 extend from the intermediate portion 232 away from the elongated body 184. In some embodiments, each tooth 248 may have at least one inclined surface. In some embodiments, each tooth 248 may, for example, be wedge-shaped, include a wedge-shaped portion, or have at least one inclined surface. More specifically, each tooth 248 includes a first portion 252, a second portion 256, and a third portion 260. The first portion 252 is adjacent to the intermediate portion 232. The second portion 256 is located on the side of the first portion 252 away from the intermediate portion 232. The third portion 260 is located on the side of the second portion 256 away from the first portion 252.

[0093] refer to Figure 8 The diagram shows a side view of a single tooth among a plurality of teeth 248 (or a single tooth 248), with a first portion 252 defining a first surface 264 (or a first plane 264), a second portion 256 defining a second surface 268 (or a second plane 268), and a third portion 260 defining a third surface 272 (or a third plane 272). An intermediate portion 232 may further define (or be included in) the first surface 264. The first plane 264 and the third plane 272 are oriented parallel to each other. The second plane 268 is oriented at an angle relative to the first plane 264 and the third plane 272. In other embodiments, the first plane 264 may be oriented at an angle relative to the third plane 272. Furthermore, the second plane 268 may be oriented at an angle 276 relative to the third plane 272. The angle 276 may be 20 degrees. It should be understood that the smaller the angle 276, the longer the tooth 248 can be. In other embodiments, the angle 276 may be any value that allows each tooth 248 to achieve a desired height or length. For example, angle 276 can be 10 degrees, 15 degrees, 25 degrees, 30 degrees, 35 degrees, or greater than 35 degrees. In other embodiments, angle 276 can be any range of values ​​that achieves the desired height or length of each tooth 248. For example, angle 276 can be between 17.5 and 22.5 degrees, between 15 and 25 degrees, between 10 and 30 degrees, etc.

[0094] Continue to refer to Figure 8 The first portion 252 and the intermediate portion 232 together define a first height 278. The value of the first height 278 partially determines the total height 280 of each tooth 248. The larger the total height 280 of each tooth 248, the larger the volume of each tooth 248, the larger the size of the resulting sample dispensing orifice, and the greater the amount of sample that can be dispensed into the corresponding orifice. The first height 278 may be, for example, 1 mm, 2 mm, 5 mm, 10 mm, 15 mm, or any suitable value that achieves the desired volume of the tooth 248. In some embodiments, the first height 278 may be defined solely by the first portion 252.

[0095] refer to Figure 7 The plurality of teeth 248 include: a first tooth 248a adjacent to the third end 236 of the intermediate portion 232, a second tooth 248b adjacent to the fourth end 240 of the intermediate portion 232, and a plurality of third teeth 248c located between the first tooth 248a and the second tooth 248b. The illustrated electrophoresis comb 108 includes 12 third teeth 248c, therefore the electrophoresis comb 108 has a total of 14 teeth 248. In other embodiments, the electrophoresis comb 108 may include any suitable number of third teeth 248c (e.g., 10, 12, 16, etc.). The first tooth 248a and the second tooth 248b have the same structure. The third teeth 248c have a different structure than the first tooth 248a and the second tooth 248b.

[0096] refer to Figure 9 The first tooth 248a has a first width 282a, and the third tooth 248c has a third width 282c. It should be understood that the second tooth 248b has a second width. This second width may be the same as the first width 282a. The third width 282c is greater than the first width 282a and the second width. The third width 282c may, for example, be 2 times, 2.5 times, 3 times, etc., the first width 282a and / or the second width.

[0097] The dimensions of tooth 248 (e.g., first height 278, total height 280, first width 282a, second width, third width 282c, angle 276, etc.) affect the volume of tooth 248. It may be necessary to maximize the volume of tooth 248 to increase the amount of detectable sample. In the illustrated embodiment, the volume of both the first tooth 248a and the second tooth 248b is 50 μL. In other embodiments, the volumes of the first tooth 248a and the second tooth 248b may be, for example, 40 μL, 45 μL, 55 μL, 60 μL, 65 μL, or any suitable volume. In the illustrated embodiment, the volume of each third tooth 248c is 123 μL. In other embodiments, the volume of each third tooth 248c may be, for example, 115 μL, 120 μL, 125 μL, 130 μL, 135 μL, or any suitable volume. The recommended maximum sample loading volume for the corresponding well should be less than the volume of tooth 248. For example, the recommended loading volume for a 123μL well is 100μL, and the recommended loading volume for a 50μL well is 35μL.

[0098] Continue to refer to Figure 9The distance between the first tooth 248a and the third end 236 of the middle portion 232 is a first distance 284. This first distance 284 separates the sample well formed by the first tooth 248a from the edge of the gel and the inner edge of the cavity 148 of the gel casting box 104. The first distance 284 can improve the quality, separation, and clarity of the bands during electrophoresis. This type of sample well positioning can reduce the probability of the bands formed in the gel exhibiting the following: the band ends curving upwards ("smile phenomenon"), the band ends curving downwards or the band edges curving downwards ("frowning phenomenon"), or otherwise deviating from the horizontal line to form a larger angle. The first distance 284 can be measured with reference to the first width 282a. For example, the first distance 284 can be equal to, greater than, or less than the first width 282a. The first distance 284 can be 1.5 times, 2 times, 2.5 times, etc., of the first width 282a, or less than this width. Alternatively, the first distance 284 can be measured directly numerically. For example, the first distance 284 can be 1 mm, 2 mm, 3 mm, 5 mm, 10 mm, 15 mm, 20 mm, etc. The first distance 284 can be any suitable distance that improves the quality, separation, and clarity of the electrophoretic bands. It should be understood that the distance between the second tooth 248b and the fourth end 240 of the intermediate portion 232 is the second distance. This second distance can be the same as the first distance 284. The second distance can have any of the characteristics described herein with reference to the first distance 284.

[0099] Continue to refer to Figure 9 The distance between the first end 188 of the elongated body 184 and the third end 236 of the intermediate portion 232 is a third distance 288. This third distance 288 can be adjusted to change the first distance 284. In some embodiments, the third distance 288 may be zero. It should be understood that the distance between the second end 192 of the elongated body 184 and the fourth end 240 of the intermediate portion 232 is a fourth distance. This fourth distance may be the same as the third distance 288. The fourth distance may have any of the characteristics described herein with reference to the third distance 288.

[0100] In some embodiments, by inserting the electrophoresis comb 108 described herein (e.g., an electrophoresis comb 108 with teeth 248 having at least one beveled surface) into the sample well formed by the gel material, a larger opening can be provided at the top of the well, making it easier to add samples using standard pipette tips of single-channel or multi-channel pipettes. Furthermore, the sample well described herein allows for the addition of larger quantities of sample into thinner gels (e.g., gels approximately 1 mm thick). The electrophoresis comb 108 can be used with the gel casting cartridge 104 described herein, or is suitable for any other suitable gel casting cartridge.

[0101] The sample loading wells formed in the gel by the 248 teeth described herein have a volume at least 12% larger than those formed by a standard electrophoresis comb. In some embodiments, the volume of the sample loading wells may be at least 10%, at least 20%, at least 50%, at least 100%, at least 150%, at least 175%, at least 200%, or at least 250% larger than those formed by a standard electrophoresis comb. Furthermore, the sample loading wells can increase the sample loading volume while maintaining or improving the clarity and separation of electrophoretic bands.

[0102] In some embodiments, by inserting an electrophoresis comb 108 with wedge-shaped teeth 248 (or wedge-shaped teeth 248) into a sample well (or wedge-shaped sample well) formed from the gel material, a larger opening is provided at the top of the well, making it easier to add samples using standard pipette tips from single-channel or multi-channel pipettes. Furthermore, the sample well including the wedge-shaped portion allows for the addition of a larger quantity of sample to a thinner gel (e.g., a gel approximately 1 mm thick). The electrophoresis comb 108 may be used with the gel casting cartridge 104 described herein, or may be suitable for any other suitable gel casting cartridge.

[0103] The wedge-shaped teeth 248 form a sample loading well in the gel, the volume of which can be at least 12% larger than that formed by a standard electrophoresis comb. In some embodiments, the volume of the sample loading well including the wedge-shaped portion can be at least 10%, at least 20%, at least 50%, at least 100%, at least 150%, at least 175%, at least 200%, or at least 250% larger than that formed by a standard electrophoresis comb. Furthermore, the sample loading well including the wedge-shaped portion can increase the sample loading volume while maintaining or improving the clarity and separation of electrophoretic bands.

[0104] In some embodiments, the electrophoretic comb 108 may be molded from any suitable plastic, including but not limited to the following polymers: for example, polyethylene terephthalate, polyvinyl chloride, polymethyl methacrylate, polystyrene, polyethylene, polymethyl polypropylene, cellulose acetate, copolymers, polycarbonate, or other suitable materials. In some embodiments, the dimensions of the teeth 248 may be consistent along the length of the electrophoretic comb 108 or vary along the length. The shape of the teeth 248 may be the same as or different from the wedge-shaped opening 156 of the adhesive cartridge 104.

[0105] refer to Figure 10 and 11 The electrophoresis comb 108 is coupled to the gel casting box 104. Edge 212 engages with the leading edge 172 of the abutment portion 152 on the fixing plate 116 to prevent gel moisture from escaping from the cavity 148. This design prevents the gel from drying out, for example, during storage or transportation of the gel electrophoresis assembly 100.

[0106] Figure 12 - Figure 15 Another embodiment of the electrophoresis comb 308 is shown. The electrophoresis comb 308 can replace the electrophoresis comb 108 in the gel electrophoresis assembly 100. Therefore, the electrophoresis comb 308 can be housed within the cavity 148 of the gel casting cartridge 104. For simplicity, this document only discusses the differences between the electrophoresis comb 308 and the electrophoresis comb 108. However, it should be understood that the electrophoresis comb 308 may include any of the features or functions described herein with reference to the electrophoresis comb 108. Similarly, the electrophoresis comb 108 may include any of the features or functions described herein with reference to the electrophoresis comb 308. Identical features and components of the electrophoresis comb 308 and the electrophoresis comb 108 are referred to similarly, but the reference number will be increased by 200 (or plus 200). For example, the electrophoresis comb 308 includes the same elongated body 384 as the elongated body 184 of the electrophoresis comb 108.

[0107] refer to Figure 12 - Figure 14 The electrophoresis comb 308 comprises a plurality of teeth 448. Each tooth 448 is identical and defines a width 482. The size of the tooth 448 affects its volume. The volume of the tooth 448 needs to be maximized, for example, to increase the amount of detectable sample. In the illustrated embodiment, the volume of each tooth 448 is 70 μL. In other embodiments, the volume of the tooth 448 may be, for example, 60 μL, 65 μL, 75 μL, 80 μL, 85 μL, or other suitable volumes. The recommended maximum sample loading volume for a corresponding sample well should be less than the volume of the tooth 448. For example, the recommended sample loading volume for a 70 μL sample well is 60 μL. The illustrated electrophoresis comb 308 comprises 20 teeth 448. In other embodiments, the electrophoresis comb 308 may comprise a different number of teeth 448 to achieve the desired number of sample wells.

[0108] Figure 16 - Figure 19 Another embodiment of the electrophoresis comb 508 is shown. Electrophoresis comb 508 can replace electrophoresis combs 108 and 308 in the gel electrophoresis assembly 100. Therefore, electrophoresis comb 508 can be housed in the cavity 148 of the gel casting cartridge 104. For simplicity, this document only discusses the differences between electrophoresis comb 508 and electrophoresis combs 108 and 308. However, it should be understood that electrophoresis comb 508 may include any of the features or functions described herein with reference to electrophoresis combs 108 and 308. Similarly, electrophoresis combs 108 and 308 may include any of the features or functions described herein with reference to electrophoresis comb 508. Identical features of electrophoresis comb 508 and electrophoresis comb 108 are designated similarly, but the reference number will be increased by 400 (or plus 400). For example, electrophoresis comb 508 includes an elongated body 584 identical to the elongated body 184 of electrophoresis comb 108.

[0109] refer to Figure 16 - Figure 18The electrophoresis comb 508 comprises a plurality of teeth 648. Each tooth 648 is identical and defines a width 682. The size of the tooth 648 affects its volume. The volume of the tooth 648 needs to be maximized, for example, to increase the amount of sample that can be detected. In the illustrated embodiment, the volume of each tooth 648 is 46 μL. In other embodiments, the volume of the tooth 648 may be, for example, 35 μL, 40 μL, 45 μL, 50 μL, 55 μL, 60 μL, or other suitable volumes. The recommended maximum sample loading volume for a corresponding well should be less than the volume of the tooth 648. For example, the recommended sample loading volume for a 46 μL well is 40 μL. The illustrated electrophoresis comb 508 comprises 26 teeth 648. In other embodiments, the electrophoresis comb 508 may comprise a different number of teeth 648 to achieve the desired number of wells.

[0110] method

[0111] Another embodiment described herein is a method for efficiently loading one or more large-volume samples into an electrophoresis gel, the method comprising: providing an assembly including the gel casting cartridge and at least one electrophoresis comb as described herein; preparing the gel by initiating a polymerization reaction of the electrophoresis gel pouring solution, pouring a sufficient amount of solution into the gel casting cartridge, inserting the electrophoresis comb into the gel casting cartridge, and waiting for the gel to polymerize; the gel can then be stored, packaged, or used immediately. When using the gel, the gel casting cartridge and electrophoresis comb assembly are assembled into an electrophoresis gel running apparatus. Running buffer (typically containing 25 mM Tris, 192 mM glycine, pH 8.3, and 0.1% sodium dodecyl sulfate (SDS)) is added, and the electrophoresis comb is removed from the assembly. The electrophoresis comb may be removed before adding the buffer. One or more samples are added to the sample wells, and an electric current is applied to begin electrophoresis.

[0112] Another embodiment described herein is a method for running multiple samples on an electrophoresis gel without causing band distortion in samples near the gel edge. The method includes: providing an assembly comprising the gel casting cartridge and at least one electrophoresis comb described herein; adding one or more samples to one or more wells; and applying an electric current for electrophoresis. The wells near the gel edge are recessed towards the center of the gel. This design prevents samples closest to the gel edge from being adversely affected by current conduction along the gel edge, resulting in band distortion or bending.

[0113] Another embodiment described herein is a method for preparing and loading electrophoresis gels while avoiding contamination of the sample wells by excess polymeric gel casting solution. The method includes: providing an assembly comprising a casting cartridge and at least one electrophoresis comb as described herein; filling the casting cartridge with sufficient electrophoresis gel casting solution, wherein the electrophoresis gel casting solution contains polyacrylamide, one or more buffer solutions, and optionally a detergent, and has been initiated for polymerization by adding a polymerization initiator and optionally a polymerization accelerator; inserting the electrophoresis comb into the casting cartridge and the unpolymerized gel; allowing the electrophoresis gel casting solution to polymerize for a period of time. The gel can then be stored, packaged, or used immediately. When using the gel, the casting cartridge and electrophoresis comb assembly are assembled into a running device. The electrophoresis comb is removed from the casting cartridge, wherein a minimal amount of excess polymeric gel casting solution remains in or around one or more sample wells after removal. Samples can be easily added to one or more sample wells.

[0114] Another embodiment described herein is a method for detecting low-abundance proteins or proteins in diluted samples by loading large sample volumes: providing an assembly comprising a gel casting cartridge and at least one electrophoresis comb as described herein; removing the electrophoresis comb from the assembly; adding one or more samples to one or more sample wells, wherein the volume of the sample wells is 40–100 μL; and applying an electric current for electrophoresis.

[0115] Reagent test kit

[0116] Another embodiment described herein is a kit comprising: an assembly containing the gel casting cartridge and at least one electrophoresis comb as described herein; a polymeric electrophoresis gel casting solution containing polyacrylamide, one or more buffer solutions and optional detergents; packaging material comprising a bag, pouch, plastic container or box; a buffer solution; and optional instructions for use.

[0117] Another embodiment is a ready-to-use pre-made electrophoresis gel assembly. This gel assembly includes a gel casting cartridge as described herein. The electrophoresis gel, containing polyacrylamide, buffer, and optional detergent, has been pre-cast using the electrophoresis comb described herein. The pre-made gel / cartridge / comb assembly is packaged in a sealed bag or pouch containing sufficient buffer or solution to keep the gel moist and prevent dehydration or drying. The cartridge is designed to avoid puncturing the sealed bag or pouch by eliminating sharp edges. One or more pre-made gel / cartridge / comb assemblies may be packaged together or individually. Multiple pre-made gel / cartridge / comb assemblies may be included in the kit. The kit may further include: SDS-PAGE concentrated run buffer, concentrated loading buffer, concentrated protein staining solution, concentrated destaining solution, molecular weight markers, containers, labels, and instructions for use. In some embodiments, the electrophoresis run device and / or power supply may be included in the kit. The kit may be supplied in a sealed box with labeling and branding.

[0118] It will be apparent to those skilled in the art that suitable modifications and adaptations can be made to the compositions, formulations, methods, processes, and applications described herein without departing from the scope of any embodiment or aspect thereof. The compositions and methods provided are exemplary and not intended to limit the scope of any specified embodiment. All the various embodiments, aspects, and options disclosed herein can be combined in any variation or iteration. The scope of the compositions, formulations, methods, and processes described herein includes all combinations of actual or potential embodiments, aspects, options, examples, and preferred embodiments described herein. Exemplary compositions and formulations described herein may omit any component, substitute for any component disclosed herein, or include any component disclosed elsewhere herein. In any composition or formulation disclosed herein, the mass ratio of each component to the mass of any other component in the formulation, or to the total mass of other components in the formulation, is deemed to be explicitly disclosed. If the meaning of any term in any patent or publication incorporated herein by reference conflicts with the meaning of the term used in this disclosure, the meaning of the term or phrase in this disclosure shall prevail. Furthermore, the foregoing discussion discloses and describes only exemplary embodiments. All patents and publications referenced herein are incorporated herein by reference for their specific teaching.

[0119] The various embodiments and aspects of the utility model described herein are summarized in the following clauses: Clause 1. An assembly for gel electrophoresis, comprising: A glue box comprising a fixed plate and a partition plate, the two coupled to form a cavity between the fixed plate and the partition plate; and An electrophoretic comb, comprising: A slender body having a first end and a second end; An intermediate portion connecting an elongated body and extending between a first end and a second end, the intermediate portion having a third end adjacent to the first end of the elongated body and a fourth end adjacent to the second end of the elongated body, the intermediate portion being received within a cavity; and Multiple teeth extending from the middle portion between the first and second ends, the multiple teeth being spaced apart from at least one of the third and fourth ends.

[0120] The assemblies described in Clause 2 and Clause 1, wherein: The plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width. The first tooth is spaced a first distance from the third end of the middle part, and The first distance is greater than the first width.

[0121] The assembly described in Clause 3, Clause 1, or 2, wherein: Multiple teeth include the second tooth closest to the fourth end of the middle portion. The second tooth has a second width. The second tooth is spaced a second distance from the fourth end of the middle portion, and The second distance is greater than the second width.

[0122] The assembly of any one of Clauses 4 and Clauses 1-3, wherein the first width is the same as the second width.

[0123] Assembly as described in any one of Clauses 5 and Clauses 1-4, wherein the first distance is the same as the second distance.

[0124] An assembly of any one of Clauses 6, 1-5, wherein each of the plurality of teeth has the same width.

[0125] The assembly described in any one of Clauses 7 and Clauses 1-6, wherein: Multiple teeth include a third tooth located between the first and second teeth. The third tooth has a third width, and The third width is greater than the first width and / or the second width.

[0126] The assembly of any one of Clauses 8 and 1-7, wherein the third width is greater than twice the first width and the third width is greater than twice the second width.

[0127] An assembly according to any one of Clauses 9 and 1-8, wherein each of the plurality of teeth comprises a first portion and a second portion located on the side of the first portion facing away from the intermediate portion, and the second portion is at an angle relative to the first portion.

[0128] The assembly of any one of Clauses 10 and Clauses 1-9, wherein the second part is at an angle of 15 to 25 degrees relative to the first part.

[0129] The assembly described in any one of Clauses 11 and Clauses 1-10, wherein: Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

[0130] The assembly of any one of Clauses 12, 1-11, wherein the volume of at least one of the plurality of teeth is at least 123 μL.

[0131] The assembly of any one of Clauses 13 and 1-12, wherein the volume of at least one of the plurality of teeth is at least 50 μL.

[0132] The assembly of any one of Clauses 14 and 1-13, wherein the volume of at least one of the plurality of teeth is at least 70 μL.

[0133] The assembly of any one of Clauses 15 and 1-14, wherein the volume of each of the plurality of teeth is at least 70 μL.

[0134] The assembly of any one of Clauses 16 and 1-15, wherein the volume of at least one of the plurality of teeth is at least 46 μL.

[0135] The assembly of any one of Clauses 17 and 1-16, wherein the volume of each of the plurality of teeth is at least 46 μL.

[0136] The assembly described in any one of Clauses 18 and Clauses 1-17, wherein: The volume of the first tooth is at least 50 μL. The volume of the second tooth is at least 50 μL, and The volume of the third tooth is at least 123 μL.

[0137] The assembly of any one of Clauses 19 and 1-18, wherein the plurality of teeth are spaced 1 to 5 mm from the third end and the plurality of teeth are spaced 1 to 5 mm from the fourth end.

[0138] The assembly of any one of Clauses 20 and 1-19, wherein the plurality of teeth are spaced 6 to 10 mm from the third end and the plurality of teeth are spaced 6 to 10 mm from the fourth end.

[0139] The assembly of any one of Clauses 21 and 1-20, wherein the plurality of teeth are spaced 11 to 15 mm from the third end and the plurality of teeth are spaced 11 to 15 mm from the fourth end.

[0140] Clause 22. A comb for gel electrophoresis, comprising: A slender body comprising a first end and a second end; A wall that extends at least partially along the outer periphery of a slender body; A middle portion connected to the wall and extending between the first and second ends of the elongated body, the middle portion having a third end and a fourth end opposite the third end, the third end adjacent to the first end of the elongated body and the fourth end adjacent to the second end of the elongated body; and Multiple teeth extending from the middle portion between the third and fourth ends, the multiple teeth being spaced apart from at least one of the third and fourth ends.

[0141] The electrophoretic combs described in Clauses 23 and 22, wherein: The plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width. The first tooth is spaced a first distance from the third end of the middle part, and The first distance is greater than the first width.

[0142] The electrophoretic comb described in Clause 24, Clause 22 or 23, wherein: Multiple teeth include the second tooth closest to the fourth end of the middle portion. The second tooth has a second width. The second tooth is spaced a second distance from the fourth end of the middle portion, and The second distance is greater than the second width.

[0143] The electrophoretic comb of any one of Clauses 25, 22-24, wherein the first width is the same as the second width.

[0144] The electrophoretic comb of any one of Clauses 26, 22-25, wherein the first distance is the same as the second distance.

[0145] An electrophoretic comb as described in any one of Clauses 27, 22-26, wherein each of the plurality of teeth has the same width.

[0146] The electrophoretic comb of any one of Clauses 28 and 22-27, wherein: Multiple teeth include a third tooth located between the first and second teeth. The third tooth has a third width, and The third width is greater than the first and second widths.

[0147] The electrophoretic comb of any one of Clauses 29 and 22-28, wherein the third width is greater than twice the first width and the third width is greater than twice the second width.

[0148] The electrophoretic comb of any one of Clauses 30 and 22-29, wherein each of the plurality of teeth comprises a first portion and a second portion located on the side of the first portion facing away from the middle portion, and the second portion is at an angle relative to the first portion.

[0149] The electrophoretic comb of any one of Clauses 31, 22-30, wherein the second part is at an angle of 15 to 25 degrees relative to the first part.

[0150] The electrophoretic comb described in any one of Clauses 32 and 22-31, wherein: Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

[0151] The electrophoretic comb of any one of Clauses 33, 22-31, wherein at least one of the plurality of teeth has a volume of at least 123 μL.

[0152] The electrophoretic comb of any one of Clauses 34 and 22-33, wherein at least one of the plurality of teeth has a volume of at least 50 μL.

[0153] The electrophoretic comb of any one of Clauses 35, 22-34, wherein at least one of the plurality of teeth has a volume of at least 70 μL.

[0154] The electrophoretic comb of any one of Clauses 36 and 22-35, wherein the volume of each of the plurality of teeth is at least 70 μL.

[0155] The electrophoretic comb of any one of Clauses 37, 22-36, wherein the volume of at least one of the plurality of teeth is at least 46 μL.

[0156] The electrophoretic comb of any one of Clauses 38 and 22-37, wherein the volume of each of the plurality of teeth is at least 46 μL.

[0157] The electrophoretic comb mentioned in any one of Clauses 39 and 22-38, wherein: The volume of the first tooth is at least 50 μL. The volume of the second tooth is at least 50 μL, and The volume of the third tooth is at least 123 μL.

[0158] The electrophoretic comb according to any one of Clauses 40 and 22-39, wherein the plurality of teeth are spaced 1 to 5 mm from the third end, and

[0159] The plurality of teeth are spaced 1 to 5 millimeters apart from the fourth end.

[0160] The electrophoretic comb of any one of Clauses 41, 22-40, wherein the plurality of teeth are spaced 6 to 10 mm from the third end and the plurality of teeth are spaced 6 to 10 mm from the fourth end.

[0161] The electrophoretic comb of any one of Clauses 42 and 22-41, wherein the plurality of teeth are spaced 11 to 15 mm from the third end and the plurality of teeth are spaced 11 to 15 mm from the fourth end.

[0162] Clause 43. An assembly for gel electrophoresis, comprising: A gel casting cartridge is configured to contain an electrophoresis gel casting solution. The cartridge includes a fixing plate and a partition plate coupled together to form a cavity between the fixing plate and the partition plate. The fixing plate has an upper edge adjacent to the cavity opening. An electrophoresis comb, comprising: A slender body that has: First end, The second end opposite to the first end, and The channel extending between the first end and the second end, Multiple teeth extending from the slender body between the first and second ends, and A wall extending at least partially along the outer periphery of an elongated body, a portion of which is configured to engage the upper edge of a fixing plate.

[0163] The assembly described in Clauses 44 and 43, wherein the wall extends from a first side of the elongated body and the channel extends along a second side of the elongated body opposite to the first side.

[0164] The assembly described in Clause 45, Clause 43 or 44, wherein the wall includes an edge extending between the first end and the second end and a lip extending between the first end and the second end, and the edge engages the upper edge of the fixing plate.

[0165] The assembly of any one of Clauses 46, 43-45, wherein a groove is formed between the edge and the lip, and the lip improves accessibility to the elongated body to assist in removing the electrophoretic comb from the gel casting box.

[0166] The assembly of any one of Clauses 47, 43-46, wherein the elongated body has a ramp near the edge within the groove, and the electrophoretic gel filling solution is configured to flow down the ramp and detach from the electrophoretic comb.

[0167] The assembly of any one of Clauses 48, 43-47, wherein the inclined surface forms a 30-degree angle with respect to the elongated body.

[0168] The assembly of any one of Clauses 49, 43-48, wherein the bevel is located on the side of the elongated body facing away from the channel.

[0169] The assembly described in any one of Clauses 50 and 43-49, wherein the fixing plate comprises: Fixed plate main body; An abutment portion extending along the main body of the fixing plate adjacent to the opening, the abutment portion having an upper edge and a tapered edge; and A guide rail extending along the main body of the fixing plate and spaced apart from the abutment portion has a tapered edge.

[0170] The assembly described in any one of Clauses 51 and 43-50, wherein: The electrophoretic comb comprises a middle section located between a slender body and multiple teeth. The middle section extends between the first and second ends of the slender body. The middle portion has a third end and a fourth end opposite to the third end, and The third and fourth ends connect to the inner edge of the cavity.

[0171] The assembly of any one of Clauses 52, 43-51, wherein the channel extends beyond the third and fourth ends.

[0172] An assembly as described in any one of Clauses 53 and 43-52, wherein the plurality of teeth are spaced apart from at least one of the third and fourth ends.

[0173] The assembly described in any one of Clauses 54 and 43-53, wherein the assembly further comprises a polymerized electrophoretic gel casting solution, The solution includes polyacrylamide, one or more buffer solutions, and optionally a detergent.

[0174] Clause 55. A comb for gel electrophoresis, comprising: The slender body includes: First end; The second end opposite the first end; and A channel extending between the first end and the second end; A wall, at least partially extending along the outer periphery of an elongated body, the wall including a flange extending between a first end and a second end, and a lip extending between the first end and the second end; and Multiple teeth extending from the elongated body between the first and second ends.

[0175] The electrophoretic comb described in Clauses 56 and 55, wherein the wall extends from a first side of the elongated body and the channel extends along a second side of the elongated body away from the first side.

[0176] The electrophoretic comb described in Clause 57, Clause 55 or 56, wherein a groove is formed between the edge and the lip, and the lip enhances accessibility to the slender body.

[0177] The electrophoretic comb of any one of Clauses 58, 55-57, wherein the elongated body has a bevel near the edge of the groove, and the electrophoretic gel filling solution is configured to flow down the bevel and detach from the electrophoretic comb.

[0178] The electrophoretic comb of any one of Clauses 59, 55-58, wherein the bevel is at a 30-degree angle to the elongated body.

[0179] The electrophoretic comb of any one of Clauses 60 and 55-59, wherein the bevel is located on the side of the elongated body opposite to the channel.

[0180] The electrophoretic comb as described in any one of Clauses 61 and 55-60, wherein: The electrophoretic comb comprises a middle section located between a slender body and multiple teeth. The middle section extends between the first and second ends of the slender body, and The middle part has a third end and a fourth end opposite to the third end.

[0181] The electrophoretic comb of any one of Clauses 62 and 55-61, wherein the channel extends beyond the third and fourth ends.

[0182] The electrophoretic comb of any one of Clauses 63 and 55-62, wherein the plurality of teeth are spaced apart from at least one of the third and fourth ends.

[0183] Clause 64. An assembly for gel electrophoresis, comprising: A glue box includes a fixed plate and a partition plate, which are coupled to form a cavity between the fixed plate and the partition plate. The fixed plate has an upper edge adjacent to the opening of the cavity; and An electrophoretic comb, containing An elongated body having a first end and a second end opposite to the first end, and A wall, at least partially extending along the outer periphery of an elongated body, includes a flange extending between a first end and a second end, the flange contacting an edge of a fixing plate, and Multiple teeth extending from the wall between the first and second ends of the slender body.

[0184] The assembly described in Clauses 65 and 64, wherein the elongated body includes a channel extending between a first end and a second end.

[0185] The assembly described in Clause 66, Clause 64 or 65, wherein the wall extends from a first side of the elongated body and the channel extends along a second side of the elongated body away from the first side.

[0186] An assembly according to any one of Clauses 67, 64-66, wherein each of the plurality of teeth comprises a first portion and a second portion located on the side of the first portion facing away from the wall, and the second portion is at an angle relative to the first portion.

[0187] An assembly as described in any one of Clauses 68, 64-67, wherein the second part forms an angle of 15 to 25 degrees relative to the first part.

[0188] An assembly as described in any one of Clauses 69, 64-68, wherein: Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

[0189] An assembly according to any one of Clauses 70, 64-69, wherein the volume of at least one of the plurality of teeth is at least 123 μL.

[0190] An assembly according to any one of Clauses 71, 64-70, wherein the volume of at least one of the plurality of teeth is at least 50 μL.

[0191] An assembly according to any one of Clauses 72, 64-71, wherein the volume of at least one of the plurality of teeth is at least 70 μL.

[0192] The assembly of any one of Clauses 73, 64-72, wherein the volume of each of the plurality of teeth is at least 70 μL.

[0193] An assembly according to any one of Clauses 74 and 64-73, wherein the volume of at least one of the plurality of teeth is at least 46 μL.

[0194] An assembly according to any one of Clauses 75, 64-74, wherein the volume of each of the plurality of teeth is at least 46 μL.

[0195] Clause 76. An electrophoresis comb for gel electrophoresis, configured to be mounted in a gel casting cartridge including an upper edge, the electrophoresis comb comprising: A slender body comprising a first end and a second end; A wall, at least partially extending along the outer periphery of an elongated body, includes a flange extending between a first end and a second end, the flange being configured to engage with the upper edge of a groove.

[0196] The intermediate portion connecting the wall and extending between the first and second ends of the elongated body; and

[0197] Multiple teeth extending from the middle section.

[0198] The electrophoretic comb described in Clauses 77 and 76, wherein the elongated body contains a channel extending between a first end and a second end.

[0199] The electrophoretic comb described in Clause 78, Clause 76 or 77, wherein the wall extends from a first side of the elongated body and the channel extends along a second side of the elongated body away from the first side.

[0200] The electrophoretic comb of any one of Clauses 79, 76-78, wherein each of the plurality of teeth comprises a first portion and a second portion located on the side of the first portion facing away from the middle portion, and the second portion is at an angle relative to the first portion.

[0201] The electrophoretic comb of any one of Clauses 80, 76-79, wherein the second part is at an angle of 15 to 25 degrees relative to the first part.

[0202] The electrophoretic comb mentioned in any one of Clauses 81 and 76-80, wherein: Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

[0203] The electrophoretic comb of any one of Clauses 82, 76-81, wherein at least one of the plurality of teeth has a volume of at least 123 μL.

[0204] The electrophoretic comb of any one of Clauses 83, 76-82, wherein at least one of the plurality of teeth has a volume of at least 50 μL.

[0205] The electrophoretic comb of any one of Clauses 84, 76-83, wherein at least one of the plurality of teeth has a volume of at least 70 μL.

[0206] The electrophoretic comb of any one of Clauses 85, 76-84, wherein the volume of each of the plurality of teeth is at least 70 μL.

[0207] The electrophoretic comb of any one of Clauses 86, 76-85, wherein at least one of the plurality of teeth has a volume of at least 46 μL.

[0208] The electrophoretic comb of any one of Clauses 87, 76-86, wherein the volume of each of the plurality of teeth is at least 46 μL.

[0209] Clause 88. A reagent kit comprising: Assembly as described in any one of Clauses 1-21, 43-54, or 64-75; A polymeric electrophoresis gel preparation solution consisting of polyacrylamide, one or more buffer solutions, and optionally a detergent; Packaging consisting of bags, pouches, plastic containers, or boxes; Buffer solutions; and Optional instruction manual.

[0210] Clause 89. A method for efficiently loading one or more large-volume samples into an electrophoresis gel, the method comprising: Provide assemblies as described in any one of Clauses 1-21, 43-54 or 64-75; Remove the electrophoresis comb from the assembly; and One or more samples are loaded into one or more sample wells.

[0211] Clause 90. A method for running multiple samples on an electrophoresis gel and avoiding sample band distortion near the gel edge, the method comprising: Provide assemblies as described in any one of Clauses 1-21, 43-54 or 64-75; Remove the electrophoresis comb from the assembly; Load one or more samples into one or more sample wells; and Electrophoresis is performed by applying an electric current.

[0212] Clause 91. A method for preparing and loading an electrophoresis gel while avoiding contamination of at least one sample loading well within the polymer gel in the gel casting cartridge by excessive polymer gel preparation solution, the method comprising: Provide assemblies as described in any one of Clauses 1-21, 43-54 or 64-75; Add sufficient electrophoresis gel casting solution to fill the gel casting box, wherein the electrophoresis gel casting solution contains polyacrylamide, One or more buffer solutions and optional detergents, and the polymerization reaction of the electrophoretic gel casting solution has been initiated by adding a polymerization initiator and optional polymerization accelerator; Insert the electrophoretic comb as described in any of Clauses 22-42, 55-63 or 76-87 into the gel casting box; Allow the electrophoretic gel casting solution to polymerize for a period of time; Remove the electrophoresis comb from the gel casting box, ensuring that one or more sample wells contain a minimal amount of excess polymerized gel casting solution; and One or more samples are loaded into one or more sample wells.

[0213] Clause 92, A method for detecting low-abundance proteins or proteins in diluted samples by loading large sample volumes: Provide assemblies as described in any one of Clauses 1-21, 43-54 or 64-75; Remove the electrophoresis comb from the assembly; One or more samples are introduced into one or more wells, the wells having a volume of 40–100 μL; and Electrophoresis is performed by applying an electric current.

[0214] Clause 93, use the assembly described in any one of Clauses 1-21, 43-54 or 64-75 to perform electrophoresis on the sample.

[0215] The uses described in Clauses 94 and 93, wherein the sample is a low-abundance protein or a protein in a diluted sample.

[0216] Clause 95, use the electrophoresis comb described in any one of Clauses 22-42, 55-63 or 76-87 to fill electrophoresis gel, wherein the volume of the sample well formed by the electrophoresis comb is about 40-100 μL.

Claims

1. An assembly for gel electrophoresis, characterized in that, include: The glue box includes a fixed plate and a partition plate, which are coupled together to form a cavity between the fixed plate and the partition plate; as well as Electrophoretic comb, including: A slender body having a first end and a second end; An intermediate portion connecting an elongated body and extending between a first end and a second end, the intermediate portion having a third end adjacent to the first end of the elongated body and a fourth end adjacent to the second end of the elongated body, the intermediate portion being received within a cavity; and Multiple teeth extending from the middle portion between the first and second ends, the multiple teeth being spaced apart from at least one of the third and fourth ends.

2. The assembly according to claim 1, characterized in that, The plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width. The first tooth is spaced a first distance from the third end of the middle part, and The first distance is greater than the first width.

3. The assembly according to claim 2, characterized in that, Multiple teeth include the second tooth closest to the fourth end of the middle portion. The second tooth has a second width. The second tooth is spaced a second distance from the fourth end of the middle portion, and The second distance is greater than the second width.

4. The assembly according to claim 3, characterized in that, The first width is the same as the second width.

5. The assembly according to claim 3, characterized in that, The first distance is the same as the second distance.

6. The assembly according to claim 1, characterized in that, Each tooth in the multiple teeth has the same width.

7. The assembly according to claim 3, characterized in that, Multiple teeth include a third tooth located between the first and second teeth. The third tooth has a third width, and The third width is greater than the first width and / or the second width.

8. The assembly according to claim 7, characterized in that, The third width is greater than twice the first width, and the third width is greater than twice the second width.

9. The assembly according to claim 1, characterized in that, Each of the multiple teeth includes a first portion and a second portion located on the side of the first portion facing away from the middle portion, with the second portion forming an angle relative to the first portion.

10. The assembly according to claim 9, characterized in that, The second part forms an angle of 15 to 25 degrees relative to the first part.

11. The assembly according to claim 9, characterized in that, Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

12. The assembly according to claim 1, characterized in that, At least one of the teeth has a volume of at least 123 μL.

13. The assembly according to claim 12, characterized in that, At least one of the teeth has a volume of at least 50 μL.

14. The assembly according to claim 1, characterized in that, At least one of the teeth has a volume of at least 70 μL.

15. The assembly according to claim 14, characterized in that, The volume of each tooth in the plurality of teeth is at least 70 μL.

16. The assembly according to claim 1, characterized in that, At least one of the teeth has a volume of at least 46 μL.

17. The assembly according to claim 16, characterized in that, The volume of each tooth in the plurality of teeth is at least 46 μL.

18. The assembly according to claim 7, characterized in that, The volume of the first tooth is at least 50 μL. The volume of the second tooth is at least 50 μL, and The volume of the third tooth is at least 123 μL.

19. The assembly according to claim 1, characterized in that, Multiple teeth are spaced 1 to 5 mm from the third end, and multiple teeth are spaced 1 to 5 mm from the fourth end.

20. The assembly according to claim 1, characterized in that, Multiple teeth are spaced 6 to 10 mm from the third end, and multiple teeth are spaced 6 to 10 mm from the fourth end.

21. The assembly according to claim 1, characterized in that, Multiple teeth are spaced 11 to 15 mm from the third end, and multiple teeth are spaced 11 to 15 mm from the fourth end.

22. An electrophoretic comb for gel electrophoresis, characterized in that, include: A slender body comprising a first end and a second end; A wall that extends at least partially along the outer periphery of a slender body; A middle portion connected to the wall and extending between the first and second ends of the elongated body, the middle portion having a third end and a fourth end opposite to the third end, the third end being adjacent to the first end of the elongated body and the fourth end being adjacent to the second end of the elongated body; as well as Multiple teeth extending from the middle portion between the third and fourth ends, the multiple teeth being spaced apart from at least one of the third and fourth ends.

23. The electrophoretic comb according to claim 22, characterized in that, The plurality of teeth includes a first tooth closest to the third end of the middle portion, the first tooth having a first width. The first tooth is spaced a first distance from the third end of the middle part, and The first distance is greater than the first width.

24. The electrophoretic comb according to claim 23, characterized in that, Multiple teeth include the second tooth closest to the fourth end of the middle portion. The second tooth has a second width. The second tooth is spaced a second distance from the fourth end of the middle portion, and The second distance is greater than the second width.

25. The electrophoretic comb according to claim 24, characterized in that, The first width is the same as the second width.

26. The electrophoretic comb according to claim 24, characterized in that, The first distance is the same as the second distance.

27. The electrophoretic comb according to claim 22, characterized in that, Each tooth in the multiple teeth has the same width.

28. The electrophoretic comb according to claim 24, characterized in that, Multiple teeth include a third tooth located between the first and second teeth. The third tooth has a third width, and The third width is greater than the first and second widths.

29. The electrophoretic comb according to claim 28, characterized in that, The third width is greater than twice the first width, and the third width is greater than twice the second width.

30. The electrophoretic comb according to claim 22, characterized in that, Each of the multiple teeth includes a first portion and a second portion located on the side of the first portion facing away from the middle portion, with the second portion forming an angle relative to the first portion.

31. The electrophoretic comb according to claim 30, characterized in that, The second part forms an angle of 15 to 25 degrees relative to the first part.

32. The electrophoretic comb according to claim 30, characterized in that, Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

33. The electrophoretic comb according to claim 22, characterized in that, At least one of the teeth has a volume of at least 123 μL.

34. The electrophoretic comb according to claim 33, characterized in that, At least one of the teeth has a volume of at least 50 μL.

35. The electrophoretic comb according to claim 22, characterized in that, At least one of the teeth has a volume of at least 70 μL.

36. The electrophoretic comb according to claim 35, characterized in that, The volume of each tooth in the plurality of teeth is at least 70 μL.

37. The electrophoretic comb according to claim 22, characterized in that, At least one of the teeth has a volume of at least 46 μL.

38. The electrophoretic comb according to claim 37, characterized in that, The volume of each tooth in the plurality of teeth is at least 46 μL.

39. The electrophoretic comb according to claim 28, characterized in that, The volume of the first tooth is at least 50 μL. The volume of the second tooth is at least 50 μL, and The volume of the third tooth is at least 123 μL.

40. The electrophoretic comb according to claim 22, characterized in that, Multiple teeth are spaced 1 to 5 mm from the third end, and multiple teeth are spaced 1 to 5 mm from the fourth end.

41. The electrophoretic comb according to claim 22, characterized in that, Multiple teeth are spaced 6 to 10 mm from the third end, and multiple teeth are spaced 6 to 10 mm from the fourth end.

42. The electrophoretic comb according to claim 22, characterized in that, Multiple teeth are spaced 11 to 15 mm from the third end, and multiple teeth are spaced 11 to 15 mm from the fourth end.

43. An assembly for gel electrophoresis, characterized in that, include: A gel casting box is configured to contain an electrophoresis gel casting solution. The gel casting box includes a fixed plate and a partition plate, which are coupled to form a cavity between the fixed plate and the partition plate. The fixed plate has an upper edge adjacent to the opening of the cavity. as well as Electrophoretic comb, including: The slender body has the following characteristics: First end; The second end opposite the first end; and A channel extending between the first end and the second end; Multiple teeth extending from the slender body between the first and second ends; and A wall extending at least partially along the outer periphery of an elongated body, a portion of which is configured to engage the upper edge of a fixing plate.

44. The assembly according to claim 43, characterized in that, The wall extends from the first side of the elongated body, and the channel extends along the second side of the elongated body away from the first side.

45. The assembly according to claim 43, characterized in that, The wall includes a flange extending between a first end and a second end and a lip extending between the first end and the second end, and the flange engages with the upper edge of the fixing plate.

46. ​​The assembly according to claim 45, characterized in that, A groove is formed between the flange and the lip, and the lip improves accessibility to the slender body to help remove the electrophoretic comb from the gel casting box.

47. The assembly according to claim 46, characterized in that, The slender body has a bevel near the edge of the groove, and the electrophoretic gel filling solution is configured to flow down the bevel and detach from the electrophoretic comb.

48. The assembly according to claim 47, characterized in that, The inclined surface forms a 30-degree angle with respect to the slender body.

49. The assembly according to claim 47, characterized in that, The inclined surface is located on the side of the slender body facing away from the channel.

50. The assembly according to claim 43, characterized in that, The fixing plate includes: Fixed plate body, An abutment portion extending along the main body of the fixing plate adjacent to the opening, the abutment portion having an upper edge and a tapered edge, and A guide rail extending along the main body of the fixing plate and spaced apart from the abutment portion has a tapered edge.

51. The assembly according to claim 43, characterized in that, The electrophoretic comb comprises a middle section located between a slender body and multiple teeth. The middle section extends between the first and second ends of the slender body. The middle portion has a third end and a fourth end opposite to the third end, and The third and fourth ends connect to the inner edge of the cavity.

52. The assembly according to claim 51, characterized in that, The channel extends beyond the third and fourth ends.

53. The assembly according to claim 51, characterized in that, Multiple teeth are spaced apart from at least one of the third and fourth ends.

54. An electrophoretic comb for gel electrophoresis, characterized in that, include: The slender body includes: First end; The second end, opposite to the first end; as well as A channel extending between the first end and the second end; A wall, at least partially extending along the periphery of an elongated body, the wall including a flange extending between a first end and a second end, and a lip extending between the first end and the second end; as well as Multiple teeth extending from the elongated body between the first and second ends.

55. The electrophoretic comb according to claim 54, characterized in that, The wall extends from the first side of the elongated body, and the channel extends along the second side of the elongated body away from the first side.

56. The electrophoretic comb according to claim 54, characterized in that, A groove is formed between the flange and the lip, and the lip improves accessibility to the slender body.

57. The electrophoretic comb according to claim 56, characterized in that, The slender body has a bevel near the edge of the groove, and the electrophoretic gel filling solution is configured to flow down the bevel and detach from the electrophoretic comb.

58. The electrophoretic comb according to claim 57, characterized in that, The inclined surface forms a 30-degree angle with respect to the slender body.

59. The electrophoretic comb according to claim 57, characterized in that, The inclined surface is located on the side of the slender body facing away from the channel.

60. The electrophoretic comb according to claim 54, characterized in that, The electrophoretic comb comprises a middle section located between a slender body and multiple teeth. The middle section extends between the first and second ends of the slender body, and The middle part has a third end and a fourth end opposite to the third end.

61. The electrophoretic comb according to claim 60, characterized in that, The channel extends beyond the third and fourth ends.

62. The electrophoretic comb according to claim 60, characterized in that, Multiple teeth are spaced apart from at least one of the third and fourth ends.

63. An assembly for gel electrophoresis, characterized in that, include: A glue box includes a fixed plate and a partition plate, which are coupled to form a cavity between the fixed plate and the partition plate. The fixed plate has an upper edge adjacent to the opening of the cavity. as well as Electrophoretic comb, including: An elongated body having a first end and a second end opposite to the first end; A wall, at least partially extending along the outer periphery of an elongated body, the wall including a flange extending between a first end and a second end, the flange contacting an edge of a fixed plate; as well as Multiple teeth extending from the wall between the first and second ends of the slender body.

64. The assembly according to claim 63, characterized in that, The elongated body includes a channel extending between the first end and the second end.

65. The assembly according to claim 64, characterized in that, The wall extends from the first side of the elongated body, and the channel extends along the second side of the elongated body opposite to the first side.

66. The assembly according to claim 63, characterized in that, Each of the multiple teeth includes a first portion and a second portion located on the side of the first portion facing away from the wall, with the second portion forming an angle with respect to the first portion.

67. The assembly according to claim 66, characterized in that, The second part forms an angle of 15 to 25 degrees relative to the first part.

68. The assembly according to claim 66, characterized in that, Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

69. The assembly according to claim 63, characterized in that, At least one of the teeth has a volume of at least 123 μL.

70. The assembly according to claim 69, characterized in that, At least one of the teeth has a volume of at least 50 μL.

71. The assembly according to claim 63, characterized in that, At least one of the teeth has a volume of at least 70 μL.

72. The assembly according to claim 71, characterized in that, The volume of each tooth in the plurality of teeth is at least 70 μL.

73. The assembly according to claim 63, characterized in that, At least one of the teeth has a volume of at least 46 μL.

74. The assembly according to claim 73, characterized in that, The volume of each tooth in the plurality of teeth is at least 46 μL.

75. An electrophoresis comb for gel electrophoresis, configured to be mounted in a gel casting cartridge including an upper edge, characterized in that, The electrophoretic comb includes: A slender body comprising a first end and a second end; A wall, at least partially extending along the outer periphery of an elongated body, the wall including a flange extending between a first end and a second end, the flange being configured to engage with the upper edge of a groove. The intermediate portion connecting the wall and extending between the first and second ends of the elongated body; and Multiple teeth extending from the middle section.

76. The electrophoretic comb according to claim 75, characterized in that, The elongated body includes a channel extending between the first end and the second end.

77. The electrophoretic comb according to claim 76, characterized in that, The wall extends from the first side of the elongated body, and the channel extends along the second side of the elongated body away from the first side.

78. The electrophoretic comb according to claim 75, characterized in that, Each of the multiple teeth includes a first portion and a second portion located on the side of the first portion facing away from the middle portion, with the second portion forming an angle relative to the first portion.

79. The electrophoretic comb according to claim 78, characterized in that, The second part forms an angle of 15 to 25 degrees relative to the first part.

80. The electrophoretic comb according to claim 75, characterized in that, Each of the multiple teeth includes a third part located on the side of the second part opposite to the first part. The first part defines the first plane. The third part defines the third plane, and The first plane is parallel to the third plane.

81. The electrophoretic comb according to claim 75, characterized in that, At least one of the teeth has a volume of at least 123 μL.

82. The electrophoretic comb according to claim 81, characterized in that, At least one of the teeth has a volume of at least 50 μL.

83. The electrophoretic comb according to claim 75, characterized in that, At least one of the teeth has a volume of at least 70 μL.

84. The electrophoretic comb according to claim 83, characterized in that, The volume of each tooth in the multiple teeth is at least 70 μL.

85. The electrophoretic comb according to claim 75, characterized in that, At least one of the teeth has a volume of at least 46 μL.

86. The electrophoretic comb according to claim 85, characterized in that, The volume of each tooth in the plurality of teeth is at least 46 μL.

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