JP5299102B2 - Gel cassette and manufacturing method thereof - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a gel cassette for easily and certainly forming a linear gel interface. <P>SOLUTION: The gel cassette 1 includes a first substrate 10, a second substrate bonded to the first substrate to form a gel filling section between it and the first substrate, a first gel section formed on a first end side of the gel filling section, a second gel section formed on a second end side of the gel filling section to provide an interface with the first gel section along a predetermined boundary line P, a first filling opening 16 formed at a position on the first end side of the gel filling section than the boundary line, of at least one of the first substrate and the second substrate, to communicate with the gel filling section, and a second filling opening 17 and deaeration opening 18 formed at a position on the second end side of the gel filling section from the boundary line, of at least one of the first substrate and the second substrate, to communicate with the gel filling section. <P>COPYRIGHT: (C)2011,JPO&amp;INPIT

Description

  The present invention relates to a gel cassette, and more particularly to a gel cassette used for protein electrophoresis and a method for producing the same.

  Conventionally, two-dimensional electrophoresis of proteins is known as one of the techniques for profiling proteins. Proteins have different properties in terms of charge and molecular weight. Therefore, rather than separating individual proteins from the proteome, which is a mixture of many proteins, depending on only the charge or molecular weight, two-dimensional electrophoresis combining both proteins can be used to achieve higher protein resolution. There is an advantage that it can be separated.

In two-dimensional electrophoresis, the sample is applied to the first dimension gel and subjected to isoelectric focusing, then the first dimension gel is taken out and applied to the second dimension gel, and the second dimension separation is performed based on the molecular weight. I do.
In this case, a gel cassette called a second-dimensional gel chip (second separation unit) in which a second-dimensional gel layer is disposed between two substrates is often used (see, for example, Patent Document 1).

  There are several constitutional patterns of the second-dimensional gel layer, but those having two types of gel layers of a separation gel and a concentrated gel are common. In the second-dimensional gel chip having the second-dimensional gel layer having such a configuration, it is important to keep the interface between the separation gel and the concentrated gel (gel interface) in a straight line in order to maintain high measurement accuracy. .

As a method for forming a gel interface, two methods, a two-step method and a one-step method, are mainly known. In the two-step method, after the gel precursor solution (sol) of one gel layer is filled in the space formed between the substrates, water is overlaid, and the upper surface of the layer made of the gel is straightened by the water layer. In this method, water is removed and the gel precursor solution of the other gel layer is filled.
The one-step method is a method in which glycerin or the like is mixed in one gel precursor solution to increase the specific gravity, and is filled at the same time or almost simultaneously with the other gel precursor solution to form a linear interface due to the difference in specific gravity between the two. .

JP 2007-64848 A

However, since the water removal process is required in the two-step method, the work of forming the gel interface becomes complicated, and as a result, neither the flatness of the gel interface of the second-dimensional gel chip nor the production efficiency is sufficient. There is.
Further, in the one-step method, after filling one gel precursor solution, the other gel precursor solution is filled at a short interval, and therefore two types of gel precursor solutions may be excessively mixed in the filling process. As a result, there is a problem that separation by specific gravity becomes insufficient and a linear interface may not be formed.

  This invention is made | formed in view of the said situation, and it aims at providing the gel cassette which can form a gel interface linearly more easily and reliably, and its manufacturing method.

The gel cassette according to the first aspect of the present invention is a gel cassette that constitutes a second-dimensional gel chip that performs two-dimensional electrophoresis by placing a first-dimensional gel chip after performing first-dimensional electrophoresis. , a first substrate, a second substrate which the gel-filled portion is bonded to the first substrate so as to form between the first substrate, formed on the first end side of the gel filling portion A first gel part, a second gel part formed on the second end side of the gel filling part so as to have an interface with the first gel part along a predetermined boundary line; At least one of the substrate and the second substrate, a first filling port formed to communicate with the gel filling portion at a position closer to the first end portion than the boundary line, the first substrate, and the first substrate At least one of the second substrates, the gel at a position closer to the second end than the boundary line A second filling port formed so as to communicate with the filling portion, and at least one of the first substrate and the second substrate at a position closer to the second end side than the boundary line, to the gel filling portion A slit for inserting the first-dimensional gel chip when in use, wherein the first substrate communicates with the gel filling portion, a part of the first substrate extends in parallel with the boundary line. A part of the opening of the slit has a protrusion that can be attached to and detached from the opening, and a first sealing member that fills the protrusion in the opening and seals the opening is provided. At least one of the one filling port, the second filling port, and the deaeration port is formed by an opening of a portion of the slit that is not sealed by the first sealing member. .

At least one of the first substrate and the second substrate has a through hole communicating with the gel filling portion, and at least one of the first filling port, the second filling port, and the degassing port is: An elastically deformable second sealing member may be attached and formed so as to close the through hole.

In the method for producing a gel cassette according to the second aspect of the present invention, a gel in which a first gel portion made of a first gel and a second gel portion made of a second gel form an interface along a predetermined boundary line. have a layer, a process for the preparation of gel cassettes constituting the second dimension Geruchippu performing two-dimensional electrophoresis by placing the first dimension Geruchippu after the first dimensional electrophoresis, partially the A first substrate that extends parallel to the boundary line and has a slit into which the first-dimensional gel chip is inserted in use ; and a second substrate that forms a gel-filled portion between the first substrate and the first substrate and the first substrate At least one of the second substrates is formed to have an opening including the slit, and the first substrate and the first substrate are formed so that a gel filling portion is formed between the first substrate and the second substrate . a bonding step of bonding together the second substrate, said opening The gel filling portion is attached to a position closer to the first end portion than the boundary line by an opening of a portion of the opening that is not sealed by the sealing member. A first filling port formed so as to communicate with the first filling port, a second filling port formed so as to communicate with the gel filling unit at a position closer to the second end than the boundary line, and the boundary line A degassing port formed so as to communicate with the gel filling unit at a position closer to the second end side, the first filling port, the second filling port, and the degassing port At least one of the sealing step formed by opening the portion of the slit that is not sealed by the sealing member, and the first end side of the gel-filled portion with respect to the first end side of the gel filling portion . from the filling port, the filling precursor solution of the first gel to the gel filling portion 1 And Le filling step, said from said second filling port of the second end portion side of the gel-filled portion of the boundary line, the precursor solution of the first gel precursor liquid lighter in specific gravity than the second gel gel And a second gel filling step for filling the filling portion.

  According to the gel cassette and the manufacturing method thereof of the present invention, it is possible to provide a gel cassette in which a linear gel interface is formed more easily and reliably.

It is a perspective view which shows the gel cassette of one Embodiment of this invention in a partial cross section. It is a top view of the same gel cassette before gel layer formation. It is sectional drawing in the AA of FIG. It is a flowchart which shows the flow of the manufacturing method of the gel cassette. It is a figure which shows operation | movement of the 1st gel filling process of the manufacturing method. It is a figure which shows operation | movement of the 1st filling port sealing process of the manufacturing method. It is a figure which shows operation | movement of the 2nd gel filling process of the manufacturing method. It is a figure which shows operation | movement of the 2nd filling port sealing process of the manufacturing method.

Hereinafter, a gel cassette according to an embodiment of the present invention will be described with reference to FIGS.
FIG. 1 is a perspective view showing the gel cassette 1 of the present embodiment in a partial cross section, FIG. 2 is a plan view of the gel cassette 1 before forming a gel layer (described later), and FIG. 3 is taken along the line AA in FIG. It is sectional drawing.

  The gel cassette 1 of the present embodiment is a second-dimensional gel chip used for two-dimensional electrophoresis of proteins. As shown in FIGS. 1 to 3, the first substrate 10 and the second substrate 10 made of a transparent resin are used. The substrate 20 and the substrate 20 are integrally joined.

The first substrate 10 includes a rectangular bottom plate 11 (including a substantially rectangular shape) and a wall surface 12 rising on the first surface 11A from the periphery of the bottom plate 11. Buffer tanks 14A and 14B are formed by partition walls 13 on the first surface 11A of the first end 10A of the first substrate 10 and the second end 10B opposite to the first end 10A, respectively.

  As shown in FIGS. 1 and 3, slits 15 </ b> A and 15 </ b> B extending in the longitudinal direction of the buffer tank 14 </ b> B are formed in a portion of the bottom plate 11 that forms the bottom surface of the buffer tank 14 </ b> B. The slits 15A and 15B penetrate the bottom plate 11 in the thickness direction, and the first-dimensional gel chip is inserted into the slit 15B near the partition wall 13 when the gel cassette 1 is used. The slit 15A near the second end portion 10B is used for bringing the buffer filled in the buffer tank 14B into contact with a gel layer 31 described later when the gel cassette 1 is used.

  One slit 15C having the same shape as the slits 15A and 15B is also formed in a portion of the bottom plate 11 that forms the bottom surface of the buffer tank 14A. The slit 15C is used to bring the buffer filled in the buffer tank 14A into contact with the gel layer 31 when the gel cassette 1 is used.

  The 2nd board | substrate 20 has the structure substantially the same as the 1st board | substrate 10, and has the wall surface 22 which stood | started up from the periphery of the baseplate 21 to the 1st surface 21A side as shown in FIG. Thereby, the recessed part 23 is formed in the 1st surface 21A side.

  The first substrate 10 and the second substrate 20 are integrally formed by bonding, welding, or the like with the second surface 11B opposite to the first surface 11A of the bottom plate 11 and the first surface 21A of the bottom plate 21 facing each other. It is joined. Thereby, the gel filling part 30 which is a space for filling the gel is formed between the first substrate 10 and the second substrate 20. In the present embodiment, the second surface 11B of the bottom plate 11 is flat and the gel filling portion 30 is formed solely by the recess 23, but instead the second surface 11B has a recess. The gel filling portion may be formed by integrating the recess and the recess 23.

As shown in FIG. 1, a gel layer 31 filled with a gel is formed in the gel filling portion 30. The gel layer 31 is made of a separation gel (first gel), and the end of the gel filling unit 30 on the first end 10A side of the first substrate 10 (hereinafter, this end is referred to as the first end of the gel filling unit 30). It is defined as a portion 30A.) An end portion (hereinafter referred to as the second end portion 10B side) of the first substrate 10 of the gel filling portion 30, which is composed of a first gel portion 32 and a concentrated gel (second gel) located closer to each other. The end portion is defined as a second end portion 30B of the gel filling portion 30.). The boundary line P formed by the interface between the first gel portion 32 and the second gel portion 33 is formed between the partition walls 13 and at a desired position near the second end portion 10B. That is, the interface between the first gel portion 32 and the second gel portion 33 is formed along the boundary line P.
The boundary line P is at least parallel to (including substantially parallel to) the slit 15B into which the first-dimensional gel chip is inserted. In the gel cassette 1 of the present embodiment, all three slits 15A, 15B, and 15C are the boundary line P. And parallel (including substantially parallel).

The separation gel is a gel for separating proteins according to their molecular weight during electrophoresis, and the concentrated gel moves the protein on the first-dimensional chip toward the interface between the first gel portion 32 and the second gel portion 33. It is a gel that concentrates in the direction of movement while letting go. As for any gel, various known gels including a general polyacrylamide gel can be appropriately selected and employed.
In addition, the gel layer 31 of this embodiment is filled by the 1 step method, and the above-mentioned interface is formed. The detailed procedure of gel filling will be described later.

  As shown in FIG. 1, a sealing member 35 is attached to the slit 15C. The sealing member 35 seals the slit 15 </ b> C from the manufacturing of the gel cassette 1 to the time of use, leaving a part, and prevents the gel layer 31 from drying, such as an elastic member such as rubber, a seal-like member, etc. Can be suitably employed. In the present embodiment, elastic urethane rubber is employed as the sealing member 35. The sealing member 35 has a protrusion 35A extending in the longitudinal direction of the sealing member 35, and the protrusion 35A is detachably attached to the first substrate 10 so as to fill the slit 15C.

As shown in FIG. 2, an opening 35 </ b> B communicating with the slit 15 </ b> C is formed at one end in the longitudinal direction of the sealing member 35. Thereby, the opening 35B and a part of the slit 15C exposed without being sealed by the sealing member 35 by overlapping with the opening 35B are integrated with each other to open on the upper surface of the sealing member 35, and to fill the gel. A first filling port 16 communicating with the portion 30 is formed.
The shape in the vicinity of the end of the slit 15 </ b> C that becomes a part of the first filling port 16 may be appropriately set in consideration of the dimensions of a nozzle used for gel filling described later. For example, according to the diameter of the nozzle, the width dimension in the vicinity of the end of the slit 15C is set to be the same (including substantially the same) as the diameter or width dimension of the opening 35B, or is completely the same as the shape of the opening 35B in plan view. May be set to the same (including substantially the same). In the present embodiment, the end of the slit 15A that overlaps the opening 35B is expanded to a length that is substantially the same as the diameter of the opening 35A.

A sealing member 36 having substantially the same material and configuration as the sealing member 35 is attached to the slits 15A and 15B. The sealing member 36 has protrusions 36A and 36B having the same shape as the protrusion 35A, and the protrusions 36A and 36B are attached to and detached from the first substrate 10 so as to fill the slits 15A and 15B, respectively, leaving both ends in the longitudinal direction. Can be attached freely.
Openings 36 </ b> C and 36 </ b> D communicating with the slits 15 </ b> A and 15 </ b> B are formed at both ends in the longitudinal direction of the sealing member 36. As a result, the opening 36C and the region near one end of the slits 15A and 15B that are exposed without being overlapped with the opening 36C are integrated into an opening on the upper surface of the sealing member 36. A second filling port 17 communicating with the filling unit 30 is formed. Then, the opening 36D and the area near the other end of the slits 15A and 15B that are exposed without being overlapped with the opening 36D are integrated into an opening on the upper surface of the sealing member 36, and the gel is filled. A deaeration port 18 communicating with the portion 30 is formed.

Similar to the first filling port 16 described above, the shape in the vicinity of the ends of the slits 15A and 15B, which are part of the second filling port 17, is also set in consideration of the dimensions of the nozzles used for gel filling described later. Specifically, the end portions of the two slits 15A and 15B are combined into one on the second filling port 17 side, and the width dimension is set to be substantially the same as the diameter of the opening 36C.
Regarding the deaeration port 18, the ends of the slits 15A and 15B are not combined into one, and the respective ends are exposed overlapping the opening 36D. As will be described later, the deaeration port 18 passes only when the air in the gel filling portion 30 escapes to the outside of the gel cassette 1 when the gel precursor solution is filled, so that the ends of the slits 15A and 15B overlapping the opening 36D. Although it is not necessary to set the shape like the second filling port 17 to the part, there is no problem even if the same shape setting as the second filling port 17 is performed.

In the present embodiment, the openings 35B and 36C forming part of the first filling port 16 and the second filling port 17 are circular, and the opening 36D forming a part of the deaeration port 18 has a substantially rectangular shape. Although an example is shown, the shape of the opening formed in the sealing members 35 and 36 is not particularly limited, and when considering the above-described nozzle dimensions, the diameter and the like are determined according to the specific shape of the opening. The maximum width dimension may be set as appropriate.
Moreover, when the gel cassette 1 is completed, as shown in FIG. 1, the first filling port 16, the filling second port 17, and the deaeration port 18 are all closed with a sealing material 37 (desorption). Refer to FIG. This will be described in detail in the manufacturing method of the gel cassette 1 below.

The manufacturing method of the gel cassette 1 of this embodiment comprised as mentioned above is demonstrated.
FIG. 4 is a flowchart showing a flow of a manufacturing method of the gel cassette 1. The manufacturing method includes a joining step S10 for joining the first substrate 10 and the second substrate 20, a first gel filling step S20 for filling the separated gel precursor solution into the gel filling part 30 from the first filling port 16, and A first filling port sealing step S30 for sealing the first filling port 16 after filling the separated gel precursor solution, and a second gel filling step S40 for filling the gel filling unit 30 from the second filling port 17 with the concentrated gel precursor solution. And a second filling port sealing step S50 for sealing the second filling port 17 and the deaeration port 18 after filling the concentrated gel precursor solution.

First, in the bonding step of step S10, the first substrate 10 and the second substrate 20 are bonded together by bonding, welding, or the like with the second surface 11B and the first surface 21A facing each other. Thereby, the gel filling part 30 which can be filled with a gel precursor liquid between the 1st board | substrate 10 and the 2nd board | substrate 20 is formed.
Next, the sealing members 35 and 36 are attached to the first substrate 10. Accordingly, the three slits 15A, 15B, and 15C are sealed except for the first filling port 16, the second filling port 17, and the deaeration port 18.

Subsequently, the gel layer 31 is formed in the gel filling portion 30 by a one-step method.
In the first gel filling step of step S <b> 20, the gel filling unit 30 is held so that the second filling port 17 and the deaeration port 18 are positioned relatively higher than the first filling port 16. Then, as shown in FIG. 5, the nozzle 41 for supplying the separated gel precursor solution 32 </ b> A is inserted into the first filling port 16, and the separated gel precursor solution 32 </ b> A is filled into the gel filling unit 30. Glycerin, sucrose, or the like is added to the separation gel precursor solution 32A so that the interface between the first gel portion 31 and the second gel portion 32 is favorably formed, and the specific gravity with the concentrated gel precursor solution described later. It is preferable to increase the difference.

As the gel filling part 30 is filled with the separated gel precursor solution 32A, the air in the gel filling part 30 flows from the second filling port 17 and the degassing port 18 to the outside of the gel cassette 1 as indicated by an arrow A in FIG. Run away.
The posture of the gel filling unit 30 in the first gel filling step S20 is particularly limited as long as the relative height relationship between the second filling port 17 and the degassing port 18 and the first filling port 17 is satisfied as described above. However, as shown in FIG. 5, it is preferable that the gel filling portion 30 be vertical with the first end portion 10A side of the first substrate 10 on which the first filling port 16 is formed facing down. If it does in this way, since it should just be filled with separation gel precursor liquid 32A to boundary line P which forms the interface of the 1st gel part 32 and the 2nd gel part 33, adjustment of the amount of filling can be performed appropriately.
When the separation gel precursor solution 32A is filled in the gel filling unit 30 by a predetermined amount, the supply of the separation gel precursor solution 32A from the nozzle 41 is stopped, and the process proceeds to the first filling port sealing step of step S30.

In the first filling port sealing step S <b> 30, the nozzle 41 is pulled out from the first filling port 17 and the first filling port 16 is sealed by the sealing material 37. The sealing method is not particularly limited, and various methods such as adhesion of a sheet coated with a hot melt adhesive can be used in addition to the application of the sealing material 37 as in the present embodiment. When using a sealing material in which release paper is bonded to the adhesive layer, a part of the adhesive layer is adhered onto the sealing member 35 and the sealing material is temporarily fixed to the sealing member, and the nozzle 41 is removed. The first filling port 16 may be sealed with the sealing material after removing the release paper.
After the 1st filling port 16 is sealed as shown in FIG. 6, a process progresses to the 2nd gel filling process of step S40.

In the second gel filling step S40, as shown in FIG. 7, the nozzle 42 for supplying the concentrated gel precursor solution is inserted into the second filling port 17, and the concentrated gel precursor solution 33A is filled into the gel filling unit 30. At this time, the concentrated gel precursor solution 33A is at a lower speed than the first gel filling step S20 so as not to disturb the upper surface of the separated gel precursor solution 31A serving as the interface between the first gel portion 32 and the second gel portion 33 excessively. Filling is preferred. In this step, the air in the gel filling unit 30 escapes from the degassing port 18 to the outside of the gel cassette 1 as indicated by an arrow A in FIG.
When the concentrated gel precursor solution 33A is filled in the predetermined amount of gel filling unit 30 set in advance, the supply of the separated gel precursor solution 33A from the nozzle 42 is stopped, and the process is the second filling port sealing step of step S50. Proceed to

In the second filling port sealing step S <b> 50, the nozzle 42 is removed from the second filling port 17, and the second filling port 17 and the deaeration port 18 are sealed with the sealing material 37 in the same manner as the first filling port 16. . However, this is an example, and at least one of the second filling port 17 and the deaeration port 18 may be sealed by a method different from that of the first filling port 16.
After the second filling port 17 and the deaeration port 18 are sealed, when the gel filling part 30 is left to stand vertically, the separated gel precursor solution 32A and the concentrated gel precursor solution 33A each have a boundary line due to the difference in specific gravity between them. A good linear interface along the boundary line P is formed by separating the lower side and the upper side of P. When left still for a predetermined time, moisture evaporates from the separated gel precursor solution 32A and the concentrated gel precursor solution 33A, and each precursor solution becomes a first gel portion 32 and a second gel portion 33, respectively, and a gel cassette provided with a gel layer 31. 1 is completed. Thus, a series of manufacturing steps is completed.

  When the gel cassette 1 manufactured as described above is used, the gel cassette 1 is left standing with the first surface 11A facing upward, and the sealing members 35 and 36 are removed to expose the slits 15A, 15B, and 15C. A first-dimensional gel chip (not shown) is inserted into the slit 15B, the first-dimensional gel is applied to the second gel portion 33, the buffer is filled in the buffer tanks 14A, 14B, and the buffer and the gel layer 31 are brought into contact with each other. Then, protein electrophoresis can be performed by passing an electrode through the buffers in the buffer layers 14A and 14B and energizing them.

  As explained above, in the manufacturing method of the gel cassette 1 of the present embodiment, in the first gel filling step S20, the boundary line P where the interface between the first gel part 32 and the second gel part 33 is formed is formed. From the first filling port 16 located near the first end 30 </ b> A of the gel filling unit 30, a separation gel precursor solution 32 </ b> A that later constitutes the first gel unit 32 is filled. Thereafter, in the second gel filling step S40, the concentrated gel precursor solution 33A that will later constitute the second gel portion 33 from the second filling port 17 located closer to the second end 30B of the gel filling portion 30 than the boundary line P. Is filled.

Accordingly, when the separation gel precursor solution 32A is filled, the separation gel precursor solution 32A does not touch the inner wall surface of the gel filling portion 30 closer to the second end of the gel filling portion 30 than the boundary line P, and is concentrated. When the gel precursor solution 33A is filled, it does not come into contact with the separated gel precursor solution 32A attached to the inner wall surface.
As a result, the separation of the separated gel precursor solution 32A and the concentrated gel precursor solution 33A is suitably performed, and the gel layer 31 having a good interface between the first gel portion 32 and the second gel portion 33 is reliably obtained by a one-step method. And it can manufacture efficiently.

  Further, in the gel cassette 1 of the present embodiment, the first filling port 16, the first filling port 16, the first filling port 16, the first filling port 16, Since the two filling ports 17 and the deaeration port 18 are formed, it is not necessary to provide the filling port or the like in the first substrate 10 or the second substrate 10 separately from the slits, and the structure is simple and efficient to manufacture. Can do.

  Furthermore, since the sealing members 35 and 36 that form part of the first filling port 16, the second filling port 17, and the deaeration port 18 are formed of an elastic material, they are the same as the opening 35B or the like. By filling the gel precursor solution using a nozzle 41 or the like having a slightly larger diameter, the first filling port 16 or the second filling port 17 and the nozzles 41 and 42 can be connected in a watertight manner. As a result, leakage of the gel precursor solution during filling can be suitably prevented.

  Although one embodiment of the present invention has been described above, the technical scope of the present invention is not limited to the above embodiment, and various modifications can be made without departing from the spirit of the present invention. is there.

For example, in the gel cassette 1 of the present embodiment, the first filling port 16, the second filling port 17, and the deaeration port 18 are made using a part of the slits 15 </ b> A to 15 </ b> C that overlap with the opening provided in the sealing member. (Hereinafter, these three are collectively referred to as “first filling port 16 etc.”), but instead of this, it is overlapped with the opening and communicated with the gel filling unit 30. A through hole may be formed in the first substrate 10 apart from the slit 15C, and the first filling port 16 and the like may be formed by integrating the through hole and the opening 35A.
Further, both ends of the slit 15A and the like that remain without being sealed by sealing the region excluding both ends of the slit 15A and the like using a sealing member that has no opening and whose longitudinal dimension is shorter than the slit 15A and the like. The first filling port 16 or the like may be formed using the above. If it does in this way, the 1st filling port 16 grade | etc., Can be formed more easily with the sealing member of a simple shape.
Further, by combining the above-described aspects, at least one of the first filling port 16 and the like may be formed so as to use a part of the slit 15A and the like.

  Moreover, the formation positions of the first filling port 16 and the like are not limited to the vicinity of the first end portion 30A and the second end portion 30B of the gel filling portion 30 as in the present embodiment. That is, the first filling port 16 is closer to the first end portion 30A than the boundary line P where the interface between the first gel portion 32 and the second gel portion 33 is formed, and the second end portion 30B side is closer to the boundary line P. The second filling port 17 and the deaeration port 18 may be formed respectively. Accordingly, the first filling port 16 and the like may be formed on the second substrate 20 as long as they communicate with the gel filling unit 30, or may be formed on the wall surface 22 instead of the bottom plate 21.

  Furthermore, it is not necessary that all the first filling ports 16 and the like are formed on the same substrate, and at least one may be formed on a substrate different from the other two.

Furthermore, in the present embodiment, the example in which the first filling port 16 and the like are sealed with the sealing material 37 after filling the gel precursor solution has been described. Instead, the first substrate 10 or the second substrate 20 is provided. The first filling port 16 and the like may be formed by attaching an elastically deformable sealing member to the through hole that is communicated with the gel filling unit 30 so as to close the through hole.
In such a case, a hollow needle-like nozzle having a certain rigidity is pierced into the sealing member to fill the gel precursor solution, and after completion of filling, the nozzle is pulled out of the sealing member. The formed hole is automatically closed by the elastic force of the sealing member. Therefore, not only can the nozzle be connected to the gel filling portion in a watertight manner, but also the first filling port and the like are sealed at the same time that the nozzle is pulled out, so the first filling port sealing step S20 and the second filling port sealing step S40. Etc. can be omitted to simplify the manufacturing process.

  Further, in the present embodiment, the example in which the separation gel is first filled with the separation gel as the first gel has been described. Since the separation gel is usually higher in concentration than the concentrated gel, the one-step method is easier to manufacture if the separation gel precursor solution is filled first. This is because the separation gel is always the first gel. Does not mean. Therefore, if the specific gravity of the concentrated gel can be made relatively higher than that of the separation gel by adding glycerin or the like, the concentrated gel precursor solution may be filled first with the concentrated gel as the first gel.

DESCRIPTION OF SYMBOLS 1 Gel cassette 10 1st board | substrate 15A, 15B, 15C Slit 16 1st filling port 17 2nd filling port 18 Deaeration port 20 2nd board | substrate 30 Gel filling part 30A 1st end part 30B 2nd end part 31 Gel layer 32 1st 1 gel part 32A separation gel precursor solution (first gel precursor solution)
33 Second gel part 33A Concentrated gel precursor solution (precursor solution of second gel)
35, 36 Sealing member P Boundary line S10 Joining step S20 First gel filling step S40 Second gel filling step

Claims (3)

A gel cassette constituting a second-dimensional gel chip for performing two-dimensional electrophoresis by arranging a first-dimensional gel chip after performing the first-dimensional electrophoresis,
A first substrate;
A second substrate joined to the first substrate so as to form a gel filling portion with the first substrate;
A first gel portion formed on the first end side of the gel filling portion;
A second gel portion formed on the second end side of the gel filling portion so as to have an interface with the first gel portion along a predetermined boundary line;
A first filling port formed at least one of the first substrate and the second substrate at a position closer to the first end portion than the boundary line so as to communicate with the gel filling portion;
At least one of the first substrate and the second substrate at a position closer to the second end than the boundary line, and a second filling port formed to communicate with the gel filling unit;
A deaeration port formed at least one of the first substrate and the second substrate at a position closer to the second end side than the boundary line so as to communicate with the gel filling unit;
Equipped with a,
The first substrate is
Communicating with the gel filling part, a part extends parallel to the boundary line, and has a slit into which the first-dimensional gel chip is inserted during use,
The opening of a part of the slit is
A first sealing member that has a detachable protrusion in the opening and seals the opening by filling the protrusion in the opening;
At least one of the first filling port, the second filling port, and the degassing port is:
The gel cassette, wherein the gel cassette is formed by an opening of a portion of the slit that is not sealed by the first sealing member . At least one of the first substrate and the second substrate has a through hole communicating with the gel filling portion,
At least one of the first filling port, the second filling port, and the deaeration port is formed by attaching an elastically deformable second sealing member so as to close the through hole. The gel cassette according to claim 1, wherein A second gel portion comprising a first gel portion and a second gel comprising a first gel, have a gel layer to form an interface along a predetermined boundary line, after the first dimensional electrophoresis A method for producing a gel cassette constituting a second-dimensional gel chip for performing two-dimensional electrophoresis by arranging a first-dimensional gel chip ,
Part extends parallel to the boundary line, a first substrate having a slit inserting the first dimension Geruchippu in use, and a second substrate to form a gel filling portion between said first substrate, said first At least one of the one substrate and the second substrate is formed to have an opening including the slit,
As the gel filling portion is formed between the first substrate the second substrate, a bonding step of bonding together the said first substrate and the second substrate,
A sealing member that seals a part of the opening is attached, and the gel is placed at a position closer to the first end side than the boundary line by an opening of the opening that is not sealed by the sealing member. A first filling port formed so as to communicate with the filling unit, a second filling port formed so as to communicate with the gel filling unit at a position closer to the second end than the boundary line; A deaeration port formed to communicate with the gel filling part at a position on the second end side from the boundary line; and
At least one of the first filling port, the second filling port, and the deaeration port is formed by a sealing step of the slit that is not sealed by the sealing member;
A first gel filling step of filling the gel filling portion with the first gel precursor solution from the first filling port on the first end side of the gel filling portion with respect to the boundary line;
From the second filling port on the second end side of the gel filling part with respect to the boundary line, the gel filling part is filled with the second gel precursor liquid having a specific gravity lighter than that of the first gel precursor liquid. A second gel filling step;
A method for producing a gel cassette, comprising:

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