JP5057225B2 - Electrophoresis support holder and electrophoresis chip - Google Patents

Description

  The present invention relates to an electrophoresis support holder and an electrophoresis chip. More specifically, the present invention relates to an electrophoresis support holder and an electricity provided with an accommodating portion for accommodating an electrophoresis support therein. The present invention relates to an electrophoresis chip.

  Electrophoresis technology that separates biopolymer samples such as DNA, RNA, and proteins extracted from animal and plant tissues and cells into electrophoresis gels based on differences in size, form, etc. is very important in the life science field. Technology. In particular, when separating proteins by electrophoresis, two-dimensional electrophoresis combining SDS-polyacrylamide gel electrophoresis based on differences in protein molecular weight and isoelectric focusing based on differences in isoelectric points is widely used. It has been. Information such as the size, form, and charge of the sample can be obtained by electrophoresis, and the sample can be collected from the gel after electrophoresis and subjected to further analysis tests.

  Usually, electrophoresis gel is stored in a gel container (gel cassette) and used as a chip. The gel set is composed of a substrate having a gel container and a lid formed so as to cover the substrate, and the lid is detachably attached to the substrate. Conventionally, such a gel cassette made of glass has been widely used, but in recent years, a gel cassette made of synthetic resin has been used. Synthetic resins are suitable for mass production because they are easy to mold and have the advantage that they are not easily damaged. On the other hand, the gel cassette made of synthetic resin has a problem that it is difficult to form a gel in the gel container and the gel is easily peeled off. In addition, there is a problem that the separation pattern of the sample separated in the electrophoresis gel tends to be disturbed.

The structure for solving the problem in such a synthetic resin gel cassette is disclosed in Patent Documents 1 to 3. Patent Document 1 discloses a configuration in which a gel cassette is coated with a resin containing an ethylenically unsaturated group, and an electrophoretic gel is copolymerized with the coating resin. This prevents the electrophoresis gel from peeling off from the gel cassette. The configuration described in Patent Document 2 coats the gel cassette with silicon oxide and prevents the migration of oxygen from the gel cassette to the gel, thereby enabling uniform gel production and good electrophoresis. The configuration described in Patent Document 3 coats the gel cassette with vinylidene chloride and prevents oxygen migration from the gel cassette to the gel, thereby enabling uniform gel production and good electrophoresis.
US Pat. No. 4,415,428 (published November 15, 1983) US Pat. No. 5,685,967 (published November 11, 1997) International publication WO 90/13020 pamphlet (published on November 1, 1990)

  In order to further analyze the sample after electrophoresis, processing such as sampling from the electrophoresis gel, peeling and drying of the electrophoresis gel, and storage are performed. Therefore, it is necessary that the electrophoresis gel being sampled is firmly held by the gel cassette and that the electrophoresis gel can be easily peeled off from the gel cassette. In the configuration described in Patent Document 1, since the electrophoresis gel is firmly bonded to the gel cassette, the electrophoresis gel is sufficiently held by the gel cassette, but the electrophoresis gel cannot be peeled from the gel cassette. In the configuration described in Patent Document 2, it is possible to peel the electrophoresis gel from the gel cassette. However, when the electrophoresis gel is exposed by removing the lid of the gel cassette for sampling, the exposure time elapses. Therefore, the electrophoresis gel is dried and peeled off from the gel cassette and bent. In the configuration described in Patent Document 3, no measures are taken to prevent gel peeling.

  The present invention has been made in view of the above problems, and an object of the present invention is to provide an electrophoresis support holder that can sufficiently hold an electrophoresis gel and can peel the electrophoresis gel. There is.

  An electrophoretic support holder according to the present invention is an electrophoretic support holder that accommodates an electrophoretic support therein and includes a housing portion having a detachable lid. Therefore, it is characterized in that a resin layer containing a water-soluble resin is provided on at least a part of the surface of the housing portion that contacts the support for electrophoresis.

  In addition, in order to solve the above problems, an electrophoresis chip according to the present invention includes the electrophoresis support holder and an electrophoresis support housed inside the electrophoresis support holder. It is characterized by that.

  Thereby, the support for electrophoresis can be suitably held. Specifically, for example, even after removing the lid of the container for sampling after electrophoresis and exposing the support for electrophoresis, the support for electrophoresis does not peel from the container, so that sampling is easily performed. It is possible. On the other hand, when the electrophoresis support is peeled off for analysis, storage, etc., the electrophoresis support can be easily peeled off. In addition, it is possible to obtain a good separation pattern of the sample by electrophoresis without disturbing the electrophoresis. Furthermore, since it is not an organic solvent but a layer of water-soluble resin, it can be suitably used for a housing portion made of synthetic resin.

  Moreover, in the support holder for electrophoresis according to the present invention, the water-soluble resin is a water-soluble polymer derived from synthesis such as polyvinyl alcohol, polyacrylic acid, polymethacrylic acid, polyvinylamine, polyallylamine, starch, cellulose. It is preferably selected from the group consisting of dielectrics, pectin, alginic acid, polysaccharides such as agarose and pullulan, and proteins such as gelatin. As a result, the support for electrophoresis can be sufficiently brought into close contact with the accommodating portion, so that sampling after electrophoresis is easy and the support for electrophoresis can be easily peeled off.

  Further, in the electrophoresis support holder according to the present invention, the resin layer is formed on the outer peripheral portion of the bottom portion of the housing portion, and is at least 5 of the area of the surface in contact with the electrophoresis support body of the housing portion. It preferably has an area of ~ 100%. Thereby, it is possible to hold | maintain the support body for electrophoresis fully.

  In the electrophoresis support holder according to the present invention, it is preferable that the resin layer is formed on the entire inner surface of the bottom portion of the housing portion. Thereby, since the area which the resin layer and the support body for electrophoresis contact is large, the support body for electrophoresis can fully be hold | maintained. Further, the resin layer can be easily formed.

  Further, in the electrophoresis support holder according to the present invention, it is preferable that the housing portion is made of a synthetic resin. Thereby, the accommodating part can be easily formed. In addition, since it is harder to break than the glass container, damage to the electrophoresis support due to breakage of the container or the like can be avoided when the container is attached to or detached from the electrophoresis apparatus.

  Furthermore, in the electrophoresis support holder according to the present invention, it is preferable that the surface of the housing portion that contacts the electrophoresis support is subjected to a hydrophilic treatment. Thereby, it is possible to form a resin layer easily.

  According to the electrophoretic support holder of the present invention, the housing portion for accommodating the electrophoretic support has a resin layer containing a water-soluble resin on at least a part of the surface in contact with the electrophoretic support. Therefore, it is possible to suitably hold the support for electrophoresis.

  An embodiment of the present invention will be described below with reference to the drawings.

  FIG. 1 is a schematic view showing a gel cassette (electrophoretic support holder) 1 according to the present invention. As shown in FIG. 1, the gel cassette 1 includes a gel container (container) 2 composed of a base 2a and a lid 2b, and serves as a container for holding an electrophoresis gel (electrophoresis support). Used. The gel container 2 holds the electrophoresis gel in the gel holding region 4, and the resin layer 3 is provided on a part of the surface (the surface on the gel holding region 4 side) of the gel container 2 that contacts the electrophoresis gel. It has been.

  The base material portion 2a and the lid portion 2b constituting the gel containing portion 2 are preferably made of a synthetic resin such as a styrene resin, a styrene-acrylonitrile copolymer, an acrylic resin, a polyester resin, a cellulose resin, or a vinyl chloride resin. However, it is not limited to these. If the gel accommodating part 2 is formed of a synthetic resin, it can be easily formed by an injection molding method or the like. Moreover, since the synthetic resin gel container 2 is less likely to be damaged than the glass gel container, the gel container 2 may be damaged when the gel cassette 1 is attached to or detached from the electrophoresis apparatus. Electrophoretic gel damage can be avoided.

  The base material portion 2a is formed to have a concave depression, and this depression becomes a gel holding region 4 that holds the electrophoresis gel. The lid portion 2b is a plate-like body and is provided so as to cover and close the gel holding region 4. The lid portion 2b is detachable from the base material portion 2a. After the electrophoresis gel is accommodated in the gel holding region 4 of the base material portion 2a, the lid portion 2b is attached to the base material portion 2a and subjected to electrophoresis. After the electrophoresis, the lid 2b is removed and sampling is performed. Although the attachment method of the cover part 2b with respect to the base material part 2a is not specifically limited, It can attach by ultrasonic welding etc.

  The resin layer 3 provided in the gel container 2 is a thin layer containing a water-soluble resin as a main component. The resin layer 3 is formed by a known thin layer forming means, and can be formed by, for example, applying a water-soluble resin solution using a spin coater and coating the gel container 2 with the water-soluble resin. It is not limited to. The gel cassette 1 can suitably hold the electrophoresis gel by providing the resin layer 3 on the surface of the gel container 2 that is in contact with the electrophoresis gel. In addition, it is possible to obtain a good separation pattern by electrophoresis without hindering electrophoresis. Furthermore, since it is a layer of water-soluble resin, not an organic solvent, it can be suitably used for the gel housing part 2 made of synthetic resin.

  In FIG. 1, the resin layer 3 is formed on the entire inner surface of the bottom of the gel container 2, but the resin layer 3 is formed on a part of the inner surface of the bottom of the gel container 2, that is, on the outer periphery of the bottom. May be. Moreover, the resin layer 3 may be formed on the surface (inner surface of the side portion) that contacts the side end portion of the electrophoresis gel of the gel containing portion 2. Specifically, it is preferable that the resin layer 3 has an area of at least 5 to 100% of the area of the surface of the gel container 2 that contacts the electrophoresis gel. Thereby, the electrophoresis gel can be sufficiently held in the gel holding region 4.

  The water-soluble resin that is the main component of the resin layer 3 realizes a configuration in which the electrophoresis gel is sufficiently held and the electrophoresis gel and the gel container 2 are in close contact with each other so that the gel can be peeled off. This water-soluble resin is preferably one that does not dissolve completely in water and absorbs water to swell and become a gel state, such as polyvinyl alcohol (PVA), polyacrylic acid, polymethacrylic acid, polyvinylamine, polyallylamine, etc. Preferably selected from the group consisting of synthetic water-soluble polymers, starches, cellulose dielectrics, pectin, alginic acid, agarose, pullulan and other polysaccharides, and proteins such as gelatin, but are not limited thereto. Among these, polyvinyl alcohol is particularly capable of forming a film having low oxygen permeability, and in particular, a radical polymer is used to form a support for electrophoresis using a polyacrylamide material in which oxygen is an obstacle to polymerization. It is effective in the case.

  The degree of close contact between the gel container 2 and the electrophoresis gel that are in close contact with each other through the resin layer 3 is sufficient to remove the lid 2b after electrophoresis to expose the electrophoretic gel and perform sampling or the like. To the extent that the electrophoresis gel can be retained. The sampling time varies depending on the amount of sample, the purpose of sampling, etc., but it may take about 1 to 2 hours. If sampling is not performed carefully, the gel will dry and shrink and deform and lose the desired spot position. There is a case. Since the gel container 2 and the electrophoresis gel are in close contact with each other by a force stronger than the force by which the electrophoresis gel shrinks due to drying during the sampling time, sampling can be performed easily. Further, when the electrophoresis gel is peeled off from the gel container 2 for analysis, storage, etc., the electrophoresis gel can be easily peeled off.

  The water-soluble resin is preferably a completely saponified resin. Thereby, peeling of the electrophoresis gel when the electrophoresis gel is exposed can be suppressed for a longer time. The fully saponified resin is intended to be a resin that is almost completely saponified, and is intended to be a resin having a higher degree of saponification than a partially saponified resin that contains a non-saponified portion at a certain ratio. Saponification is a reaction in which an ester is alkali-hydrolyzed, and the degree of saponification indicates the ratio of saponification.

  Furthermore, the polymerization degree of the water-soluble resin is preferably 2000 to 4000. Thereby, peeling of the electrophoresis gel when the electrophoresis gel is exposed can be suppressed for a longer time. If the degree of polymerization is less than 2000, the time during which peeling of the electrophoresis gel can be suppressed is shortened, which is not preferable.

  When the synthetic resin gel container 2 is used, the surface of the gel container 2 on which the resin layer 3 is formed (the surface in contact with the electrophoresis gel) is highly hydrophobic, and the resin layer 3 mainly contains a water-soluble resin. Is difficult to form. Therefore, it is preferable that the surface of the gel container 2 on which the resin layer 3 is formed is subjected to a hydrophilic treatment. The hydrophilic treatment can be performed by a known hydrophilic means, and the method is not particularly limited. In particular, it is preferable to hydrophilize by coating a silicon oxide film, because the adhesion of the gel container 2 and the silicon oxide film, and the silicon oxide film and the resin layer 3 are increased. Moreover, since the oxygen permeability of the silicon oxide film is relatively low, the migration of oxygen from the synthetic resin gel container 2 is suppressed, and there is an effect that the polymerization of the electrophoresis gel solution is not inhibited.

  If the gel cassette 1 according to the present invention is used, it is possible to sufficiently hold the electrophoresis gel until the sample is electrophoresed and sampled from the gel after the electrophoresis. It is possible to peel off.

  The electrophoresis chip according to the present invention contains an electrophoresis gel in the gel holding region 4 of the gel cassette 1 and is sealed to form a single part. The sample is attached to the electrophoresis apparatus and separated by electrophoresis. Used for applications. The electrophoresis chip only needs to include at least an electrophoresis gel in the gel cassette 1, but may further include components (buffers, etc.) necessary for electrophoresis. Further, a sample to be separated by electrophoresis may be provided.

  A sample to be separated using the electrophoresis chip according to the present invention is a substance to be analyzed by electrophoresis, and is obtained from a biological material (for example, a living organism, a body fluid, a cell line, a tissue culture, or a tissue fragment). These preparations can be used preferably. In particular, protein samples, DNA samples, and RNA samples can be suitably used as samples. These samples may be prestained with a fluorescent sample or the like (fluorescent staining or fluorescent labeling).

  The electrophoresis gel accommodated in the gel holding region 4 is separated on the basis of the difference in the size, form, etc. of the sample by electrophoresis in the electrophoresis gel, and may be simply referred to as a gel. Conventionally generally used gel materials such as polyacrylamide or agarose can be suitably used as the main material constituting the electrophoresis gel.

  The electrophoresis gel can be formed by filling the gel holding region 4 of the gel cassette 1 with a gel solution containing a gel material and allowing it to stand to polymerize the gel solution, but the forming method is limited to this. Not. An electrophoretic gel prepared in advance may be accommodated in the gel holding region 4, and the electrophoretic gel and the resin layer 3 of the gel accommodating portion 2 may be brought into close contact with each other.

  By using the electrophoresis chip according to the present invention, it is possible to suitably perform from electrophoresis to sampling after electrophoresis, and by using a gel cassette 1 having a gel housing portion 2 made of synthetic resin, electrophoresis is possible. It is possible to prevent damage at the time of attaching to and detaching from the apparatus. In addition, the electrophoresis gel can be held in a state suitable for electrophoresis, and can be easily stored and distributed.

  The present invention is not limited to the above-described embodiments, and various modifications can be made within the scope shown in the claims. That is, embodiments obtained by combining technical means appropriately modified within the scope of the claims are also included in the technical scope of the present invention.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, this invention is not limited at all by these Examples.

[1. Materials and methods〕
(1-1. Hydrophilization treatment of gel cassette)
A gel cassette was prepared by an injection molding method using a cyclic olefin resin ZEONOR 1020 (manufactured by ZEON Corporation). The gel cassette was composed of two parts, a base material part having a depression that becomes a gel forming area and a lid part, and the depth of the gel forming area was about 1 mm. Next, the gel formation region was hydrophilized by oxygen plasma treatment or silicon oxide film coating.

The oxygen plasma treatment was performed as follows. The base part and lid part of the gel cassette were placed in a polymerization chamber of a plasma polymerization apparatus (manufactured by ULVAC), and the pressure was reduced until the ultimate vacuum in the polymerization chamber reached 1.5 × 10 ⁻³ Pascal. Next, oxygen was introduced into the polymerization chamber at a flow rate of 100 sccm, and plasma was generated at a plasma output of 500 W for 60 seconds to form a film. The contact angle of the gel-forming region was about 95 degrees before the treatment and about 10 degrees or less after the treatment.

Silicon oxide film coating was performed as follows. The base part of the gel cassette was placed in a polymerization chamber of a plasma polymerization apparatus (manufactured by ULVAC), and the pressure was reduced until the ultimate vacuum in the polymerization chamber reached 1.5 × 10 ⁻³ Pascal. Next, hexamethyldisiloxane (HMDS) and oxygen were simultaneously introduced into the polymerization chamber. The HMDS and oxygen flow rates were 5 sccm and 100 sccm, respectively, and plasma was generated at a plasma output of 300 W for 120 seconds to form a film. The contact angle of the gel-forming region was about 95 degrees before the treatment and about 10 degrees or less after the treatment.

(1-2. PVA coating)
The hydrophilized gel-forming region was coated using polyvinyl alcohol (PVA) of resins 1 to 4 shown below. Resin 1 (Wako Pure Chemical Industries, complete saponification type, average polymerization degree 500); Resin 2 (Wako Pure Chemical Industries, complete saponification type, average polymerization degree 2000); Resin 3 (SIGMA-ALDRICH, complete saponification type) Resin 4 (manufactured by Wako Pure Chemical Industries, partial saponification type, average polymerization degree 2000). These PVA were heated and dissolved in water to obtain an aqueous solution. Further, as a comparative example, an aqueous solution of carboxymethylcellulose sodium salt (Wako Pure Chemical Industries) and hydroxypropyl cellulose (Wako Pure Chemical Industries, 2.0 to 2.9 mPa / s) were used at a concentration of 5% by weight, respectively.

  Next, said resin was dripped at the concave gel formation area | region of a base material part, respectively, resin was apply | coated to the gel formation area | region surface using a spin coater (made by Mikasa Co., Ltd.), and it dried at 80 degreeC. Thereafter, a lid portion was attached to the substrate portion and ultrasonically welded to obtain an electrophoresis gel cassette.

(1-3. Preparation of electrophoresis gel)
After sealing the gel formation lower end part of the electrophoresis gel cassette produced in 1-2 above, the polyacrylamide gel solution was filled from the upper end part and allowed to stand to polymerize the gel. A polyacrylamide gel solution containing 13% acrylamide, 375 mM Tris-HCl / pH 8.8, 0.1% ammonium persulfate, and 0.1% tetraethylenediamine was used. 29.2% acrylamide-0.8% methylenebisacrylamide mixed solution was added to prepare a final concentration of acrylamide in the polyacrylamide gel of 13%.

(1-4. Sample)
As a sample to be subjected to SDS-PAGE, SeeBlue Pre-Stained Standard (manufactured by Invitrogen), which is a colored molecular weight marker, was used.

(1-5. Electrophoresis)
A mixture of the sample and an equal amount of 1% agarose was placed on the gel well of the electrophoresis gel cassette prepared in 1-3 above and subjected to SDS-PAGE. As the electrophoresis buffer, a cathode buffer containing 25 m Tris, 192 mM glycine and 0.1% SDS and an anode buffer containing 150 mM Tris-HCl / pH 8.8 were used. The electrophoresis gel cassette on which the sample was placed was solidified at 4 ° C. for about 5 minutes, mounted on the electrophoresis apparatus, 5 ml of the buffer was added to the buffer tank of the apparatus, and electrophoresis was performed at a constant current of 20 mA for 30 minutes.

(1-6. Observation of gel state after electrophoresis)
After the electrophoresis was completed, the electrophoresis gel cassette was removed from the electrophoresis apparatus, the lid was removed, and the gel was exposed to the atmosphere. The cassette from which the gel was exposed was placed in a constant temperature and humidity chamber at 20 ° C. and 75% RH, and the peeled state of the gel was visually observed.

[2. result〕
(2-1. Effect of hydrophilic treatment and PVA coating)
In order to examine the effect of hydrophilization treatment and PVA coating on the electrophoresis gel cassette, cassette 1 (cassette treated with silicon oxide film coating and PVA (resin 2) coating); cassette 2 (cassette treated only with silicon oxide film coating) ); Cassette 3 (cassette coated with silicon oxide film and PVA (resin 4)); electrophoresis was performed using cassette 4 (untreated cassette), and the peeled state of the gel after electrophoresis was observed. The results are shown in FIGS. FIG. 2 is a diagram showing a change in the peeled state of the gel with time in the cassette 1, and FIG. 3 is a diagram showing the gel state on the cassette 4. 4 is a diagram showing a change in the peeled state of the gel over time in the cassette 2, and FIG. 5 is a diagram showing a change in the peeled state of the gel in the cassette 3 over time.

  As shown in FIG. 2, in the cassette 1, the gel is satisfactorily held on the entire surface of the cassette until 30 minutes after the lid is removed and the gel is exposed, and from the side edge of the gel from 30 minutes to 150 minutes after the exposure. Although the part floated up a little, it was held on almost the entire surface of the cassette. Moreover, the electrophoresis pattern of the sample was also obtained well. On the other hand, as shown in FIG. 3, in the cassette 4, a good electrophoresis pattern was not obtained, and the gel was peeled off from the cassette shortly after the exposure. As shown in FIGS. 4 and 5, in cassettes 2 and 3, a good electrophoresis pattern was obtained, and the gel was well held on the entire surface of the cassette until 30 minutes after exposure, but 60 minutes after exposure. The side edge of the gel began to float and the gel peeled off 90 minutes after exposure.

  In addition, in the cassette subjected only to the oxygen plasma treatment instead of the silicon oxide film coating as a hydrophilic treatment, a good electrophoresis pattern was not obtained, and the gel peeled off from the cassette shortly after the exposure. Further, in the cassette subjected to the oxygen plasma treatment and the PVA (resin 2 or 4) coating treatment, a remarkable peeling prevention effect was not obtained.

  As described above, it was possible to prevent the gel from peeling by the PVA coating. Moreover, the cassette which performed the silicon oxide film coating as a hydrophilization treatment before PVA coating, and was coated using the completely saponified type PVA was optimal.

(2-2. Degree of polymerization of PVA)
In order to investigate the optimum degree of polymerization of PVA, electrophoresis gel cassettes were coated with solutions of PVA having different degrees of polymerization. The cassette was previously coated with a silicon oxide film, coated with a PVA solution containing 5% by weight of PVA of resins 1 to 3, and then subjected to electrophoresis, and the peeled state of the gel after electrophoresis was observed. The results are shown in FIG. 2, FIG. 6 and FIG. FIG. 6 is a diagram showing a change in the peeled state of the gel over time in a cassette coated with PVA having a polymerization degree of 500 (resin 1), and FIG. 7 is a coating with PVA having a polymerization degree of 4000 (resin 3). It is a figure which shows the change of the peeling state of the gel with progress of time in the done cassette. The change in the peeled state of the gel over time in the cassette coated with PVA (resin 2) having a polymerization degree of 2000 is the same as that shown in FIG.

  As shown in FIG. 6, when PVA having a polymerization degree of 500 is used, a good electrophoresis pattern is obtained, and the gel is well held on the entire surface of the cassette until 30 minutes after exposure. The side edges of the glass began to float and the gel peeled off 90 minutes after exposure. On the other hand, as shown in FIG. 2, when PVA having a polymerization degree of 2000 was used, a good electrophoresis pattern was obtained, and the gel was well held on the entire surface of the cassette until 60 minutes after exposure, and from 60 minutes to 150 minutes after exposure. Until then, the side edge of the gel was slightly lifted, but was held on almost the entire surface of the cassette. Further, as shown in FIG. 7, when PVA having a polymerization degree of 4000 was used, a good electrophoresis pattern was obtained, and the gel was well held on the entire surface of the cassette until 60 minutes after exposure, and from 60 minutes to 180 minutes after exposure. Until then, the side edge of the gel was slightly lifted, but was held on almost the entire surface of the cassette.

(2-3. PVA coating range)
In order to examine the optimal coating range of PVA, electrophoresis gel cassettes with different coating ranges with PVA were prepared. Using a cassette previously coated with a silicon oxide film, the non-coated portion was masked with Kapton tape, and then PVA (resin 2) was spin-coated and dried, and partially coated with PVA. Electrophoresis was performed from the outermost periphery of the gel formation region using a cassette coated with PVA coating on the periphery of 3 mm width, 5 mm width, or 7 mm width, and the peeled state of the gel after electrophoresis was observed. The results are shown in FIGS. FIG. 8 is a graph showing changes in the peeled state of the gel over time in a cassette coated with PVA with a width of 3 mm, and FIG. 9 is a graph showing the change of the gel over time in a cassette coated with PVA with a width of 5 mm. FIG. 10 is a diagram showing changes in the peeled state of the gel over time in a cassette with a PVA-coated cassette having a width of 7 mm around.

  As shown in FIG. 8, in the cassette coated with PVA having a width of 3 mm, a good electrophoretic pattern was obtained and the gel was well retained on the entire surface of the cassette until 30 minutes after the exposure. The side edges of the gel began to float and the gel peeled off significantly 120 minutes after exposure. As shown in FIGS. 9 and 10, in a cassette coated with PVA at a width of 5 mm or 7 mm, a good electrophoretic pattern was obtained, and the gel was well held on the entire surface of the cassette until 60 minutes after exposure. From 60 minutes to 180 minutes, the side edge of the gel slightly floated, but was held on almost the entire surface of the cassette.

  As described above, even in the electrophoresis gel cassette in which the gel formation region is partially PVA-coated, the same peeling prevention effect as that in the electrophoresis gel cassette in which the gel formation region is entirely PVA-coated is obtained.

(2-4. Coating with other resins)
In order to examine the effect of coating with a resin other than PVA, an electrophoresis gel cassette was coated with carboxymethylcellulose (CMC) and hydroxypropylcellulose (HPC). The cassette was previously coated with a silicon oxide film, coated with a solution containing 5% by weight of each resin and then subjected to electrophoresis, and the peeled state of the gel after electrophoresis was observed.

  As a result, in the cassette coated with CMC or HPC, the separation pattern became unclear compared with the PVA coating cassette, and the effect of preventing the gel from peeling was not sufficiently obtained.

  As described above, by coating a part of the surface of the electrophoresis gel cassette in contact with the water-soluble resin, particularly when the water-soluble resin is a completely saponified PVA, the PVA swells in water. It is considered that viscosity can be exhibited and gel peeling can be prevented.

  By using the present invention, it is possible to keep the state of the gel after electrophoresis favorable and to perform sampling from the gel more easily, so that research on biopolymers such as DNA, RNA, and protein is further developed. In particular, it can contribute to the development of industries in the fields of medicine, biology and chemistry.

FIG. 1 is a schematic view showing an electrophoresis support holder according to an embodiment of the present invention. FIG. 2 is a diagram showing the state of the gel after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention. FIG. 3 is a diagram showing a comparative example of the present invention. FIG. 4 is a diagram showing a comparative example of the present invention. FIG. 5 is a diagram showing the state of the gel after electrophoresis using the electrophoresis support holder according to one embodiment of the present invention. FIG. 6 is a diagram illustrating a state of the gel after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention. FIG. 7 is a diagram showing a gel state after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention. FIG. 8 is a diagram illustrating a state of the gel after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention. FIG. 9 is a diagram illustrating a state of the gel after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention. FIG. 10 is a diagram illustrating a state of the gel after electrophoresis using the electrophoresis support holder according to the embodiment of the present invention.

Explanation of symbols

1 Gel cassette (Electrophoresis support holder)
2 Gel container (container)
2a Base part 2b Lid part 3 Resin layer 4 Gel holding area

Claims (8)

An electrophoretic support holder having an accommodating portion for accommodating an electrophoretic support therein and having a detachable lid,
Have a resin layer containing at least a portion in a water-soluble resin surface in contact with the support for electrophoresis of the accommodating portion,
The electrophoretic support holder , wherein the water-soluble resin is a completely saponified resin .   The water-soluble resin is a water-soluble polymer derived from synthesis such as polyvinyl alcohol, polyvinyl alcohol (PVA) polyacrylic acid, polymethacrylic acid, polyvinylamine, polyallylamine, starch, cellulose dielectric, pectin, alginic acid, agarose, pullulan. The support member for electrophoresis according to claim 1, which is selected from the group consisting of a polysaccharide such as gelatin and a protein such as gelatin.   The resin layer is formed on an outer peripheral portion of a bottom portion of the housing portion, and has an area of at least 5 to 100% of an area of a surface of the housing portion that contacts the support for electrophoresis. The support holder for electrophoresis according to claim 1 or 2.   The said resin layer is formed in the whole inner surface of the bottom part of the said accommodating part, The support body holder for electrophoresis of any one of Claims 1-3 characterized by the above-mentioned.   The support member for electrophoresis according to any one of claims 1 to 4, wherein the container is made of a synthetic resin.   The support member for electrophoresis according to any one of claims 1 to 5, wherein a surface of the housing portion that contacts the support for electrophoresis is hydrophilized. The degree of polymerization of the water-soluble resin is 2000 to 4000, The support holder for electrophoresis according to any one of claims 1 to 6. Electrophoresis chip comprising a electrophoresis support holder according, the electrophoresis support housed inside a support for the holder the electrophoresis to any one of claims 1-7.