JPS61296256A - Electrophoretic medium material and its preparation - Google Patents

Abstract

PURPOSE:To attain to enhance the adhesiveness between a support and a medium layer, by providing a three-layered structure consisting of a plastic support layer, an adhesive layer and an electrophoretic medium layer (polyacrylamide gel film) containing at least one carbamoyl group. CONSTITUTION:A plastic support film and an electrophoretic medium layer (polyacrylamide gel film) are adhered by an adhesive layer formed of an amido group-containing grafted derivative of a methyl methacrylate macromonomer to form a three-layered structure. If this three-layered structure is formed, said three-layered structure becomes hard to separate even by various operations in the process for drying the electrophoretic medium layer after electrophoresis.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of the Invention] The present invention relates to an electrophoretic medium material and a method for producing the same. It relates to media materials and their manufacturing methods.

[Background of the Invention] In DNA and RNA base sequencing methods based on the Bostravel method, slab electrophoresis using a polyacrylamide gel membrane is an essential operation. In particular, as gene-related research progresses in recent years, there is an urgent need to speed up DNA base sequencing operations.

Polyacrylamide gel membranes are obtained by crosslinking and polymerizing monomers such as acrylamide using a difunctional crosslinking agent such as N,N'-methylenebisacrylamide in the presence of a catalyst.

Note that when forming this polyacrylamide gel film, a denaturing agent such as urea or formamide is usually included.

The above polymerization reaction is a radical crosslinking polymerization, and since the reaction is inhibited by the influence of oxygen, the polyacrylamide gel film must be prepared in a state where oxygen is blocked. For this reason, polyacrylamide gel membranes are now generally produced by injecting a gel-forming solution into a cell (having a certain space, e.g. about 0.3 to 1 mm) formed by two glass plates, and then introducing oxygen into the cell. A gel film is formed by cross-linking polymerization in a blocked state.

Since this method forms a gel film between two glass plates, it has major drawbacks such as poor handling and difficulty in mass-producing the gel film.

Electrophoresis using the polyacrylamide gel membrane formed as described above is carried out, for example, as follows.

The polyacrylamide gel membrane is held vertically between glass plates, and after pre-electrophoresis, a sample (for example, Maxam-Gilbert digested 32 p-labeled DNA) is injected at regular intervals, and then electrophoresis is performed. After performing electrophoresis for a certain period of time (e.g., about 6 to 12 hours), one side of the glass plate is carefully removed, and the exposed surface is covered with a synthetic resin film such as polyvinylidene chloride film. This is used to perform autoradiography processing.

That is, an X-ray film and an intensifying screen are sequentially placed on the film covering the polyacrylamide gel membrane, and exposure is performed at a low temperature (for example, -80°C) for a certain period of time (for example, about 10 to 20 hours). After the exposure, the base sequence of the DNA can be determined by autoradiography processing, which consists of developing the X-ray film and reading the separation migration pattern of the DNA.

Since autoradiography processing requires a long time as described above, it is desired to speed up the processing, and furthermore, it is desired to improve the resolution of reading.

Conventional electrophoresis media materials using acrylamide gel membranes use glass plates as described above, which poses problems in handling, and experiments often fail due to the glass breaking. Furthermore, during autoradiography,
As mentioned above, it is necessary to remove the glass on one side, and the gel film may be damaged at this time, so this operation requires great skill. Therefore, it is eagerly awaited to develop a polyacrylamide gel membrane that has improved handling properties and has improved handling properties.

[Summary of the Invention] An object of the present invention is to use a plastic film (or sheet) as a support instead of a glass plate, and to improve the adhesiveness between the support and the electrophoresis medium layer (polyacrylamide gel membrane). An object of the present invention is to provide an electrophoretic medium material and a method for producing the same.

The present invention consists of an electrophoretic medium material having a three-layer structure consisting of the following layers in sequence: [I] Plastic support layer; [11] Methyl methacrylate macromonomer containing an amide group an adhesive layer formed from a grafted derivative; and.

[m] An electrophoretic medium layer that is an aqueous polyacrylamide gel obtained by crosslinking polymerizing an acrylamide compound and a crosslinking agent in the presence of water, and containing a compound having at least one carbamoyl group as a modifier.

In the electrophoresis medium material described in L, an intermediate layer containing an amide group-containing grafted derivative of a methyl methacrylate macromonomer is provided on a plastic support, and then an acrylamide-based compound and a crosslinking agent are formed on the intermediate layer. It can be produced by a method in which a polyacrylamide-based aqueous gel electrophoresis medium layer is formed by cross-linking polymerization in the presence of water and a compound having at least one carbamoyl group (□) as a modifier.

As described above, the electrophoretic medium material of the present invention connects the support layer and the electrophoretic medium layer (polyacrylamide gel membrane) with an adhesive layer formed from a grafted derivative containing a 7-amide group of methyl methacrylate macromonomer. The three-layer structure is difficult to separate even by various operations in the drying process of the electrophoresis medium layer after electrophoresis, so the electrophoresis medium layer is The advantage is that very little damage occurs and therefore there is no need to use auxiliary supports such as filter paper.

Furthermore, the electrophoretic medium material of the present invention is prepared by forming a layer containing an amide group-containing grafted derivative of methyl methacrylate macromonomer on the support in a horizontal apparatus, and then forming an electrophoretic medium layer thereon. Since it can also be manufactured by the method described above, it greatly contributes to the mass production of electrophoretic media materials.

[Detailed Description of the Invention] The support for the electrophoresis medium material of the present invention is preferably a plastic sheet (or film).
This plastic sheet may be made of any plastic. Examples of preferred plastic sheets include hydrophilic polymers or polymers whose surfaces have been made hydrophilic by known surface treatments (e.g.
Polyethylene terephthalate, polycarbonate of bisphenol A, polyvinyl chloride, vinylidene chloride/vinyl chloride copolymer, polymethyl methacrylate, polyethylene, polypropylene, cellulose acetate,
cellulose acetate propionate, etc.) films,
Examples include molded products such as plates or sheets. In particular, it is preferable to use a sheet made of polyethylene terephthalate.

For the treatment to make the surface of these polymer moldings hydrophilic, known methods such as ultraviolet irradiation, glow discharge treatment, corona discharge treatment, flame treatment, electron beam irradiation, chemical etching, electrolytic etching, etc. are applied. be able to.

The support generally has a thickness of about 50 pm to about 5007 tm,
Preferably, the size of about 70 m to about 300 m is used.

In the present invention, an adhesive layer is formed on seven of the supports.

The adhesive layer is a layer formed from an amide group-containing grafted derivative of methyl methacrylate macromonomer. A macromonomer is generally defined as a polymerizable monomer with a relatively high molecular weight substituent having a certain repeating unit, and a methyl methacrylate macromonomer derivative is a polymerizable methyl methacrylate group derivative. It is a macromonomer containing in the molecule. The amide group-containing grafted derivative of methyl methacrylate macromonomer is obtained by grafting methyl methacrylate macromonomer onto a monomer containing an amide group and one ethylenically unsaturated double bond, preferably an amide group-containing acrylic acid or methacrylic acid compound. It is a compound obtained by treating the methyl methacrylate macromonomer with one or more types to initiate graft copolymerization with the monomer at the methyl methacrylate group end of the methyl methacrylate macromonomer. Examples of amide group-containing acrylic acid compound monomers include acrylamide, N
-Amide group-containing methacrylic acid compounds such as methylacrylamide, N,N-dimethylacrylamide, diacetone acrylamide, N-methylolacrylamide, etc./
Examples of the mer include methacrylamide, N-methylmethacrylamide, and the like.

Synthesis examples of typical amide group-containing grafted derivatives of methyl methacrylate macromonomers used in the present invention are described below. In addition, in each synthesis example, 1 part' and 1 part
%] means r parts by weight j and "% by weight", respectively.

[Synthesis Example 1] A mixed solvent of 17.5 parts of acetone and 82.5 parts of toluene was charged into a glass flask equipped with a stirrer, a reflux condenser, a dropping funnel, a thermometer, and an N2 gas inlet, and N2 was introduced. Methyl methacrylate (hereinafter MMA) was added under reflux while
), 3.2 parts of thioglycolic acid (hereinafter abbreviated as TGA) as a chain transfer agent, and 3 parts of azobisisobutyronitrile as a polymerization initiator were continuously added dropwise over 3 hours. Polymerization was performed. Further after that 2
Polymerization was completed by heating for a period of time to obtain a solution of the polymer having the following structural formula [I]. 6 N-hexane was added to a portion of the reaction solution to form a precipitate, and this precipitate was dried under reduced pressure. The acid value was measured and found to be 0.340 mg equivalent/g.

C00CH3 After distilling off a portion of acetone from the above reaction solution, 0.5% of triethylamine (catalyst), 200 ppm of hydroquinone monomethyl ether (polymerization inhibitor), and 1.3 times the molar amount of glycidyl methacrylate relative to the acid were added thereto. 1
The reaction was carried out at a reaction temperature of 110° C. for 4 hours. The reaction rate determined from the decrease in acid value was 96%.

The above reaction solution was poured into 10 volumes of n-hexane to form a precipitate, and this precipitate was dried under reduced pressure at 80°C to obtain 95 parts of a macromonomer of the following structural formula [II]. The polystyrene equivalent molecular weight of this Makomo/Marr by gel permeation chromatography (GPC) is 2B4.
0 (number average) and 6.470 (weight average). Moreover, the hydroxyl value was 0.350 mg equivalent/g.

[Synthesis Examples 2 to 4] Maccomo obtained in Synthesis Example 1 was placed in the same apparatus as Synthesis Example 1.
30 parts of diacetone acrylamide (hereinafter abbreviated as MMA), 50 parts of diacetone acrylamide (hereinafter abbreviated as DAA), diacetone acrylamide (hereinafter abbreviated as IAA)
20 parts of methyl ethyl ketone and 3.0 parts of azobisisobutyronitrile were charged, and the mixture was reacted for 800 hours under reflux (approximately 80°C) with N2 gas introduced.

This reaction solution was poured into 10 times the amount of n-hexane, precipitated, and dried at 80° C. under reduced pressure to obtain 94 parts of comb-shaped graft copolymer (P-1). The obtained graft copolymer showed a single peak by GPC and had a number average molecular weight of about 19,000.

By the same operation, the composition ratio of the component monomers of the graft copolymer was changed, and two types of comb-shaped graft copolymers (P-
Table 1 shows the respective comb-shaped graft copolymers synthesized.

Table 1 Synthesis Example (A) Copolymerization Number average Yield: Molecular weight: 2 P-I MMAM 30 parts Approximately 1!3
．． 000 92 part MMA 50ffl! DAA 20 copies 3 P-2MMAM 30 copies Approx. 22,000
92 copies MMA 30 copies DAA 40 copies 4 P-3MMAM 30 copies Approx. 12.000
95 parts M) IA 30 parts (B) 40 parts Note) (A) Niger raft copolymer MMAM: Methyl methacrylate macromonomer (Synthesis example 1) MMA + methyl methacrylate DAA Nidiacetone acrylamide (B):
N,N-dimethylacrylamide These amide group-containing grafted derivatives of methyl methacrylate macromonomers may be used alone or in combination with other polymers as long as they account for up to 50% by weight of the layer-forming components. May contain substances and additives. Examples of such polymeric substances and additives include polymers such as diacetyl cell, nitro cell, polyvinyl alcohol, polyacrylamide, polymethyl methacrylate, and polyvinylidene chloride, and polyol compounds such as glycerin.

The adhesive layer formed from the amide group-containing grafted derivative of methyl methacrylate macromonomer can be provided on the surface of the support using a known coating method.

That is, when the macromonomer conductor is water-soluble or hydrophilic, an aqueous solution thereof or a water/organic solvent mixed solvent solution containing water as a main component is applied onto a support by a known method and dried. A method of forming an adhesive layer can be used. When the macromonomer derivative is hydrophobic and water-insoluble, a solution of the macromonomer derivative in an organic solvent or a mixed solvent of an organic solvent and water containing an organic solvent as a main component is coated on a support by a known method, A method of drying to form an adhesive layer can be used.

Examples of organic solvents that can be used include acetone,
Ketones such as methyl ethyl ketone; Alcohols such as methanol and ethanol: N.

Examples include ethers such as N-dimethylformamide; dimethylsulfonate dimethyl ether and dioxane.

The adhesive layer formed from the amide group-containing grafted derivative of the methyl methacrylate macromonomer is desirably formed from the amide group-containing grafted derivative of the methyl methacrylate macromonomer alone or from a composition containing 80ii% or more of the derivative. It is desirable that the layer be formed only from an amide group-containing grafted derivative of methyl methacrylate macromonomer.

The thickness of the adhesive layer after drying is about 0.17 parts m to about 3 parts m.
, preferably in the range of about 0.2 pm to about 2 pm.

Next, the electrophoresis medium layer (hereinafter also referred to as gel medium layer, polyacrylamide gel membrane, or simply gel membrane)
I will explain about it.

Polyacrylamide gel membrane is an aqueous gel obtained by dissolving or dispersing an acrylamide compound and a crosslinking agent in water as an aqueous solution or dispersion to prepare a gel forming liquid, and then crosslinking and polymerizing both in the liquid. 6 In this specification, unless otherwise specified, dissolution (in water) and dissolution (in water) are used.
The term ``dissolution (in water)'' includes both dispersion, and the term ``aqueous solution'' includes both an aqueous solution and an aqueous dispersion, and also includes a mixture of an organic solvent and water that may be added as a solvent or a dispersion medium if desired. do.

Acrylamide compounds that can be used to form polyacrylamide membranes include acrylamide 2.
N-methylacrylamide, N,N-dimethylacrylamide, N-(hydroxymethyl)acrylamide,
Examples include acrylamide-based compounds such as diacetone acrylamide) and methacrylamide-based compounds such as methacrylamide, and these compounds can be used alone or in combination of two or more. Among these acrylamide compounds, acrylamide is the most preferred;
It is also preferable to use one or more of acrylamide and other acrylamide-based compounds or methacrylamide-based compounds in combination.

As a crosslinking agent, “Electrophoresis (Electr
ophoresis) J 1981.2, 213-
228 etc. (one type or a combination of two or more types) can be used. A specific example of the crosslinking agent is N,N'-methylenebisacrylamide (BIS).
, N,N'-propylene bisacrylamide (PBA)
, di(acrylamide dimethyl) ether (DAE),
1.2-diacrylamide ethylene glycol (DEG
); ethylene ureabisacrylamide (EUB), ethylene diacrylate (EDA), N,N'-diallyltardiamide (N,N'-diallyltar
tardiamide: DATD); and N, N'-
Bisacrylylcystamine (N,N'-bigacry
Bifunctional compounds such as lylcystamine, BAC) and the like can be mentioned.

The amount of crosslinking agent is approximately 0.0% based on the total weight of monomer and crosslinking agent.
1 wt% to about 30 wt%, preferably about 0.5 wt%
% to about 10 wt%.

As for the gel concentration, Nis Hijaten (S.

Arch, Biochem, Biap
hys-) J 1 (Addendum).

147 (1862), relative to the volume of the gel film consisting of monomer, crosslinker and water,
The amount of monomer and crosslinker is about 3 w/v% to about 30
It is preferably used within the w/v% range.

Examples of the modifier to be included in the polyacrylamide gel membrane include compounds having at least one carbamoyl group, specific examples of which include urea, formamide, and the like. Among these, urea is particularly preferably used. The amount of modifier is from about 40 wt/v% to about 6%, based on the volume of the aqueous gel containing the monomer and crosslinker.
It is used in the range of 0 wt/v%. When using urea, the amount is from about 6 mol (about 360 g) to a saturated dissolution amount, preferably from about 7 mol (about 420 g) to a saturated dissolution amount per 1 aqueous gel containing a monomer and a crosslinking agent. It can be used within the range of.

The polyacrylamide gel membrane can contain a water-soluble polymer.

As the water-soluble polymer, addition polymerization type or condensation polymerization type water-soluble polymer can be used.

Specific examples of addition polymers include nonionic water-soluble polymers such as polyvinyl alcohol, polyvinylpyrrolidone, and polyacrylamide. Specific examples of polycondensation polymers include nonionic water-soluble polyalkylene glycols such as polyethylene glycol and polypropylene glycol. The molecular weight of the water-soluble polymer is preferably in the range of about 10,000 to about 100,000. Among these water-soluble polymers, polyethylene glycol and polyacrylamide are particularly preferred.

The water-soluble polymer ranges from about 2 wt% to about 100 wt%, preferably about 5 wt% based on the total weight of monomer and crosslinker.
It is used in a range of wt% to about 50wt%. By adding a water-soluble polymer, the polyacrylamide gel film has plasticity, so it will not break during cutting, and the gel film will also have plasticity when drying, improving its brittleness and making it less likely to break. There is an advantage in that. Furthermore, the viscosity of the gel film can be controlled by selecting the molecular weight and amount of the water-soluble polymer added.

The polyacrylamide gel membrane can contain agarose. The agarose is not particularly limited as long as it is a known agarose, and low electroosmotic, medium electroosmotic, and high electroosmotic agaroses can also be used. An example of agarose that can be used is JP-A-55-
No. 5730, JP-A-55-110946, JP-A-57
Examples include agarose disclosed in publications such as No.-502098.

The amount of agarose added is about 0.2 w/v% to about 2 w/v% with respect to the end of the gel composition containing the monomer and the crosslinking agent.
v%, preferably about 0.3 w/v% to about 1.2 w/v%
It is used at a ratio of v%. By adding agarose to the gel film, it is possible to control the viscosity of the solution to an appropriate level by changing the temperature of the gel forming solution, and its fluidity can be stopped. This has the advantage of being easier to do.

The polyacrylamide gel membrane can contain a pH 11 buffer. As a buffer, pH 8.0 to i
Any buffering agent capable of buffering to a pH value within the range of pH 8.0 to 9.0 can be used. As buffering agents that can be used,
Published in 1986') Pages 1312-1320: 'Data
Fore Biochemical ° Research (Data for
Biochemical Research) J
(Edited by R-ni C, Dawson at a1 2nd edition, Oxford at the Clarendon
Press, published in 19813) Pages 47B-508: rBiochemistry (Biochemistr7
) J, 5.487 (1968); and rAnalyt Biochemistry (Analyt.
ical Biochemistry) j 104
．． Examples include buffers described in publications such as 300-310-(1980).

A specific example thereof is tris(hydroxymethyl)aminomethane (Tris) [GASR
Registry No.7? -86-11, N, N-
Bis(2-hydroxyethyl)glycine
), N-2-hydroxyethylpiperazine-No
Na of -2-hydroxypropane-3-sulfonic acid
Salts, etc., Na salts of N-2-hydroxyethylpiperazine-No-3-propanesulfonic acid, salts of N-[tris(hydroxymethyl)methyl]-3-aminopropanesulfonic acid, etc. Examples include Na salts or Na salts, and acids, alkalis, or salts combined with any of these if necessary. Examples of particularly preferred buffers include Tris, boric acid and ED.
There is a combination of TA-2Na salts (pH 8,3).

In the present invention, the polyacrylamide gel membrane is a monomer typified by acrylamide, a difunctional allyl (ally)
l) It is obtained by radical crosslinking polymerization of a monomer and a crosslinking agent in an aqueous solution in which a compound or an acrylic compound (crosslinking agent), a water-soluble polymer, and agarose are substantially uniformly dissolved. It is estimated that the water-soluble polymer and agarose are substantially dispersed in the three-dimensional cross-linked polymer formed from the body and the cross-linking agent, and the polymer chains of the latter two are entangled with the three-dimensional cross-linked polymer. This structure is a characteristic of the gel medium.

The above radical crosslinking polymerization reaction can be caused in the absence of molecular oxygen in the presence of peroxide and/or by known methods such as ultraviolet irradiation. This reaction can also be accelerated by heating and UV radiation.

As a catalyst for radical crosslinking polymerization, "Electrophoresis" 198
1.2°213-219. 1981.2.220-2
28; Aomizu, Nagai eds. “Latest Electrophoresis Methods J” (197
It is possible to appropriately select and use the known low-temperature radical polymerization initiators described in the following publication (published in 2010). Specific examples of preferred radical polymerization initiators include β-dimethylaminopropionitrile (DMAPN)-ammonium peroxonisulfate mixture, N,N,N',N'-tetramethylethylenediamine (TEMED)-ammonium peroxonisulfate mixture, and TEMED-riboflavin. Examples include combinations of mixtures, TEMED-riboflavin-hydrogen peroxide mixtures, and ultraviolet irradiation. The content of the radical polymerization initiator is approximately 0.0% based on the total weight of the monomer and crosslinking agent.
It ranges from 3% to about 5% by weight, and preferably from about 0.5% to about 3% by weight.

The gel medium layer is provided by coating a gel forming liquid by a known method on the layer containing the above-mentioned methacrylate macromonomer derivative provided on a support having a smooth surface, and then crosslinking and polymerizing the gel forming liquid. By doing so, it can be formed into layers.

When cross-linking and polymerizing the gel-forming liquid on the surface of the support,
The gel-forming liquid can be further covered with a covering material such as a cover film, sheet or plate. The cover film, sheet, or plate used for this purpose may be made of the same material as the support. The thickness of this coating material is less than or equal to 300 ILm;
The practically preferred range is about 84 m to about 200 pm.
, a particularly preferable range is about 10 gm to about 100 hirom
It is.

The polyacrylamide gel film may contain other additives such as an antioxidant, if necessary. As the antioxidant, various compounds known to be incorporated into the gel film can be used. Specific examples of the antioxidant include dithiothreitol and 2-mercaptoethanol.

Other additives include wetting agents, and the polyacrylamide gel film can also contain polyol compounds such as glycerin and ethylene glycol. The content of the polyol compound is approximately 5 w/w/6 for the gel membrane.
It is selected from the range from v% to approximately 40 w/v%. Among polyol compounds, glycerin is particularly preferred. By adding a wetting agent, it is possible to prevent the gel film from drying out due to extreme water evaporation during storage, and it also improves the physical properties of the gel film, such as preventing brittleness caused by extreme dryness and preventing cracking. It has the advantage of

The gel medium material for electrophoresis of the present invention can be used in both horizontal and vertical plate electrophoresis, disk electrophoresis, etc. according to the known methods described in the above-mentioned documents.

Next, examples of the present invention will be shown.

[Example 1] A polyethylene terephthalate (PET) sheet (support) whose surface was made hydrophilic by ultraviolet irradiation treatment was coated with about 0.
A coating liquid (Table 2) obtained by dissolving the amide group-containing grafted derivative of the methacrylate macromonomer obtained in the above synthesis example in a7ton was applied to a thickness (solid content) of 5 gm, It was dried at about 110° C. to form an amide group-containing grafted derivative layer (adhesive layer) of the macromonomer.

In addition, P-1 to F-3 in Table 1 are Synthesis Examples 2 to 4.
It was created in

Table 2 Composition sample of coating liquid for adhesive layer formation Coating liquid concentration of macromonomer Amide group content number Grafted derivative (g/100ml solvent)
I MMAM (comparison sample) 5gF-1
5g F-25g F-35g First, the adhesiveness between the PET sheet (support) and the adhesive layer was evaluated by a cross-cut method. As a result, samples 1~
The adhesive layer adhered uniformly and strongly to the support in all samples 4, and particularly in samples 2 to 4 (samples according to the present invention), the adhesive layer adhered particularly strongly to the support.

On each adhesive layer provided on the support, 87 g of acrylamide 1 1 , 630 mg of BIS, 42 g of urea, and tris(hydroxymethyl)aminomethane [CAS R
egiStry No. 77-88-1 1 1,
0 8g, boric acid 0, 55g, and EDTA-2'
Ammonium belloxonisulfate (5% by weight) was added as a polymerization initiator to a 100ml solution containing 93mg of Na salt.
, 3mjL. TEMED33gJl added to 0
．． It was molded to a thickness of 5 mm to form a polyacrylamide gel film.

In addition, PE was prepared in the same manner except that no separate adhesive layer was provided.
A comparative sample was prepared by forming a polyacrylamide gel film on a T-sheet.

The obtained gel film was pressed with a finger to evaluate the adhesion between the gel film and the support. As a result, sample 1 (comparative sample) sample 2~
In the case where No. 4 (sample according to the present invention) was used as the adhesive layer, the gel film adhered uniformly and firmly to the support (excellent adhesion), but the comparative sample had poor adhesion.

[Example 2] A PET sheet was prepared with the same adhesive layer as in Table 2 of Example 1, and acrylamide 11.8 was applied on this adhesive layer.
7g, BIS630mg, Agarose 1600 (manufactured by Wako Tsumugi Pharmaceutical ■) 0.3g, polyacrylamide 2.5g, tris(hydroxymethyl)aminomethane 1.08g, boric acid 0.55g and EDTAφ2Na salt 93mg
Ammonium belloxodisulfate (5% by weight) 1.3 mu as a polymerization initiator was added to a 100 mJ solution consisting of
The material to which MED33JLu was added was molded to a thickness of 0.5 mm to form a polyacrylamide gel film to obtain four types of samples (samples 5 to 8).

A comparative sample was also prepared by directly forming the above polyacrylamide gel film on a PET sheet.

Using this gel membrane, a sample obtained by Maxam-Gilbert digestion of P-DNA was subjected to electrophoresis and a DNA base sequencing experiment was performed.After electrophoresis, the gel membrane was subjected to deurea and DNA fixation. For this purpose, the gel film was immersed in a 10% acetic acid aqueous solution for 1 hour.The gel film was dried and then subjected to autoradiography according to a conventional method.

The state of adhesion between the support and the gel membrane was observed when the gel membrane was immersed in a 10% aqueous acetic acid solution.

The gel film of the comparative sample was completely peeled off from the support.

On the other hand, the gel films formed on the adhesive layers of samples 5 to 8 were
All were completely adhered to the support. After further shaking the gel film for 1 hour, we observed the state of adhesion between the gel film and the support, and found that in Sample 1, part of the edge of the gel film had peeled off from the support, but in Samples 6 to 8 ( In the sample of the present invention), the gel film remained uniformly adhered to the support. Separately, after drying the gel membrane, the state of adhesion between it and the support was observed, and it was found that the gel membranes of these samples all adhered well to the support. Furthermore, even after autoradiography treatment, the gel film remained uniformly adhered to the support.
There were no problems with adhesion and good results were shown.

Claims (1)

[Claims] 1. An electrophoretic medium material comprising a three-layer structure in which the following layers are sequentially laminated: [I] Plastic support layer; [II] Grafted derivative containing an amide group of methyl methacrylate macromonomer an adhesive layer formed from; and [III] a polyacrylamide-based aqueous gel formed by crosslinking polymerization of an acrylamide-based compound and a crosslinking agent in the presence of water;
An electrophoretic medium layer comprising a compound having at least one carbamoyl group as a modifier. 2. The electrophoresis medium material according to claim 1, wherein the medium layer further contains a water-soluble polymer and agarose. 3. The electrophoresis medium material according to claim 1 or 2, wherein the denaturing agent is urea or formamide. 4. The electrophoretic medium material according to claim 1 or 2, wherein the support layer is made of a plastic sheet. 5. The electrophoretic medium material according to claim 4, wherein the plastic sheet is made of polyethylene terephthalate. 6. A patent claim characterized in that the adhesive layer formed from the amide group-containing grafted derivative of methyl methacrylate macromonomer is a layer formed substantially only from the amide group-containing grafted derivative of methyl methacrylate macromonomer. The electrophoresis medium material according to item 1. 7. After providing an intermediate layer containing an amide group-containing grafted derivative of a methyl methacrylate macromonomer on a plastic support, at least one acrylamide-based compound and a crosslinking agent are used as modifiers on the intermediate layer. 1. A method for producing an electrophoresis medium material, which comprises cross-linking polymerizing a compound having a carbamoyl group in the presence of water to form a polyacrylamide-based aqueous gel electrophoresis medium layer. 8. The method for producing a medium material for electrophoresis according to claim 7, wherein the medium layer further contains a water-soluble polymer and agarose. 9. The method for producing a medium material for electrophoresis according to claim 7, wherein the denaturing agent is urea or formamide.