US20020079222A1 - Composition for use as an electrophoretic medium - Google Patents
Composition for use as an electrophoretic medium Download PDFInfo
- Publication number
- US20020079222A1 US20020079222A1 US10/078,401 US7840102A US2002079222A1 US 20020079222 A1 US20020079222 A1 US 20020079222A1 US 7840102 A US7840102 A US 7840102A US 2002079222 A1 US2002079222 A1 US 2002079222A1
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- Prior art keywords
- cassette
- gel
- electrophoretic
- electrophoretic medium
- electrophoresis
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Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44747—Composition of gel or of carrier mixture
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C51/00—Shaping by thermoforming, i.e. shaping sheets or sheet like preforms after heating, e.g. shaping sheets in matched moulds or by deep-drawing; Apparatus therefor
- B29C51/26—Component parts, details or accessories; Auxiliary operations
- B29C51/30—Moulds
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C2791/00—Shaping characteristics in general
- B29C2791/004—Shaping under special conditions
- B29C2791/006—Using vacuum
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C2793/00—Shaping techniques involving a cutting or machining operation
- B29C2793/0009—Cutting out
- B29C2793/0018—Cutting out for making a hole
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C35/00—Heating, cooling or curing, e.g. crosslinking or vulcanising; Apparatus therefor
- B29C35/16—Cooling
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C51/00—Shaping by thermoforming, i.e. shaping sheets or sheet like preforms after heating, e.g. shaping sheets in matched moulds or by deep-drawing; Apparatus therefor
- B29C51/08—Deep drawing or matched-mould forming, i.e. using mechanical means only
- B29C51/082—Deep drawing or matched-mould forming, i.e. using mechanical means only by shaping between complementary mould parts
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C51/00—Shaping by thermoforming, i.e. shaping sheets or sheet like preforms after heating, e.g. shaping sheets in matched moulds or by deep-drawing; Apparatus therefor
- B29C51/10—Forming by pressure difference, e.g. vacuum
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
- B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
- B29C51/00—Shaping by thermoforming, i.e. shaping sheets or sheet like preforms after heating, e.g. shaping sheets in matched moulds or by deep-drawing; Apparatus therefor
- B29C51/26—Component parts, details or accessories; Auxiliary operations
- B29C51/42—Heating or cooling
- B29C51/428—Heating or cooling of moulds or mould parts
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Pathology (AREA)
- Electrochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Mechanical Engineering (AREA)
- Dispersion Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Laminated Bodies (AREA)
- Adhesives Or Adhesive Processes (AREA)
- Sampling And Sample Adjustment (AREA)
- Thermotherapy And Cooling Therapy Devices (AREA)
- Addition Polymer Or Copolymer, Post-Treatments, Or Chemical Modifications (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
- Electrochromic Elements, Electrophoresis, Or Variable Reflection Or Absorption Elements (AREA)
Abstract
The present invention is concerned with a disposable electrophoresis cassette particularly suitable for pre-cast polyacrylamide gels for protein and nucleic acid electrophoresis. The invention also comprises a support plate for the said cassette that acts as a heat sink and provides a more uniform migration front in operation since the temperature of the gel is substantially the same during electrophoresis operation. Also disclosed is a novel comb element for filling the cassette, and a novel method therefor.
Description
- This application is a divisional of the application Ser. No. 09/387,758 filed on Sep. 1, 1999.
- (a) Field of the Invention
- The present invention is concerned with a disposable electrophoresis cassette particularly suitable for pre-cast electrophoretic media for protein and nucleic acid electrophoresis. The invention also comprises a support plate for the said cassette to ensure proper rigidity thereof into the electrophoresis apparatus, and acting as a heat sink, thus providing a more uniform migration front in operation because the temperature of the medium is substantially even anywhere inside the cassette during electrophoresis operation. The invention also comprises a comb for injection of electrophoresis medium into the cassette, as well as the corresponding method of injection.
- (b) Background of the Invention
- Electrophoresis is a well known separation technique that requires the application of electrical current at both poles of a cassette or plate to force samples through an electrophoretic medium that acts as a molecular sieve. The application of a difference of potential between the upper section and the lower section of the cassette assumes the creation of two areas sealed from each other. Because current is transmitted via two separate buffer reservoirs, it is necessary to apply a pressure or force on the cassette so that the seals properly operate. It is therefore imperative that the whole system, including the cassette, possess some rigidity.
- Conventional electrophoresis cassettes are made of two glass plates spaced apart with plastic spacers or tongues to create a space therebetween while ensuring that the sides of the assembly are properly sealed. Importantly, the spacers must not conduct electrical current. The assembly is generally maintained together with clamps, and it is often necessary to reinforce the seals with hot agar. When the gel is cast into the cassette, a comb element is introduced at one end of the assembly to create one or more reservoirs or wells thereafter wherein the sample(s) will be received later. The shape of the comb may comprise various numbers and sizes of reservoirs, depending on the application required and the size of the cassette. For example, a preparation gel necessitate less reservoirs, while an analytical gel will require more reservoirs and the width thereof will depend on the resolution desired.
- However, such assemblies have several drawbacks and limitations. The assembling operation requires dexterity and is a time-consuming operation, because it is done manually. The plates are conventionally made of glass, and thus must be handled with care. Further, they must be carefully cleaned to obtain good results. Finally, manipulation of acrylamide gel, a commonly used electrophoretic medium, represents a long-term danger for the health of operators since such gel is highly toxic.
- More recently, to simplify the assembling work of operators and reduce poisoning and manipulation hazards, pre-cast cassettes already containing the gel have been made available commercially. The cassettes comprise an acrylamide gel, and a comb is provided at one extremity thereof. However, the cost of these cassettes is prohibitive, and demolding thereof, for visualization of the results, is a delicate and complicated procedure. In addition, the comb is produced by injection molding, and is used to form the wells or reservoirs in the gel. They generally represent an important part of the total cost of the cassette.
- To be economically feasible and capable of supporting, without substantial bending, the mechanical forces applied thereon, cassettes containing pre-cast electrophoresis medium, must be rigid enough and made of a material economically sound and preferably recyclable, such as for example thermoplastic materials like polymethylmethacrylate (PMMA). However, conventionally, in order to be sufficiently rigid, the plates must be relatively thick. Two obvious problems therefore become apparent: a) the amount of thermoplastic material required is significant, thus increasing the cost, which is not suitable for a disposable device; and b) maintaining the gel at an appropriate operating temperature is complicated, because the thick walls of the thermoplastic material act as a dielectric material. Thicker plastic walls also affect the diffusion of the heat generated during the electrophoretic process, creating temperature gradients within the electrophoresis medium, and non-uniform migration of the samples analyzed.
- Conventional processes for filling the cassettes are generally standard, irrespective of the electrophoretic medium. Typically, a gel comprising a mixture of acrylamide and bis-acrylamide, a buffer like tris-borate ethylenediamine (EDTA), tris-acetate-EDTA, tris-glycine, tricine, and a polymerization initiator are injected or cast into the cassette. Some of these products are neurotoxic and/or irritant, and must therefore be handled with extreme care. A laboratory pipette or a pump can be used to fill the cassette from the top with the liquid medium. Once the cassette is filled, a comb closes the top of the cassette. The comb has a design such that it contains one or more teeth forming reservoirs in the gel wherein the sample will be placed later. After polymerization of the medium, the comb is removed, as well as a separator present in the lower portion of the cassette. The cassette is then placed in an electrophoresis apparatus wherein the lower and upper portions of the gel will be in contact with two independent buffer solutions. The samples are then introduced in the reservoirs, and current is applied to separate the various components of each sample. After completion of the separation, the medium is removed from the cassette for further processing, i.e., coloration, photograph and analysis.
- Again, such system and procedure have various major drawbacks and limitations. As stated above, manual filling of the cassette requires great care and dexterity, not to mention exposure of the operator to toxic chemicals. Further, undesirable bubbles often form during filling, and installation of the comb after filling may also create bubbles at the bottom of the teeth. Such air bubbles must be avoided at all times, since they interfere significantly with the samples migrating in the polymerized gel during the electrophoresis procedure.
- Pre-cast gels have been marketed recently, but have not been able to overcome other problems mentioned above for cassettes containing the same, such as prohibitive costs. One of the main reason is that the cassettes are obtained by injection molding, which is a costly and relatively slow process because of the significant amount of plastic required for injection, the cost of the plastic material itself, and the time necessary to allow complete cooling of the cassette thus obtained. In addition, because the cassettes are made of a thermoplastic material, gel polymerization is greatly affected and slowed down because the polymer absorbs free radicals generated by the chain reaction of the polymerization. As a result, the polymerized electrophoretic medium does not “stick” do the cassette inner surfaces. An expensive coating layer or overlay must therefore be applied on the thermoplastic material surfaces to minimize this problem and ensure proper polymerization quality and speed.
- The electrophoresis operation necessitates the application of a voltage across the gel that generates heat that must be somehow dissipated. During the heat dissipation process, if the temperature of the gel is not uniform, it causes distortion in the separated protein or polynucleic acid bands. Such heat is therefore a critical problem because it limits the rate at which gels can be run. Increasing temperatures reduces the resistance and increases current at a given voltage. Although the net effect is a shorter run, excessive temperature can lead to undesirable band broadening. It is therefore preferable to run at a higher voltage and a constant lower temperature.
- The present invention is concerned with an electrophoresis cassette comprising:
- first and second thermoformed surfaces comprising their edges in hermetic contact with each other and defining a volume therebetween for receiving an electrophoretic medium;
- either of first or second surface comprising at least one opened reservoir molded therein at one end; and
- either of first or second surface further comprising at least one aperture at an opposite end thereof.
- In a preferred embodiment, the cassette comprises a male member having a structure complementary to that of the at least one reservoir, the male member being adapted to be removably inserted therein and further comprising:
- an inlet and an outlet for injecting the electrophoretic medium therethrough and into the volume defined by the first and second surfaces; and
- at least one opening for discharge of the excess of the electrophoretic medium.
- In a second aspect of the invention, there is provided a comb for an electrophoretic cassette adapted to be removably inserted into the cassette and allowing injection therein of an electrophoretic medium and discharge of excess of electrophoretic medium therefrom, comprising at least one inlet and one outlet for injection of the electrophoretic medium into the cassette, the outlet extending longitudinally on a first side of at least one tooth.
- In a third aspect of the invention, there is provided an electrophoresis assembly comprising:
- a cassette as defined above;
- an electrophoretic medium comprised in the volume; and
- a support comprising a frame adapted to receive the cassette, the support further comprising a plurality of recesses forming corresponding channels between a cassette surface and the support, thus allowing passage of a coolant therein, whereby the temperature of the medium is substantially the same within the volume.
- In a fourth aspect of the invention, there is provided a method for casting an electrophoretic medium into a cassette, comprising the steps of:
- sealing any aperture present in the cassette;
- injecting the electrophoretic medium into the inlet of the comb defined above, the medium penetrating into the volume through the comb; and
- continuing injection of the medium until all the volume is filled and an excess thereof is discharged from at least one opening of the comb.
- Finally, in a fifth aspect of the invention, there is provided a composition for use as an electrophoretic medium comprising:
- an electrophoretic gel;
- a polymerization initiator;
- a buffer; and
- an adhesive agent allowing adhesion of polymerized electrophoretic medium onto a surface of an thermoplastic electrophoretic cassette.
- FIG. 1 illustrates a cassette and corresponding support plate in accordance with the present invention;
- FIG. 2 illustrates the details of a comb developed in accordance with the present invention; and
- FIG. 3 illustrates the support plate developed for supporting the present cassette.
- The invention relates to the field of electrophoresis, and more particularly to a cassette suitable therefor. It is to be assumed that the gel used as the electrophoretic medium is preferably an acrylamide (or polyacrylamide) gel, whether cross-linked or not. However, other conventional and well known electrophoretic media such as agarose gel or starch gel, can be used. Polyacrylamide gel is particularly preferred because it is transparent, electrically neutral, and can be made in various pore sizes. Other co-monomers well known in the field include N,N′-methylenebisacrylamide, N,N-bisacrylylcystamine, N,N′-(1,2-dihydroxyethylene)bisacrylamide, N,N′-diallyl-tartardiamide, and the like.
- The drawings provided herewith are for the sole purpose of illustrating preferred embodiments of the invention, and shall not be considered as limiting the scope thereof.
- Referring to drawings, FIG. 1 illustrates a
cassette assembly 10 and asupport 12 therefor.Cassette 10 comprises atop plate 14 and areservoir plate 16 each of a substantially square shape and having their four edges sealed, either with anadhesive layer 11 inserted therebetween, or with the help of any other compatible sealing means such as glue, ultrasonic welding, tape etc. The structure oflayer 11 is complementary with that of bothplates -
Plates plates least reservoir plate 16 should be transparent, but it is preferred that bothplates -
Reservoir plate 16 comprises a series of reservoirs 18 for receiving a corresponding series ofteeth 20 ofcomb 22.Top plate 14 has a complementary structure, i.e., a series ofopenings 24, that allow the passage therethrough of the plurality ofteeth 20 for engagement into reservoirs 18.Reservoir plate 16 further comprises a series ofslots 26 aligned with the series of reservoirs 18, and of substantially the same width. During filling, shipping and handling operations, these slots are sealed with a removable sealing strip 28 that will be removed before placingcassette 10 in the electrophoresis device. In an alternate embodiment, it has been found that the series ofslots 26 can be replaced with slots having a smaller width but being present in a greater number, i.e, preferably twice the number ofslots 26, with the same end result. -
Comb 22 comprises an aperture orinlet 30 extending therethrough substantially perpendicularly to its longitudinal section, and aligned with atooth 32, the latter comprising alongitudinal recess 34 shown in phantom lines in FIG. 2 and serving as an outlet. After engagement ofteeth 20 into the series of reservoirs 18, an electrophoretic medium is injected intocassette 10 throughaperture 30 andrecess 34, as indicated by arrow 36. The flow of electrophoretic medium insidecassette 10 is also indicated byarrows cassette 10, as well as minimizing air bubbles, a slight excess of electrophoretic medium must be injected. Such excess is discharged out ofcassette 10 through a longitudinal recess 38 provided in each tooth 18. The flow of discharge is indicated byarrow 44. Recess 38 is located on the side of atooth 20 that is opposite to the side oftooth 32 comprisingrecess 34. Eachtooth 20 further comprises a pair ofgrooves comb 22 upon removal thereof after completion of polymerization of the electrophoretic medium, although they may also be useful for discharge of excess of gel.Grooves comb 22 from the gel, thus leaving a lower surface of reservoir 18 containing the medium substantially similar and even in each reservoir 18. - During the electrophoretic medium casting process, the medium is poured into
cassette 10 through opening 30 ofcomb 22, and allowed to solidify. Preferably, the cassette is held in a manner such thatplates Plates cassette 10. This method therefore substantially eliminates air bubbles fromcassette 10. Once polymerization is complete,cassette 10 is stored appropriately in a conventional manner. -
Comb 22 is preferably removed only minutes prior to the use of the cassette, or immediately after complete polymerization of the gel, prior to storage, if the reservoirs 18 are well preserved from dehydration. At that point, it is slowly pulled out of the cassette, and each reservoir 18 is thereafter filled with an appropriate volume of a sample to be electrophoresed. - It is well known that in operation, the temperature of the electrophoresis gel increases. It is also well known that the temperature will be higher towards the middle of the cassette than on the sides thereof. As a result, the migration front of the products to be separated is altered, and erroneous interpretation might occur. A novel support plate has therefore been developed to overcome these problems, as well as for providing a proper profile maintenance, i.e., sufficient rigidity of the thin walls of the cassette, and facilitating installation of the cassette into an operational position in a conventional electrophoresis apparatus.
-
Support plate 12 comprises aframe 46 adapted to receive thereincassette 10, and comprising asurface 48 with a plurality oflongitudinal recesses 50, which can be of any shape and size.Openings slots 26. Whencassette 10 is placed ontosupport 12, it lies directly ontoridges 56 ofplate 12, thus forming a series of channels betweenrecesses 50 and a surface ofcassette 10 for circulation of the buffer solution therein (flow indicated by arrows 51), and thus helping dispersing heat generated within the cassette. As illustrated, eachrecess 50 is preferably aligned with a reservoir 18 and aslot 26, to ensure that the temperature of the migrating product and the gel is substantially the same, whether the reservoir is near the side or the middle of the cassette. It has however been found that such alignment is not mandatory. The critical element is that some buffer solution is allowed to circulate between the support plate and the cassette to “extract” heat from the latter.Support 12 can be made of any suitably rigid material, but is preferably made of a heat conducting material, so that heat is also extracted fromridges 56 that are in direct contact with the surface ofcassette 10 lying thereon, and dispersed within the structure of the support.Cassette 10 can be maintained in place inplate 12 with the help of a couple ofretainer plates 58. - With respect to the problem of interference of the polymerization process caused by the thermoplastic material of the cassette, it has been found that by combining a powerful initiator generating more free radicals with an appropriate “sticking” agent in the gel composition, there is no longer a need to apply a costly protective layer over the thermoplastic surfaces. Examples of such initiators include ammonium persulfate and N,N,N,N-tetramethylethylenediamine (TEMED); 4-dimethylaminopropionitrile; 1-hydroxycyclohexyl phenyl ketone; 2,2-diethoxy-acetophenone; 2,2-dimethoxy-2-phenylacetophenone; 2′,4′-dimethoxy-acetophenone; 2-hydroxy-2-methyl-1-propiophenone; 2-hydroxy-2-methyl-1-phenylpropan-1-one, and mixtures thereof. These strong initiators allow a substantially complete polymerization of the gel. However, the resulting polymerized gel does not stick to the plastic surface, which is critical, particularly in view of the fact that the cassette structure is relatively flexible. Detachment or unsticking of the polymerized electrophoretic medium from the cassette inner surfaces may lead to the introduction of undesirable air bubbles between the plastic surface and the gel, and may also cause irregularities in the medium structure, thus severely impairing the efficiency of the cassette. Surprisingly, it has been found that by adding to the gel composition a small amount of an adhesive compound is sufficient to allow the gel to adequately stick onto the plastic surface. The adhesive compound preferably corresponds to that used for coating the inner surfaces of currently available thermoplastic cassettes for the same purpose. However, the costs associated with the processing and coating of such a layer on the inner surfaces of the cassette are significant. On the other hand, in the present invention, all one has to do is to add a sufficient amount of the said adhesive compound into the gel composition to be injected into the cassette to achieve the same result. Not only is the procedure more simple, but the amount of adhesive compound required is smaller. Suitable adhesive compounds include polysilazanes or tetra-substituted silicon derivatives. The substituents can be the same or different, and include a straight or branched alkyl, alkoxy, ketone, ester or amide each comprising from 1 to 8 carbon atoms, or an amino, halogen, cyano or hydroxy. Preferred adhesives are alkyl alkoxy silane derivatives. Most preferred adhesives include Silane A-174, methacryloxytrimethoxysilylpropane, 3-(trimethoxysilyl)propyl methacrylate, 3-methacryloxypropyltrimethoxysilane, MEMO, DYNASYLAN MEMO, and γ-methacryloxypropyltrimethoxysilane.
- The thickness of
plates - While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications, and this application is intended to cover any variations, uses or adaptations of the invention following, in general, the principles of the invention, and including such departures from the present description as come within known or customary practice within the art to which the invention pertains, and as may be applied to the essential features hereinbefore set forth, and as follows in the scope of the appended claims.
Claims (2)
1. A composition for use as an electrophoretic medium comprising:
an electrophoretic gel;
a polymerization initiator;
a buffer; and
an adhesive agent allowing adhesion of polymerized electrophoretic medium onto a surface of an thermoplastic electrophoretic cassette.
2. A composition according to claim 1 wherein the gel comprises acrylamide gel, optionally cross-linked, agarose gel or starch gel; the polymerization initiator comprises ammonium persulfate and N,N,N,N-tetramethylethylenediamine (TEMED); 4-dimethylaminopropionitrile; 1-hydroxycyclohexyl phenyl ketone; 2,2-diethoxy-acetophenone; 2,2-dimethoxy-2-phenylacetophenone; 2′,4′-dimethoxyacetophenone; 2-hydroxy-2-methyl-1-propiophenone; 2-hydroxy-2-methyl-1-phenylpropan-1-one, and mixtures thereof; the buffer comprises tris-borate EDTA, tris-acetate-EDTA, tris-glycine, tricine, or mixtures thereof; and the adhesive agent comprises at least one tetra-substituted silicon derivative wherein the substituents are the same or different and comprise a straight or branched alkyl, alkoxy, ketone, ester or amide each comprising from 1 to 8 carbon atoms, amino, halogen, cyano or hydroxy.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/078,401 US20020079222A1 (en) | 1999-09-01 | 2002-02-21 | Composition for use as an electrophoretic medium |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US09/387,758 US6379519B1 (en) | 1999-09-01 | 1999-09-01 | Disposable thermoformed electrophoresis cassette |
US10/078,401 US20020079222A1 (en) | 1999-09-01 | 2002-02-21 | Composition for use as an electrophoretic medium |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/387,758 Division US6379519B1 (en) | 1999-09-01 | 1999-09-01 | Disposable thermoformed electrophoresis cassette |
Publications (1)
Publication Number | Publication Date |
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US20020079222A1 true US20020079222A1 (en) | 2002-06-27 |
Family
ID=23531266
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09/387,758 Expired - Fee Related US6379519B1 (en) | 1999-09-01 | 1999-09-01 | Disposable thermoformed electrophoresis cassette |
US10/078,401 Abandoned US20020079222A1 (en) | 1999-09-01 | 2002-02-21 | Composition for use as an electrophoretic medium |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
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US09/387,758 Expired - Fee Related US6379519B1 (en) | 1999-09-01 | 1999-09-01 | Disposable thermoformed electrophoresis cassette |
Country Status (10)
Country | Link |
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US (2) | US6379519B1 (en) |
EP (2) | EP1208371B1 (en) |
JP (1) | JP2003508751A (en) |
AT (1) | ATE243317T1 (en) |
AU (2) | AU771464B2 (en) |
CA (1) | CA2381962C (en) |
DE (1) | DE60003437T2 (en) |
ES (1) | ES2202159T3 (en) |
MX (1) | MXPA02002328A (en) |
WO (1) | WO2001016589A1 (en) |
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Also Published As
Publication number | Publication date |
---|---|
JP2003508751A (en) | 2003-03-04 |
DE60003437T2 (en) | 2004-04-29 |
ATE243317T1 (en) | 2003-07-15 |
US6379519B1 (en) | 2002-04-30 |
CA2381962A1 (en) | 2001-03-08 |
EP1231466A2 (en) | 2002-08-14 |
AU2004200643A1 (en) | 2004-03-11 |
EP1208371A1 (en) | 2002-05-29 |
CA2381962C (en) | 2007-11-20 |
AU771464B2 (en) | 2004-03-25 |
EP1231466A3 (en) | 2004-01-21 |
MXPA02002328A (en) | 2004-07-16 |
WO2001016589A1 (en) | 2001-03-08 |
ES2202159T3 (en) | 2004-04-01 |
AU6678800A (en) | 2001-03-26 |
DE60003437D1 (en) | 2003-07-24 |
EP1208371B1 (en) | 2003-06-18 |
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