Claims
- 1. A method for making an electrophoresis gel, comprising the steps of:
- (a) applying a plurality of solid particles to a surface of a first substrate;
- (b) forming a gel holder comprising the first substrate, and a second substrate by partially sealing the first substrate to the second substrate such that the plurality of solid particles is disposed between the first and second substrate, and such that a gel compartment having a thickness 250 microns or less is formed between the first substrate and the second substrate, said gel compartment having an opening for the introduction of gel;
- (c) filling the gel compartment with an unpolymerized gel; and
- (d) polymerizing the gel in the gel compartment, wherein the solid particles have a size substantially equal to the thickness of the gel compartment.
- 2. A method according to claim 1, wherein the solid particles are applied to the substrate as a mixture with a powdered, low-melting glass further comprising the steps of
- heating the substrate having the solid particles and the powdered, low-melting glass thereon to melt the low-melting glass, and
- cooling the heated substrate to resolidify the low melting glass, whereby the solid particles are bonded to the surface of the substrate by the resolidified glass.
- 3. A method according to claim 1, wherein the first substrate is sealed to the second substrate with an adhesive.
- 4. A method according to claim 3, wherein the adhesive is applied by screen printing.
- 5. A method according to claim 3, wherein at least a portion of the solid particles are dispersed within the adhesive.
- 6. A method according to claim 5, wherein the adhesive is an acrylate adhesive.
- 7. A method according to claim 1, wherein the separation between the substrates is from 25 to 100 microns.
- 8. A method according to claim 1, wherein the gel is polyacrylamide.
- 9. A method according to claim 1, wherein the gel is separated into two or more lanes.
- 10. A method according to claim 9, wherein the lanes are separated by lines of adhesive containing at least a portion of the solid particles.
- 11. A method according to claim 1, wherein at least a portion of the solid particles are disposed on a surface of the top substrate or the bottom substrate prior to the formation of the gel compartment, whereby the plurality of particles are disposed within the gel compartment of the microgel.
- 12. A method according to claim 11, wherein the solid particles are disposed on the top substrate or the bottom substrate by application of a slurry of particles in a volatile liquid which is evaporated prior to the formation of the gel compartment.
- 13. A method according to claim 1, wherein the gel is photopolymerized.
- 14. A method according to claim 1, further comprising the step of cutting sample introduction and electrode holes in the top substrate prior to its assembly into the gel holder.
- 15. A method according to claim 14, wherein the sample introduction and electrode holes are sealed over by a removable adhesive strip prior to the filling of the gel compartment.
- 16. A method according to claim 15, wherein the removable adhesive strip permits transmission of UV light.
- 17. A method according to claim 11, further comprising the step of cutting a substantially rectangular sample introduction hole into the top substrate prior to its assembly into a gel holder, said sample introduction hole being formed near a first end top substrate such that sample loaded through the sample introduction hole will migrate towards a second, opposite end of the top substrate.
- 18. A method according to claim 17, wherein the sample introduction hole has a long edge closest to the second end of the substrate, and said long edge is beveled such that the sample introduction hole has larger dimensions on a first surface than on an opposite second surface, and wherein the gel holder is assembled such that the first surface is on the inside of the gel holder.
- 19. A method according to claim 18, further comprising the step of placing an insert into the sample introduction hole prior to polymerization of the gel, wherein the insert is beveled to match the bevel of the sample introduction hole such that a trough is formed between the beveled edge of the top substrate and the gel into which sample may be loaded.
- 20. A method for analyzing a mixture of labeled oligonucleotide fragments of different lengths comprising:
- (a) loading the mixture onto an origination site on a microgel,
- (b) applying a potential to the electrophoresis gel creating an electric field strength of 100 V/cm or greater, whereby a current flows through the electrophoresis gel and transports the oligonucleotide fragments in the mixture away from the origination site and through a monitoring site remote from the origination site on the gel; and
- (c) detecting oligonucleotides passing the monitoring point, wherein the microgel comprises
- a top substrate and a bottom substrate sealed to one another to form a gel compartment having a thickness of from 250 microns or less,
- a plurality of solid particles having a mean diameter substantially equal to the thickness of the gel compartment, said plurality of solid particles being disposed within the gel compartment, and
- an electrophoresis gel disposed within the gel compartment.
- 21. A method according to claim 20, wherein the electrophoresis gel is polyacrylamide.
- 22. A method according to claim 20, wherein at least a portion of the solid particles are dispersed in an adhesive which seals the top substrate to the bottom substrate.
- 23. A method according to claim 22, wherein the adhesive is an acrylate adhesive.
- 24. A method according to claim 20, wherein at least a portion of the solid particles are bonded to at least one of the substrates using a low-melting glass.
- 25. A method according to claim 20, wherein the oligonucleotides fragments are fluorescently-labeled, and are detected as they pass the monitoring point using a fluorescence detector.
- 26. A method according to claim 25, wherein the fluorescently-labeled oligonucleotides are detected using an argon ion laser.
- 27. A method according to claim 20, wherein an electric field strength of greater than 150 V/cm is created.
- 28. A method according to claim 20, wherein an electric field strength of from 250 to 400 V/cm is created.
- 29. A method according to claim 20, wherein at least a portion of the solid particles are dispersed within the gel in the gel compartment.
- 30. A loading insert for use in loading a sample onto an electrophoresis gel, comprising
- (a) a substantially rectangular planar base portion having two long edges and two short edges and upper and lower faces;
- (b) a plurality of fingers extending from a first long edge of the base portion in a plane with the upper and lower faces of the base portion, each of said fingers having a width of from 0.5 to 3 mm; said fingers being evenly spaced at intervals of 2 to 7 mm in a region along the first long edge of the base portion starting and ending at a point from 2 to 4 mm from the adjacent short edge of the base portion; and each of said fingers having a tip portion at the distal end thereof, said tip portion being cut with a 30.degree.-60.degree. bevel relative to the plane of the base portion such that the lower surface of each finger is longer than the upper surface.
- 31. A loading insert according to claim 30, wherein the fingers each have a bevel of 45.degree. at the distal end thereof.
- 32. A loading insert according to claim 30, wherein lower surface of the loading insert has a plurality of grooves formed therein, said grooves being parallel to the fingers and extending across the entire loading insert.
- 33. A filling insert for use in filling an electrophoresis gel holder comprising a substantially rectangular planar body member having two long edges and two short edges, and upper and lower faces, wherein
- a first of said long edges is beveled at an angle of from 30 to 60 degrees relative to the plane of the body member such that the lower surface of the body member is larger than the upper surface, and
- a plurality of grooves are formed in the in the lower surface of the body member, said grooves running parallel to the short edges and extending from the one long edge to the other.
Parent Case Info
This application is a continuation-in-part and divisional of U.S. patent application Ser. No. 332,577 filed May 1, 1994, which is now U.S. Pat. No. 5,627,022.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
102e Date |
371c Date |
PCT/US95/14531 |
10/31/1995 |
|
|
5/1/1997 |
5/1/1997 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO96/13717 |
5/9/1996 |
|
|
US Referenced Citations (24)
Foreign Referenced Citations (3)
Number |
Date |
Country |
2273783 |
Jun 1994 |
GBX |
9300986 |
Jan 1993 |
WOX |
9300583 |
Jan 1993 |
WOX |
Non-Patent Literature Citations (2)
Entry |
Ansorge et al., "Thermally Stabilized Very Thin (0.02-0.3 mm) Polyacrylamide Gels for Electrophoresis", J Chromatography 202: 45-53 (1980). |
Maurer et al., "Polyacrylamide Gel Electrophoresis on Micro Slabs" Anal Biochem. 46: 19-32 (1972). |
Continuation in Parts (1)
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Number |
Date |
Country |
Parent |
332577 |
Nov 1994 |
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