Horizontal electrophoresis separation device without seal and method of extracting gel without opening cassette

Patent metadata

Field	Value
Publication number	US-2016299099-A1
Application number	US-201514968400-A
Country	US
Kind code	A1
Filing date	Dec 14, 2015
Priority date	Dec 16, 2014
Publication date	Oct 13, 2016
Grant date	—

Abstract

Horizontal gel electrophoresis devices and methods of extracting a gel from a gel housing are provided. The devices allow for extraction of gels without having to open the housing comprising the gel. The devices include gel cassettes that simplify the automation of gel electrophoresis and electroblotting of proteins. Also provided are devices that allow for horizontal gel electrophoresis without the need to from a liquid seal between the buffer reservoir and the gel housing. Also described are methods for extracting a gel from a housing without opening the housing.

First claim

1 . A device for horizontal electrophoresis, the device comprising:
a first and second chamber for holding buffer, the first and second chamber separated from each other and each comprising a floor, wherein the floor of the first and second chamber each comprises one or more opening providing fluid communication between the first and second chambers and one or more separation channel; the separation channel(s) residing below the floor of the first and second chambers; and a cathode disposed within the first chamber and an anode disposed within the second chamber.

2 . The device of claim 1 , wherein the openings in the floor of at least one of the first and second chambers form sample wells that protrude above the surface of the floor such that a volume of sample can be contained in each well.

3 . The device of claim 1 , wherein the one or more separation channel(s) comprise an electrophoresis gel.

4 . The device of claim 3 , wherein the separation channels each have a first and second end, the first end of said separation channels abutting an extrusion member capable of sliding to extrude the electrophoresis gel in the separation channel out of the second end of the separation channel.

5 . The device of claim 4 , further comprising a track aligned with each separation channel to receive a gel extruded by the extrusion member from the separation channel.

6 . The device of claim 5 , further comprising a first transfer electrode positioned above the track and a second transfer electrode positioned below the track, wherein the track is part of a solid support comprising pores and a medium for conducting ions.

7 . (canceled)

8 . (canceled)

9 . The device of claim 1 , wherein the floor of the first or second chamber, and/or the bottom and/or the top of the separation channel comprises a transparent material.

10 . The device of claim 1 , wherein the device comprises a plurality of separation channels.

11 . (canceled)

12 . (canceled)

13 . The device of claim 1 , wherein the first and second chamber are separated from each other by a dividing wall, or the first and second chamber are horizontally separated from each other such that at least a portion of the separation channel is visible between the first and second chambers.

14 . (canceled)

15 . (canceled)

16 . The device of claim 1 , wherein the first and/or second chamber further comprises an inlet and an outlet configured to circulate buffer by way of a pump that flows buffer from the inlet to the outlet.

17 . A device for horizontal electrophoresis, the device comprising:
a first chamber for holding buffer comprising a floor, wherein the floor of the first chamber comprises one or more opening providing fluid communication between the first chamber and one or more separation channel; the separation channel(s) residing below the floor of the first chamber; and a cathode disposed within the first chamber.

18 . (canceled)

19 . The device of claim 17 , wherein the one or more separation channel(s) comprise an electrophoresis gel, wherein the separation channels each have a first and second end, and further comprising an extrusion member in slidable contact with the electrophoresis gel at the first end of the separation channel, wherein the extrusion member is configured to slidably extrude the electrophoresis gel in the separation channel out of the second end of the separation channel.

20 - 24 . (canceled)

25 . A device for horizontal electrophoresis, the device comprising,
a first and second chamber for holding buffer, each of said chambers composed of a floor, two side walls, an outer wall, an inner wall, and a dividing wall, said dividing wall separating an inner portion and an outer portion of the chamber, said dividing wall having one or more dividing wall openings to allow for buffer flow between said inner and outer portions, each of the chambers configured to circulate buffer between said inner and outer portions, and each chamber comprising an electrode; disposed between, and attached to, the inner walls of said first and second chambers, one or more platform for receiving a horizontal gel housing, wherein the dividing wall openings horizontally align with a gel in the gel housing, if present, such that buffer in the chambers can contact the gel, if present, and pass from the inner portions to the outer portions.

26 . (canceled)

27 . (canceled)

28 . The device of claim 25 , wherein the inner and outer portions of the chambers comprise one of an outlet and an inlet and the chambers are configured to circulate buffer between said inner and outer portions by way of a pump that flows buffer from the inlet to the outlet with buffer returning via the dividing wall openings, wherein said inlet is in said inner portion and said outlet is in said outer portion.

29 . (canceled)

30 . The device of claim 25 , further comprising the horizontal gel housing, wherein the horizontal gel housing comprises a top and a bottom, wherein the bottom rests on the one or more platform, and the top comprises one or more barrier to prevent buffer from flowing from one chamber to another of said chambers.

31 . (canceled)

32 . (canceled)

33 . A method of extracting an electrophoresis gel without opening a housing, the method comprising:
providing a housing containing the gel, the housing comprising open first and second ends, wherein the gel is exposed at the first end and second end; contacting the exposed gel at the first end with an extrusion member with sufficient force to push the gel out of the second end of the housing, thereby extracting the gel from the housing.

34 . (canceled)

35 . (canceled)

36 . The method of claim 33 , wherein the housing comprises one or more gel electrophoresis channels, and the gel is contacted at a first end of the channel with the extrusion member with sufficient force to push the gel out of a second end of the channel.

37 . (canceled)

38 . (canceled)

39 . A method for analyzing an analyte following gel electrophoresis, the method comprising:
extruding a gel comprising analytes separated by size or charge from a separation channel of the device of claim 1 onto a track aligned with the separation channel; contacting the extruded gel with an electric field between first and second transfer electrodes such that the analytes are electrophoretically transferred to a solid support; analyzing the analyte.

40 . (canceled)

41 . The method of claim 39 , wherein the gel is extruded by contacting the gel with an extrusion member with sufficient force to push the gel out of the separation channel onto the track, or the gel is extruded by vibrating the device of claim 1 under conditions suitable for extruding the gel from the separation channel onto the track.

42 . (canceled)

43 . (canceled)

44 . A system for aligning images of a gel, comprising:
a device of claims 1 comprising an electrophoresis gel comprising a dye band; a detector capable of detecting the dye band; a computer configured with executable instructions for aligning a first image of the gel and a second image of the gel, wherein the firs

Assignees

• Bio Rad Laboratories Inc

Inventors

• Uri Yochanan
• Issman Uri
• Kochetkov Igor
• Barak Itay
• Aharon Shai
• Nimri Shai

Classifications

• G01N27/44743

  Introducing samples · CPC title

• G01N27/44756 Primary

  Apparatus specially adapted therefor · CPC title

• G01N27/44721

  by optical means · CPC title

• G01N27/44726

  using specific dyes, markers or binding molecules · CPC title

• G01N33/54366

  Apparatus specially adapted for solid-phase testing · CPC title