Programmable electrophoresis with integrated and multiplexed control

• US 5,185,071 A
• Filed: 10/30/1990
• Issued: 02/09/1993
• Est. Priority Date: 10/30/1990
• Status: Expired due to Fees

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Dunn, Frederick J., Serwer, Philip
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US07/605,796
• Time in Patent Office: 833 Days
• Field of Search: 204/299 R, 204/182.8, 204/182.1
• US Class Current: 204/457
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis

Abstract

A user programmable device for horizontal gel eletrophoresis is provided. This device can both accurately and programmably control: (a) the angle between electrical field and gel by use of a rotatable gel bed, (b) magnitude of the electrical field, and (c) temperature. The present invention is programmable by a microprocessor-based contol board which sends control signals to: (a) a motor attached to the rotatable gel (b) the source of elecrical potential and (c) Peltier devices attached to the sides of buffer chambers horizontally displaced from the rotatable gel bed. The present invention is particularly appropriate for lowering cost, lowering footprint, reducing heat dissipated, maintaining close tolerances of electric field gradients and temperature, and exploring and using thus far untried modes of electrical field variation. It is thereby suitable for analyzing large linear DNA, open circular DNA and/or DNA-protein complexes. Control of several disks, power supplies and temperature cells by one control board (multiplexing) is an option.

Claims

1. An apparatus for programmably controlling an electrophoretic device, comprising:

a planar rotatable disk for supporting a horizontal gel containing at least on DNA particle;

programmable rotating means connected to said disk for rotating said disk;

at least two buffer holding tanks horizontally displaced from said rotatable gel bed, said tanks open at the top to provide electrically conductive buffer solution between said tanks and over said gel;

programmable controlling means connected to said holding tanks for controlling the magnitude and polarity of the electric field placed between said tanks; and

programmable Peltier means connected to said holding tanks for regulating the temperature of said buffer solution within said tanks and over said gel.

2. The apparatus as recited in claim 1, further comprising a means for containing a thin layer of said buffer solution upon the upper surface of said gel.

3. The apparatus as recited in claim 2, wherein said buffer solution is a phosphate-based solution.

4. The apparatus as recited in claim 1, wherein said rotating means comprises:

a DC stepping motor mechanically coupled to said support;

a home finder disk mechanically coupled to said support; and

a control board electrically connected to said motor and said disk for controlling the speed, direction and duration of movement of the motor.

5. The apparatus as recited in claim 1, wherein said controlling means comprises:

an electrode placed in each holding tank for receiving electrical inputs; and

a control board electrically connected to each electrode for controlling magnitude, polarity and duration of electrical energy supplied to each said electrode.

6. The apparatus as recited in claim 1, wherein said Peltier means comprises:

each said holding tank having an internal and external surface, said internal surface configured for containing said solution;

a thermally conductive sheet having an internal and external surface, whereby said internal surface of said sheet is placed on the external surface of each said holding tank covering a hole in each said holding tank;

whereby said internal surface of said plate is attached to the external surface of each said thermally conductive sheet;

a Peltier cell having an internal and external surface, whereby said internal surface of said cell is attached to the external surface of each said plate;

an aluminum heat sink having a planar inner surface attached to the external surface of said cell; and

a control board electrically connected to each cell for inputting electrical input to the cell which, in response to the electrical input, the cell transfers heat to said heat sink.

7. The apparatus as recited in claim 1, wherein said Peltier means comprises:

each said holding tank having an internal and external surface, said internal surface configured for containing said solution;

a thermally conductive sheet having an internal and external surface, whereby said internal surface of said sheet is placed on the external surface of each said holding tank covering a hole in each said holding tank;

a Peltier cell having an internal and external surface, whereby said internal surface of said cell is attached to the external surface of each said sheet;

an aluminum heat sink having a planar inner surface attached to the external surface of said cell; and

a control board electrically connected to each cell for inputting electrical input to the cell which, in response to the electrical input, the cell transfers heat to said heat sink.

8. The apparatus as recited in claim 1, wherein said Peltier means comprises:

each said holding tank having an internal and external surface, said internal surface configured for containing said solution;

a Peltier cell having an internal and external surface, whereby said internal surface of said cell is attached to the external surface of each said holding tank covering a hole in each said holding tank;

an aluminum heat sink having a planar inner surface attached to the external surface of said cell; and

a control board electrically connected to each cell for inputting electrical input to the cell which, in response to the electrical input, the cell transfers heat to said heat sink.

9. The apparatus as recited in claim 4, 5 or 8, wherein said control board is a microprocessor and memory-based controller with programmable pulse generator.

10. A method for multiplexing control of both electrical field and temperature during electrophoresis, comprising the steps of:

placing particles onto a rotatable gel bed;

containing a buffer solution over said bed in communication with said particles;

applying a variable electric field to said bed at varying angles of incidence in response to programmed input; and

drawing thermal energy from said solution in response to programmed electrical input.

11. The method as recited in claim 10, wherein the applying step comprises rotating said particles and buffer solution in a horizontal plane.

12. The method as recited in claim 10, wherein said drawing step comprises passing current through a Peltier device.

13. The method as recited in claim 10, further comprising multiplexing control of at least two gel beds, each bed containing said solution for receiving independently applied electrical field and independently drawn said thermal energy.

14. An apparatus for programmably controlling electrophoresis, comprising:

a substantially planar rotatable support member for supporting a medium adapted to hold at least one sample to be electrophoresis separated;

means for containing a buffer solution upon an upper surface of said rotatable support;

first programmable means for controlling an orientation of said rotatable support and for controlling a magnitude and polarity of an electric field acting upon said medium;

a thermoelectric chilling device for controllably chilling said buffer solution; and

control means for controlling said programmable means and said thermoelectric chilling device, independent of one another.

15. An apparatus for programmably controlling electrophoresis comprising:

a substantially planar rotatable support member for supporting a medium adapted to hold at least one sample to be electrophoresis separated;

means for containing a buffer solution upon an upper surface of said rotatable support;

programmable means for controlling a magnitude and polarity of an electric field acting upon said medium;

a controllable stepping motor for controllably rotating said rotatable support member;

a Peltier device for controllably chilling said buffer solution; and

control means for controlling said programmable means said Peltier device and said stepping motor, independent of one another.