Apparatus and method for a continuous rapid thermal cycle system

• US 8,293,471 B2
• Filed: 01/28/2005
• Issued: 10/23/2012
• Est. Priority Date: 01/28/2004
• Status: Active Grant

• Current Assignee: Marshall University Research Corporation
• Original Assignee: Marshall University Research Corporation
• Inventors: Gregg, Derek A., Murray, Elizabeth E., Norton, Michael L., Swick, Justin T., Tesser, Herbert
• Primary Examiner(s): Bowers, Nathan
• Application Number:
  US11/045,526
• Publication Number:
  US 20090325234A1
• Time in Patent Office: 2,825 Days
• Field of Search: 435/809, 435/303.1, 435/6.12, 435/287.3, 435/304.1, 435/91.2, 435/91.1
• US Class Current: 435/6.12
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/0841 Drums
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1883 using thermal insulation
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

A thermal cycle system and method suitable for mass production of DNA comprising a temperature control body having at least two sectors. Each sector has at least one heater, cooler, or other means for changing temperature. A path traverses the sectors in a cyclical fashion. In use, a piece of tubing or other means for conveying is placed along the path and a reaction mixture is pumped or otherwise moved along the path such that the reaction mixture is repetitively heated or cooled to varying temperatures as the reaction mixture cyclically traverses the sectors. The reaction mixture thereby reacts to form a product. In particular, polymerase chain reaction reactants may continuously be pumped through the tubing to amplify DNA. The temperature control body is preferably a single aluminum cylinder with a grooved channel circling around its exterior surface, and preferably has wedge-shaped or pie-shaped sectors separated by a thermal barrier.

Claims

1. A method for the facilitation of a chemical reaction requiring cyclical temperature changes for production of a product, comprising:

activating one or more means for changing temperature on a thermal cycle system, wherein said thermal cycle system comprises

a temperature control body comprising an exterior surface, at least two sectors forming a portion of said exterior surface, and a path cyclically passing through said sectors, wherein each sector comprises at least one of said means for changing temperature and is an independent temperature sink,

a tubing for conveying a fluid having a length, a first end, and a second end, said tubing further defining a volume and said tubing extending along said path, and

a means for moving in communication with said tubing, wherein said means for moving is adapted for moving said fluid through said tubing;

introducing a fluid, which is a substantially homogeneous temperature-dependent reaction mixture, into said tubing;

activating said means for moving such that said reaction mixture moves through said tubing, and such that said reaction mixture reacts to form a product;

continuously replenishing said reaction mixture at the second end of said tubing while said means for moving is activated so that said reaction mixture is continuously conveyed through the length of said tubing, so that said reaction mixture fills the entire volume of said tubing along the length of said tubing from the second end to the first end, and so that said reaction mixture is conveyed without impediment from the second end to the first end; and

collecting a continual supply of said product at the first end of said tubing so long as said means for moving is activated.

2. The method of claim 1, wherein said chemical reaction is a polymerase chain reaction.

3. The method of claim 2, wherein said substantially homogeneous temperature-dependent reaction mixture contains a non-ionic surfactant.

4. The method of claim 1, wherein said path is a grooved channel on said exterior surface of said temperature control body.

5. A method for the facilitation of a chemical reaction requiring cyclical temperature changes for production of a product, comprising:

controlling temperatures of at least twelve sectors so as to achieve a target temperature for each sector, wherein each sector is an independent temperature sink substantially made of a solid material and constitutes a respective portion of a single temperature control body;

conveying a temperature-dependent reaction mixture along a path through a tubing having a length, a first end, and a second end, said tubing further defining a volume, with the path passing through the at least twelve sectors repeatedly for several consecutive cycles, wherein for each cycle, the path passes once through a width of a first sector, and passes once through a width of one or more successive sectors, before returning to the first sector;

continuously replenishing said reaction mixture into the second end of said tubing such that said reaction mixture is continuously conveyed through said tubing, such that said reaction mixture fills the entire volume of said tubing along the entire length of said tubing from the second end to the first end, and such that said reaction mixture is conveyed without impediment from the second end to the first end; and

collecting a continual supply of said product at the first end of said tubing.

6. The method of claim 5, wherein the step of controlling temperatures includes:

setting the target temperature or temperature gradient range for each sector;

monitoring the temperature of each sector; and

adjusting the temperature of each sector to achieve and maintain its respective target temperature.

7. The method of claim 6, wherein the step of setting the target temperature or temperature gradient range for each sector includes setting the equivalent target temperature for at least two successive sectors.

8. The method of claim 5, wherein the single temperature control body has an exterior surface, each sector forming a portion of the exterior surface, wherein the path is a grooved channel on the exterior surface.

9. The method of claim 5, wherein the path is a channel formed internally within the temperature control body so as to pass internally through the sectors.

10. The method of claim 5, wherein the temperature control body is a cylinder having a circumference, wherein each sector is wedge-shaped, and wherein the path is a channel that spirals around the circumference of the cylinder.

11. The method of claim 5, wherein the width of each sector is substantially equivalent in size.

12. The method of claim 5, wherein the shape of said temperature control body is a three-dimensional shape of a geometrical form selected from the group consisting of a polygon, cone, and pyramid.

13. The method of claim 5, wherein the temperature control body further comprises a thermal barrier between the sectors.

14. The method of claim 5, wherein said chemical reaction is a polymerase chain reaction.

23. The method of claim 5, wherein said chemical reaction is a polymerase chain reaction.

15. A method for continuously regulating temperature of a fluid for production of a product, comprising:

dispensing a reaction mixture into a tubing having a first end, a second end, and a length, and said tubing further defining a volume, wherein the first end of said tubing extends from a first end of a channel and the second end of said tubing extends from a second end of said channel, said reaction mixture being dispensed into the second end of said tubing, wherein said channel spirals around a perimeter of a temperature control body comprising at least two sectors that each form a portion of the perimeter, wherein each sector has at least one temperature control means and is substantially made of a solid material so as to be configured to operate as an independent temperature sink;

conveying said reaction mixture through said tubing from the second end of said tubing to the first end of said tubing;

determining a temperature of said tubing as said reaction mixture flows through said tubing across each said sector of said temperature control body;

regulating said at least one temperature control means of each sector based on the determined temperature so as to achieve a target temperature for the sector;

continuously replenishing said reaction mixture into the second end of said tubing so that said reaction mixture is continuously conveyed through the length of said tubing, so that said reaction mixture fills the entire volume of said tubing along the length of said tubing from the second end to the first end, and so that said reaction mixture is conveyed without impediment from the second end to the first end; and

collecting a continual supply of said product at the first end of said tubing so long as the reaction mixture is being conveyed through said tubing.

16. The method of claim 15, wherein said temperature control body is a cylinder having a circumference, wherein each sector is wedge-shaped, and wherein said channel spirals around the circumference of the cylinder by one of (i) boring through said sectors internally from one sector to each successive sector, (ii) passing along the exterior surface of the cylinder from one sector to each successive sector, and (iii) alternating between boring through one or more successive sectors and passing along the exterior surface of the cylinder so as to traverse one or more successive sectors.

17. The method of claim 15, wherein said temperature control body is a cylinder having a circumference and a longitudinal axis, wherein the sectors are split into discontinuous layers, each sector being split along a plane perpendicular to the longitudinal axis so that successive sectors are layered adjacent to one another along the longitudinal axis of the cylinder.

18. The method of claim 15, wherein said first end of said channel terminates near a top edge of said temperature control body and said second end of said channel terminates near a bottom edge of said temperature control body.

19. The method of claim 15, wherein each sector is substantially made of a thermal conductor.

20. The method of claim 19, wherein said thermal conductor is selected from the group consisting of aluminum, aluminum alloy, metal, alloy, ceramic, and combinations thereof.

21. The method of claim 15, wherein said temperature control body is surrounded with at least one insulating layer.

22. A method for the facilitation of a chemical reaction requiring cyclical temperature changes for production of a product, comprising:

activating one or more means for changing temperature on a thermal cycle system, wherein said thermal cycle system comprises

a temperature control body comprising an exterior surface, at least two sectors forming a portion of said exterior surface, and a path cyclically passing through said sectors, wherein each said sector comprises at least one of said means for changing temperature and is an independent temperature sink,

a tubing for conveying a fluid having a length, a first end, and a second end, said tubing further defining a volume and extending along said path, and

a pump for moving said fluid through said tubing;

introducing a fluid, which is a substantially homogeneous temperature-dependent reaction mixture that includes a non-ionic surfactant, into said tubing;

activating said pump such that said reaction mixture moves through said tubing, and such that said reaction mixture reacts to form a product;

continuously replenishing said reaction mixture at the second end of said tubing while said pump is activated so that said reaction mixture is continuously conveyed through the length of said tubing, so that said reaction mixture fills the entire volume of said tubing along the length of said tubing from the second end to the first end, and so that said reaction mixture is conveyed without impediment from the second end to the first end; and

collecting a continual supply of said product at said first end of said tubing so long as said pump is activated.