Thermal cycler including a temperature gradient block

• US 20020127660A1
• Filed: 03/02/2001
• Published: 09/12/2002
• Est. Priority Date: 10/20/1993
• Status: Abandoned Application

• Current Assignee: Stratagene California
• Original Assignee: Stratagene California
• Inventors: Shopes, Robert James, Danssaert, John Lewis, Shoemaker, Daniel Davis
• Application Number:
  US09/796,599
• Publication Number:
  US 20020127660A1
• Time in Patent Office: Days
• US Class Current: 435/91.3
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1838 using fluid heat transfer medium
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: B01L 7/5255 by moving sample containers
• CPC Class Codes: B01L 7/54 using spatial temperature gradients
• CPC Class Codes: G01N 2035/042 moved independently, e.g. by fork manipulator
• CPC Class Codes: G01N 35/0099 comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

A method in which a temperature gradient is generated across a “gradient” block, and an apparatus comprising a block across which a temperature gradient can be generated. By setting up such a gradient, multiple reaction mixtures held in wells on the gradient block can be simultaneously run at temperatures which differ only slightly, thereby permitting an optimum temperature for the reaction to be quickly identified. In a preferred embodiment the gradient block is integrated into a thermal cycler used for nucleic acid amplification reactions.

Claims

1. A method for simultaneously reacting a plurality of reaction mixtures in an apparatus including a temperature gradient block comprising the steps of:

placing reaction mixtures in a plurality of reaction wells in said gradient block, said gradient block having a top portion, first and second oppposing portions, and a bottom portion, said plurality of reaction mixture wells being formed in said block between said opposing portions, and

generating a temperature gradient across said gradient block and between said opposing portions.

2. A method according to claim 1 Wherein said step of generating a temperature gradient comprises the steps of heating said first opposing portion of said gradient block, and cooling said second opposing portion of said gradient block.

3. A method according to claim 1 including the further step of controlling said temperature gradient using controlling means.

4. A method according to claim 3,wherein said controlling step comprises the steps of collecting and storing temperature set point and actual temperature data from said wells, and transmitting said information to a microprocessor.

5. A method according to claim 1 wherein said apparatus further comprises at least one additional heat conducting block having a top portion, first and second opposing portions, and a bottom portion, and a plurality of reaction mixture wells formed in said additional block between said opposing portions, the method further comprising the step of moving said reaction mixtures between said gradient block and said additional block or blocks.

6. A method for automated temperature cycling of a reaction mixture using a thermal cycling apparatus comprising at least one heat conducting block, said block having a plurality of sample wells spaced between first and second opposing portions and in an upper surface thereof, the method comprising

placing reaction mixtures in said wells, and

generating a temperature gradient across said block and between said opposing portions by heating said first opposing portion and cooling said second opposing portion.

7. An apparatus for generating a temperature gradient across a heat conducting block comprising:

a reaction mixture holder, said reaction mixture holder comprising a heat conducting block having a top portion, first and second opposing portions, and a bottom portion, a plurality of reaction mixture wells formed in said top portion, and between said first and second opposing portions,

a block heater positioned adjacent to said first opposing portion, and

a block cooler positioned adjacent to said second opposing portion.

8. An apparatus for generating a temperature gradient across a heat conducting block according to claim 7 wherein said apparatus further comprises controller means for controlling said block heater and block cooler.

9. An apparatus for generating a temperature gradient across a heat conducting block according to claim 7 wherein said heat conducting block comprises brass.

10. An apparatus for generating a temperature gradient across a heat conducting block according to claim 8 wherein said controller means comprises a microprocessor for collecting and storing temperature set point and actual temperature data.

11. An apparatus for generating a temperature gradient across a heat conducting block according to claim 7 wherein said plurality of wells in said heat conducting block are spaced across said top portion.

12. An apparatus for generating a temperature gradient across a heat conducting block according to claim 11 wherein said plurality of wells in said heat conducting block are spaced across said top portion in parallel, aligned rows.

13. An apparatus for generating a temperature gradient across a heat conducting block according to claim 7 wherein said holder further comprises a heater positioned adjacent to said bottom portion.

15. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said means for generating a temperature gradient comprises means for heating said first opposing portion of said block, and means for cooling said second opposing portion of said block.

16. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said apparatus further comprises at least one additional holding means for holding a reaction mixture, said additional holding means comprising:

(i) at least one additional heat conducting block including a plurality of reaction mixture wells; and

(ii) means for heating said additional heat conducting block.

17. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said heat conducting block comprises brass.

18. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said plurality of wells in said heat conducting block are spaced across said block in parallel, aligned rows.

19. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said apparatus further comprises controller means for generating said temperature gradient.

20. An apparatus for generating a temperature gradient across a heat conducting block according to claim 19 wherein said controller means comprises a microprocessor for collecting and storing temperature is set point and actual temperature data.

21. An apparatus for generating a temperature gradient across a heat conducting block according to claim 14 wherein said holder means further comprises a heating means for heating said bottom portion.

22. An apparatus for generating a temperature gradient across a heat conducting block according to claim 16 wherein said apparatus further comprises robot arm means, controlled by robot arm control means, for moving said reaction mixture between said holding means.

14. An apparatus for generating a temperature gradient across a heat conducting block comprising holding means for holding a reaction mixture, said holding means comprising:

(i) a heat conducting block having a top portion, first and second opposing portions, and a bottom portion, and a plurality of reaction mixture wells formed in said top portion and between said first and second opposing portions; and

(ii) means for generating a temperature gradient across said heat conducting block and between said first and second opposing portions.

23. An automated apparatus for performing molecular biological reactions comprising at least one temperature controlled block, said block having a plurality of reaction mixture wells spaced between first and second opposing portions and in an upper portion thereof, and a block heater positioned adjacent to said first opposing portion and capable of generating a temperature gradient between said first and second opposing portions.