Thermocycling apparatus and method

• US 5,985,651 A
• Filed: 06/16/1997
• Issued: 11/16/1999
• Est. Priority Date: 06/17/1996
• Status: Expired due to Fees

• Current Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Original Assignee: Board of Trustees of the Leland Stanford Junior University (Stanford Management Co.)
• Inventors: Hunicke-Smith, Scott P.
• Primary Examiner(s): Redding, David A.
• Application Number:
  US08/877,361
• Time in Patent Office: 883 Days
• Field of Search: 435/283.1, 435/286.5, 435/287.2, 435/289.1, 435/285.1, 219/535, 219/201, 219/528, 392/479, 392/480, 392/482, 165/185, 165/177, 165/182, 165/183, 422/81
• US Class Current: 435/285.1
• CPC Class Codes: B01L 3/0217 of the plunger pump type medical syringes A61M
• 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 7/5255 by moving sample containers
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks

Abstract

A thermocycling apparatus comprising a plurality of capillaries for moving DNA-containing samples between two or more discrete zones maintained at selected elevated temperatures.

Claims

1. An apparatus for thermally cycling a DNA sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) means for maintaining the temperatures of the first and second chambers of said heating unit assembly at selected first and second elevated temperatures; and

(iv) means for moving the sample in the tube successively between the two chambers;

wherein the sample moving means is effective to move the sample in a reciprocating motion within the tubes.

2. The apparatus of claim 1, wherein one elevated temperature is effective to denature the DNA sample and the other elevated temperature is one at which DNA annealing and primer-directed DNA polymerization can occur.

3. The apparatus of claim 1, wherein said moving means includes a syringe body, a plunger, and means for moving the plunger in a reciprocating motion within the syringe body.

4. The apparatus of claim 3, wherein the moving means includes an integrated circuit (IC) motion controller and a movement unit.

5. The apparatus of claim 4, wherein the moving means further includes a sample position sensor.

6. The apparatus of claim 5, wherein the sample position sensor is an optical sensor.

7. The apparatus of claim 4, wherein said movement unit includes an IC motor driver, a direct current (DC) motor with a threaded rod, and a syringe body with a plunger.

8. The apparatus of claim 1, wherein the temperature maintenance means includes a temperature controller, a power delivery system, and a temperature feedback system.

9. The apparatus of claim 8, wherein the temperature controller is an integrated circuit (IC) temperature controller, and the power delivery system includes an IC heater driver a power transistor.

10. The apparatus of claim 8, wherein the temperature feedback system includes an analog-to-digital (A/D) convertor and a separate thermistor temperature sensor for each heating element.

11. The apparatus of claim 1, for thermally cycling a plurality of DNA samples, comprising a plurality of heating unit assemblies mounted on said support.

12. The apparatus of claim 11, wherein the sample moving means is effective to move the samples in all capillaries in a concerted manner.

13. The apparatus of claim 11, wherein the moving means is effective to move the samples independently of one another.

14. The apparatus of claim 11, wherein temperatures of the heating chambers of each heating unit assembly are maintained independently of temperatures of the heating chambers of other heating unit assemblies.

15. The apparatus of claim 11, wherein the apparatus contains 8 linearly-disposed heating unit assemblies spaced at approximately 9 mm intervals.

16. The apparatus of claim 11, wherein the apparatus contains 12 linearly-disposed heating unit assemblies spaced at approximately 9 mm intervals.

17. The apparatus of claim 11, wherein the heating unit assemblies, sample moving means and temperature maintenance means are all mounted on said support.

18. The apparatus of claim 17, wherein said support is a module board and said apparatus is adapted for use in an instrument capable of receiving a plurality of such module boards.

19. The apparatus of claim 1, wherein

(i) the heating unit assembly further contains a third heating element defining a third heating chamber,

(ii) the temperature maintenance means is effective to maintain the temperature of said third heating chamber at a selected third elevated temperature, and

(iii) the sample moving means includes means for moving the sample in the tube successively between the three chambers of the heating unit assembly.

20. The apparatus of claim 19, wherein the first elevated temperature is effective to denature the DNA sample, the second elevated temperature is one at which DNA annealing can occur, and the third elevated temperature is one at which primer-directed DNA polymerization can occur.

21. The apparatus of claim 1, wherein

(i) the heating unit assembly further contains a cooling element defining a cooling chamber,

(ii) the temperature maintenance means is effective to maintain the temperature of said cooling chamber at a selected depressed temperature, and

(iii) the sample moving means includes means for moving the sample in the tube successively between the heating and cooling chambers of the heating unit assembly.

22. The apparatus of claim 21, wherein the depressed temperature is between about 0°

C. and about 10°

C.

23. An apparatus for thermally cycling DNA samples, comprising

(i) a plurality of capillary assemblies, each assembly comprising first and second heating elements, defining first and second heating chambers, respectively,

a capillary tube extending through the two heating chambers, and

means for introducing one such DNA sample into the capillary tube;

(ii) means for maintaining the temperatures of the first and second chambers of such a capillary assembly at selected first and second elevated temperatures, wherein one elevated temperature is effective to denature the DNA sample and the other elevated temperature is one at which DNA annealing and primer-directed DNA polymerization can occur; and

(iii) means for moving the samples in the tubes successively between the two chambers of each capillary assembly; and

wherein the capillary tubes have an interior face that is hydrophobic.

24. The apparatus of claim 23, wherein the sample volume in each capillary is between about 0.5 and 5 μ

l.

25. An apparatus for thermally cycling a DNA sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) means for maintaining the temperatures of the first and second chambers of said heating unit assembly at selected first and second elevated temperatures; and

(iv) means for moving the sample in the tube successively between the two chambers;

wherein said moving means includes a syringe body, a plunger, and means for moving the plunger in a reciprocating motion within the syringe body.

26. An apparatus for thermally cycling a DNA sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) means for maintaining the temperatures of the first and second chambers of said heating unit assembly at selected first and second elevated temperatures; and

(iv) means for moving the sample in the tube successively between the two chambers;

wherein the sample moving means is effective to move the samples in all capillaries in a concerted manner.

27. An apparatus for thermally cycling a DNA sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) means for maintaining the temperatures of the first and second chambers of said heating unit assembly at selected first and second elevated temperatures; and

(iv) means for moving the sample in the tube successively between the two chambers; and

wherein the heating unit assemblies, sample moving means and temperature maintenance means are all mounted on said support.

28. An apparatus for thermally cycling a nucleic acid sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) a temperature control in communication with said heating elements; and

(iv) a sample position control unit disposed in communication with said tube and operable to alter pressure along one end thereof, said altering of pressure being effective to move a nucleic acid sample in the tube successively and reciprocally between the two chambers.

29. An apparatus for thermally cycling a nucleic acid sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

wherein said capillary tube has an interior face that is hydrophobic;

(iii) a temperature control in communication with said heating elements; and

(iv) a sample position control unit disposed in communication with said tube and operable to alter pressure along one end thereof, said altering of pressure being effective to move a nucleic acid sample in the tube successively between the two chambers.

30. An apparatus for thermally cycling a nucleic acid sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) a temperature control in communication with said heating elements; and

(iv) a sample position control unit, including a syringe body and a plunger movable within the syringe body, disposed in communication with said tube;

wherein movement of said plunger is effective to alter pressure along one end of said capillary tube, thereby moving a nucleic acid sample in the tube successively between the two chambers.

31. An apparatus for thermally cycling nucleic acid samples, comprising

(i) a support;

(ii) a plurality of heating unit assemblies mounted on said support, each such assembly having first and second heating elements defining first and second heating chambers, respectively, said first and second heating chambers being adapted to receive and contact a respective capillary tube extending through the two heating chambers;

(iii) a temperature control in communication with said heating elements; and

(iv) a sample position control unit in communication with each capillary tube and operable to alter pressure along one end thereof, said altering of pressure being effective to move all the nucleic acid samples in the tubes successively between the two chambers in a concerted manner.

32. An apparatus for thermally cycling a nucleic acid sample, comprising

(i) a support;

(ii) a heating unit assembly mounted on said support, said assembly having first and second heating elements defining first and second heating chambers, respectively, said chambers being adapted to receive and contact a capillary tube extending through the two heating chambers;

(iii) a temperature control mounted on said support and disposed in communication with said heating elements; and

(iv) a sample position control unit mounted on said support in communication with said tube and operable to alter pressure along one end of said tube, said altering of pressure being effective to move a nucleic acid sample in the tube successively between the two chambers.