Thermal cycler for automatic performance of the polymerase chain reaction with close temperature control

• US 7,504,241 B2
• Filed: 05/12/2006
• Issued: 03/17/2009
• Est. Priority Date: 11/29/1990
• Status: Expired due to Fees

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Inventors: Ragusa, Robert P., Woudenberg, Timothy M., Leath, Richard, Mossa, Albert Carmelo, Atwood, John Girdner, Goven, Lisa May, Williams, Fenton, Margulies, Marcel, Miles, Clive
• Primary Examiner(s): Beisner; William H
• Application Number:
  US11/433,872
• Publication Number:
  US 20060286659A1
• Time in Patent Office: 1,040 Days
• Field of Search: None
• US Class Current: 435/91.2
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/042 Caps; Plugs
• CPC Class Codes: B01L 2300/046 Function or devices integrated in the closure
• 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 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: B01L 7/00 Heating or cooling apparatus autoclaves B01J3/04; Heat insulating devices
• 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: G05D 23/1917 using digital means
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

A thermal cycler for automatic performance of the polymerase chain reaction is provided. The thermal cycler comprises a heater control that provides close temperature control of the reaction.

Claims

1. A method comprising:

positioning a sealed vessel in at least one sample well, the sealed vessel comprising a top and containing a polymerase chain reaction sample mixture having a concentration, wherein the polymerase chain reaction mixture comprises DNA and at least two primers complementary to the DNA to create extension products of the DNA;

thermal cycling the polymerase chain reaction sample mixture in the sealed vessel received in the sample well, through many polymerase chain reaction cycles; and

contacting the top of the sealed vessel with the platen while heating the platen so that the platen is kept at a temperature from 94°

C. to 110°

C. during the many polymerase chain reaction cycles and condensation and evaporation of the sample mixture within the sealed vessel is avoided during the many polymerase chain reaction cycles such that the polymerase chain reaction sample mixture concentration is controlled through the many polymerase chain reaction cycles.

2. The method of claim 1, wherein the at least one sample well comprises a plurality of sample wells, a plurally of sealed vessels are received in the plurality of sample wells, and the plurality of sealed vessels comprise a microtiter plate format.

3. The method of claim 1, wherein the platen is kept at a temperature of from 100°

C. to 110°

C.

4. The method of claim 1, wherein the many polymerase chain reaction cycles comprises 25 to 30 cycles.

5. The method of claim 1, wherein the at least one sample well comprises a plurality of sample wells, and the sealed vessel comprises a plurality of sealed vessels received in the plurality of sample wells.

6. The method of claim 5, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each of the sample tubes comprises a cap, and the contacting comprises forcing the bottom surface of the platen against the caps.

7. The method of claim 6, wherein the contacting comprises deforming the caps.

8. The method of claim 6, wherein the contacting comprises pressing each sample tube into its respective sample well.

9. The method of claim 5, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each sample well comprises an inside surface, and the contacting comprises snugly contacting a lower portion of each sample tube with the respective inside surface of the sample well that contains the lower portion.

10. A method comprising:

positioning a sealed vessel in at least one sample well, the sealed vessel comprising a top and containing a polymerase chain reaction sample mixture having a concentration, wherein the polymerase chain reaction mixture comprises DNA and at least two primers complementary to the DNA to create extension products of the DNA;

thermal cycling the polymerase chain reaction sample mixture in the sealed vessel received in the sample well, through many polymerase chain reaction cycles; and

contacting the top of the sealed vessel with the platen while heating the platen so that the platen is heated to a temperature above the boiling point of water during the many polymerase chain reaction cycles and condensation and evaporation of the sample mixture within the sealed vessel is avoided during the many polymerase chain reaction cycles such that the polymerase chain reaction sample mixture concentration is controlled through the many polymerase chain reaction cycles.

11. The method of claim 10, wherein the at least one sample well comprises a plurality of sample wells, a plurality of sealed vessels are received in the plurality of sample wells, and the plurality of sealed vessels comprise a microtiter plate format.

12. The method of claim 10, wherein the many polymerase chain reaction cycles comprises 25 to 30 cycles.

13. The method of claim 10, wherein the at least one sample well comprises a plurality of sample wells, and the sealed vessel comprises a plurality of sealed vessels received in the plurality of sample wells.

14. The method of claim 13, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each of the sample tubes comprises a cap, and the contacting comprises forcing the platen against the caps.

15. The method of claim 14, wherein the contacting comprises deforming the caps.

16. The method of claim 14, wherein the contacting comprises pressing each sample tube into its respective sample well.

17. The method of claim 13, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each sample well comprises an inside surface, and the contacting comprises snugly contacting a lower portion of each sample tube with the respective inside surface of the sample well that contains the lower portion.

18. A method comprising:

thermal cycling at least one sample well in which is received a sealed vessel, the sample well comprising a wall that terminates at a top surface, the sealed vessel comprising a tube and a cap and containing a polymerase chain reaction sample mixture having a concentration, the cap projecting above the top surface, wherein the polymerase chain reaction mixture comprises DNA and at least two primers complementary to the DNA to create extension products of the DNA;

contacting the cap of the sealed vessel with a bottom surface of a platen while heating the platen;

deforming the cap by forcing the bottom surface of the platen to a position on a reference plane above and spaced from the top surface such that the sealed vessel is pushed into the sample well; and

heating the platen so that the platen is kept at a temperature from 94°

C. to 110°

C. during many polymerase chain reaction cycles and condensation and evaporation of the sample mixture within the sealed vessel is avoided during the many polymerase chain reaction cycles such that the polymerase chain reaction sample mixture concentration is controlled through the many polymerase chain reaction cycles.

19. The method of claim 18, wherein the at least one sample well comprises a plurality of sample wells, a plurality of sealed vessels are received in the plurality of sample wells, and the plurality of sealed vessels comprise a microtiter plate format.

20. The method of claim 18, wherein the platen is kept at a temperature of from 100°

C. and 110°

C.

21. The method of claim 18, wherein the at least one sample well comprises a plurality of sample wells, and the sealed vessel comprises a plurality of sealed vessels received in the plurality of sample wells.

22. The method of claim 21, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each of the sample tubes comprises a cap, and the contacting comprises forcing the bottom surface of the platen against the caps.

23. The method of claim 22, wherein the contacting comprises deforming the caps.

24. The method of claim 22, wherein the contacting comprises pressing each sample tube into its respective sample well.

25. The method of claim 21, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each sample well comprises an inside surface, and the contacting comprises snugly contacting a lower portion of each sample tube with the respective inside surface of the sample well that contains the lower portion.

26. A method comprising:

thermal cycling at least one sample well in which is received a sealed vessel, the sample well comprising a wall that terminates at a top surface, the sealed vessel comprising a tube and a cap and containing a polymerase chain reaction sample mixture having a concentration, the cap projecting above the top surface, wherein the polymerase chain reaction mixture comprises DNA and at least two primers complementary to the DNA to create extension products of The DNA;

contacting the cap of the sealed vessel with a bottom surface of a platen while heating the platen;

deforming the cap by forcing the bottom surface of the platen to a position on a reference plane above and spaced from the top surface such that the sealed vessel is pushed into the sample well; and

heating the platen so that the platen is heated to a temperature above the boiling point of water during many polymerase chain reaction cycles and condensation and evaporation of the sample mixture within the sealed vessel is avoided during the many polymerase chain reaction cycles such that the polymerase chain reaction sample mixture concentration is controlled through the many polymerase chain reaction cycles.

27. The method of claim 26, wherein the at least one sample well comprises a plurality of sample wells, a plurality of sealed vessels are received in the plurality of sample wells, and the plurality of sealed vessels comprise a microtiter plate format.

28. The method of claim 26, wherein the at least one sample well comprises a plurality of sample wells, and the sealed vessel comprises a plurality of sealed vessels received in the plurality of sample wells.

29. The method of claim 28, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each of the sample tubes comprises a cap, and the contacting comprises forcing the platen against the caps.

30. The method of claim 29, wherein the contacting comprises deforming the caps.

31. The method of claim 29, wherein the contacting comprises pressing each sample tube into its respective sample well.

32. The method of claim 28, wherein the plurality of sealed vessels comprises a plurality of sample tubes, each sample well comprises an inside surface, and the contacting comprises snugly contacting a lower portion of each sample tube with the respective inside surface of the sample well that contains the lower portion.