SYSTEM AND METHOD FOR TEMPERATURE REFERENCING FOR MELT CURVE DATA COLLECTION

• US 20090318306A1
• Filed: 06/23/2008
• Published: 12/24/2009
• Est. Priority Date: 06/23/2008
• Status: Active Grant

• Current Assignee: Canon USA, Inc. (Canon Inc.)
• Original Assignee: Canon US Life Sciences Inc (Canon Inc.)
• Inventors: Inoue, Hiroshi, Hasson, Kenton C.
• Granted Patent: US 9,393,566 B2
• Time in Patent Office: Days
• US Class Current: 506/12
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/502784 specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• 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: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2527/107 Temperature of melting, i.e. Tm
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/645 Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 2201/06113 Coherent sources; lasers
• CPC Class Codes: G01N 25/4866 by using a differential method
• CPC Class Codes: G01N 25/4893 by using a differential method

Abstract

The present invention relates to systems and methods of temperature referencing for melt curve data collection. More specifically, the present invention relates to systems and methods for collecting DNA melt curve data for a DNA sample and a temperature reference material.

Claims

1. A system for performing melt curve data collection comprising:

a container, a heating system and a detection system, wherein the container comprises at least two chambers that are in close thermal connection and wherein at least one chamber contains a DNA sample to be tested and at least one chamber contains a temperature reference material.

2. The system of claim 1, wherein the heating system supplies heat to all of the chambers simultaneously.

3. The system of claim 1, wherein the chambers contain at least two spatially separated temperature reference materials to determine a spatial temperature gradient.

4. The system of claim 1, wherein the temperature reference materials bracket the DNA sample in both space and time.

5. The system of claim 1, wherein the temperature reference material is mixed with the DNA sample.

6. The system of claim 1, wherein the detection system is an optical detection system.

7. The system of claim 1, wherein a detectable signal from the temperature reference material provides feedback to the heating system.

8. The system of claim 1, wherein the container is a microfluidic chip and the chambers are microfluidic channels in the chip.

9. The system of claim 1, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated from each other and contain a temperature reference material to determine a spatial temperature gradient.

10. The system of claim 9, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

11. The system of claim 1, wherein the temperature reference material is a known DNA mixture.

12. The system of claim 1, wherein the temperature reference material is a thermochromatic material.

13. A system for performing melt curve data collection, said system comprising:

a container, a heating system and a detection system, wherein the container comprises at least one chamber that is refillable without moving the container, and wherein the container contains a DNA sample to be tested and a temperature reference material.

14. The system of claim 13, wherein the heating system supplies heat to all of the chambers simultaneously.

15. The system of claim 13, wherein the detection system is an optical detection system.

16. The system of claim 13, wherein a detectable signal from the temperature reference material provides feedback to the heating system.

17. The system of claim 13, wherein the DNA sample and the temperature reference material are alternated in the chamber.

18. The system of claim 13, wherein the temperature reference materials bracket the DNA sample in both space and time.

19. The system of claim 13, wherein the DNA sample and the temperature reference material are mixed, wherein the temperature reference material has a detection signature that is discernible from that of the DNA sample, and wherein data from both the sample and the temperature reference material are collected at the same place and the same time.

20. The system of claim 13, wherein the temperature reference material is also a flow marker.

21. The system of claim 13, wherein the container is a microfluidic chip and the chambers are microfluidic channels in the chip.

22. The system of claim 21, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated and contain a temperature reference material to determine a spatial temperature gradient.

23. The system of claim 22, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

24. The system of claim 13, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated and contain a temperature reference material to determine a spatial temperature gradient.

25. The system of claim 24, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

26. The system of claim 13, wherein the temperature reference material is a known DNA mixture.

27. The system of claim 13, wherein the temperature reference material is a thermochromatic material.

28. A method of performing melt curve data collection comprising:

(a) providing a container which comprises at least two chambers that are in close thermal connection;

(b) introducing a DNA sample to be tested into at least one of the chambers and a temperature reference material into at least one of the other chambers;

(c) heating the chambers from a first temperature to a second temperature; and

(d) measuring a detectable property emanating from the DNA sample and a detectable property emanating from the temperature reference material, wherein the detectable property of the DNA sample indicates an extent of denaturation of the DNA in the sample and the detectable property of the temperature reference material is correlated to the temperature.

29. The method of claim 28, wherein the chambers contain at least two spatially separated temperature reference materials and the detectable property emanating from the temperature reference materials is measured to determine a spatial temperature gradient.

30. The method of claim 28, wherein the temperature reference materials bracket the DNA sample in both space and time.

31. The method of claim 30, wherein the detectable property emanating from the temperature reference materials is measured to determine a spatial temperature gradient and any temporal fluctuation.

32. The method of claim 28, wherein the temperature reference material is mixed with the DNA sample.

33. The method of claim 28, wherein the detectable property is measured with an optical detection system.

34. The method of claim 28, wherein the detectable property from the temperature reference material provides feedback to the heating system.

35. The method of claim 28, wherein the container is a microfluidic chip and the chambers are microfluidic channels in the chip.

36. The method of claim 28, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated from each other and contain temperature reference materials and the detectable property emanating from the temperature reference materials is measured to determine a spatial temperature gradient.

37. The method of claim 36, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

38. The method of claim 28, wherein the temperature reference material is a known DNA mixture.

39. The method of claim 28, wherein the temperature reference material is a thermochromatic material.

40. A method of performing melt curve data collection comprising:

(a) providing a container which comprises at least one chamber that is refillable without moving the container, wherein if the container comprises two or more chambers, the chambers are in close thermal connection;

(b) introducing a DNA sample to be tested into at least one of the chambers and a temperature reference material into at least one of the chambers;

(c) heating the chambers from a first temperature to a second temperature; and

(d) measuring a detectable property emanating from the DNA sample and a detectable property emanating from the temperature reference material, wherein the detectable property of the DNA sample indicates an extent of denaturation of the DNA and the detectable property of the temperature reference material is correlated to the temperature.

41. The method of claim 40, wherein the detectable property is measured with an optical detection system.

42. The method of claim 40, wherein a detectable property from the temperature reference material provides feedback to the heating system.

43. The method of claim 40, wherein the DNA sample and the temperature reference material are alternated in the same chamber.

44. The method of claim 40, wherein the temperature reference materials bracket the DNA sample in both space and time.

45. The method of claim 44 wherein the detectable property emanating from the temperature reference materials is measured to determine a spatial temperature gradient and any temporal fluctuation.

46. The method of claim 40, wherein the DNA sample and the temperature reference material are mixed, wherein the temperature reference material has a detection signature that is discernible from that of the DNA sample and wherein data from both the sample and the temperature reference material are collected at the same place and the same time.

47. The method of claim 40, wherein the temperature reference material is also a flow marker.

48. The method of claim 40, wherein the container is a microfluidic chip and the chambers are microfluidic channels in the chip.

49. The method of claim 48, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated and contain temperature reference materials and the detectable property emanating from the temperature reference materials is measured to determine a spatial temperature gradient.

50. The method of claim 49, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

51. The method of claim 40, wherein the container comprises at least three chambers wherein at least two of the chambers are spatially separated and contain a temperature reference material to determine a spatial temperature gradient.

52. The method of claim 51, wherein at least two chambers containing DNA samples are located between two chambers containing the temperature reference material.

53. The method of claim 40, wherein the temperature reference material is a known DNA mixture.

54. The method of claim 40, wherein the temperature reference material is a thermochromatic material.