Clamp for fast PCR heating

• US 10,315,198 B2
• Filed: 01/17/2017
• Issued: 06/11/2019
• Est. Priority Date: 05/24/2012
• Status: Active Grant

• Current Assignee: BJS IP Ltd.
• Original Assignee: BJS IP Ltd.
• Inventors: Burroughs, Nicholas, Gunter, Ian, Lewis, Richard
• Primary Examiner(s): Hobbs, Michael L
• Application Number:
  US15/408,313
• Publication Number:
  US 20170189911A1
• Time in Patent Office: 875 Days
• Field of Search: 4353031
• CPC Class Codes: B01L 2200/02 Adapting objects or devices to another
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• 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 9/50 Clamping means, tongs in general F16B2/06
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]

Abstract

This disclosure provides a thermocycler system for performing polymerase chain reaction. The thermocycler system can comprise a plurality of bus bars, a microplate, a clamp and a current application device in electrical communication with the bus bars. The clamp is suitable for forming a low resistance electrical connection between the microplate and an individual bus bar of the plurality of bus bars. In some instances, the clamp includes a spring-loaded pivot and a ram. Operation of the clamp forces the microplate into contact with the bus bar, or vice versa, to provide an electrical contact between the microplate and the bus bar. The microplate can comprise a substrate having a metallic material for heating samples, and a barrier layer disposed adjacent to the substrate.

Claims

1. A method for forming a low resistance electrical connection between a microplate and a busbar in a thermocycler, comprising:

(a) providing a clamp comprising a spring-loaded pivot coupled to a ram; and

(b) using the spring-loaded pivot to provide linear motion to the ram to press the microplate against the bus-bar.

2. The method of claim 1, wherein the clamp is an over center toggle type clamp.

3. The method of claim 1, wherein the clamp provides control over a force applied by the ram over a range of adjustments.

4. The method of claim 1, wherein the clamp is closed by hand to form the low resistance electrical connection between the microplate and the bus-bar.

5. The method of claim 1, wherein the clamp is capable of exerting a force of between about 1,500 and 2,000 Newtons to the microplate and/or the bus-bar.

6. The method of claim 1, wherein the clamp deforms the microplate, thereby disrupting an oxide film on a surface of the microplate.

7. The method of claim 1, wherein the low resistance electrical resistance at an interface between the microplate and the bus-bar is less than a resistance of the microplate.

8. The method of claim 1, wherein the microplate is consumable.

9. The method of claim 1, wherein the microplate comprises:

(a) a substrate comprising a metallic material for heating a sample during polymerase chain reaction (PCR);

(b) a barrier layer disposed adjacent to the substrate, the barrier layer formed of a first polymeric material; and

(c) one or more wells for holding the sample during PCR, the one or more wells formed of a second polymeric material sealed to the barrier layer,

wherein the substrate provides a PCR ramp rate of at least 5°

C./second.

10. The method of claim 9, wherein the microplate heats the sample upon flow of electric current through the substrate.

11. The method of claim 9, wherein the metallic material has a resistivity between about 2×

10<sup>−

8</sup> ohm-m and 8×

10<sup>−

8</sup> ohm-m.

12. The method of claim 9, wherein the metallic material comprises aluminum.

13. The method of claim 9, wherein the microplate has a thickness of less than 1 mm.

14. The method of claim 9, wherein the substrate increases a temperature of the sample at a rate between about 5°

C./s and about 15°

C./s.

15. The method of claim 9, wherein the one or more wells comprise at least 24 wells.

16. The method of claim 9, wherein the one or more wells comprise at least 96 wells.

17. The method of claim 9, wherein the substrate provides well-to-well thermal uniformity of at least about +/−

0.5°

C. without an external heating element or a Peltier heating block.

18. The method of claim 9, wherein the substrate provides a heating efficiency sufficient for at least about 2 PCR cycles per minute, including fluorescence measurements for every cycle.

19. The method of claim 9, wherein the barrier layer has a thickness of less than 10 micrometers.

20. The method of claim 1, further comprising (c) providing a sample in the microplate and (d) using the microplate to heat the sample at a rate of at least about 10°

C./second.