Microfluidic devices and systems for performing inefficient fast PCR

• US 6,524,830 B2
• Filed: 08/29/2001
• Issued: 02/25/2003
• Est. Priority Date: 04/06/1999
• Status: Expired due to Term

• Current Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Original Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Inventors: Kopf-Sill, Anne R.
• Primary Examiner(s): Siew, Jeffrey
• Application Number:
  US09/943,070
• Publication Number:
  US 20020055149A1
• Time in Patent Office: 545 Days
• Field of Search: 435/6, 435/7.1, 435/91.1, 435/91.2, 435/287.2, 536/22.1, 536/23.1, 536/24-- 3
• US Class Current: 435/91.2
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• 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 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2565/629 being a microfluidic device

Abstract

Methods of performing fast polymerase mediated reactions are provided. These reactions can be used in an inefficient fashion in the cycles of the polymerase mediated reactions to produce product at a much faster rate than conventional polymerase mediated reaction methods. Integrated systems for performing these methods are also provided.

Claims

1. An apparatus for amplifying one or more nucleic acids, comprising,

a body with at least one microscale channel fabricated therein;

and,

a thermal control element coupled to the microscale channel, which control element cycles the temperature at least 50 times in one or more regions of the microscale channel at cycle times ranging between 5 and 20 seconds.

2. The apparatus of claim 1, wherein the at least one microscale channel comprises at least sixty temperature-controlled zones.

3. The apparatus of claim 1, wherein the diameter of at least one of said at least one channels is between 0.1 and 500 μ

m.

4. The apparatus of claim 1, wherein said temperature is controlled by modulating the current per cross sectional dimension in a region of the channel.

5. The apparatus of claim 1, wherein said temperature is controlled by modulating a current in at least one fluid region of the microscale channel which are at different temperatures.

6. The apparatus of claim 1, comprising a source of test samples fluidly connected to said at least one channel.

7. The apparatus of claim 1, comprising a detection zone fluidly connected to said at least one channel.

8. The apparatus of claim 1, further comprising at least one receptacle for collecting at least one of said amplified nucleic acids, which at least one receptacle is fluidly connected to said microscale channel.

21. The apparatus of claim 1, wherein the control element cycles the temperature at least 70 times.

22. The apparatus of claim 1, wherein the control element cycles the temperature at least 100 times.

23. The apparatus of claim 1, wherein the control element cycles the temperature at least 200 times.

24. The apparatus of claim 1, wherein the control element cycles the temperature at least 400 times.

25. The apparatus of claim 1, wherein the control element cycles the temperature at least 1,000 times.

9. An integrated system for amplifying one or more nucleic acids, comprising:

an apparatus comprising a body with at least one microscale channel fabricated therein and a thermal control element coupled to the microscale channel;

a computer and software for controlling one or more of;

(i) temperature of at least one fluid within said microscale channel, which thermal control element cycles the temperature in one or more regions of the microscale channel for at least 50 cycles at cycle times ranging between 5 and 20 seconds;

(ii) number of different temperature-controlled zones within said microscale channel; and

(iii) movement of at least one fluid present within said microscale channel.

10. The integrated system of claim 9, wherein the apparatus comprises at least sixty temperature-controlled zones.

11. The integrated system of claim 9, wherein the diameter of at least one of said at least one microscale channel is between 0.1 and 500 μ

m.

12. The integrated system of claim 9, wherein the temperature of the at least one fluid within the apparatus is controlled by modulating the current per cross sectional dimension in a region of the channel.

13. The integrated system of claim 9, wherein the temperature of the at least one fluid within the apparatus is controlled by flowing the at least one fluid into regions of the microscale channel which are at different temperatures.

14. The integrated system of claim 9, further comprising a source of test samples fluidly connected to the at least one microscale channel.

15. The integrated system of claim 9, further comprising a plurality of sources of test samples fluidly connected to said at least one microscale channel.

16. The integrated system of claim 15, wherein the computer controls the selection or introduction of the test samples into the at least one channel.

17. The integrated system of claim 9, further comprising a detection zone fluidly connected to the channel.

18. The integrated system of claim 17, wherein the computer controls the detection or analysis of data from the detection zone.

19. The integrated system of claim 9, further comprising at least one receptacle for collecting at least one of the amplified nucleic acids, which at least one receptacle is fluidly connected to the microscale channel.

20. The integrated system of claim 19, wherein the computer controls the collection of at least one of the amplified nucleic acids using said at least one receptacle.

26. The integrated system of claim 9, wherein the control cycles the temperature at least 70 times.

27. The integrated system of claim 9, wherein the control element cycles the temperature at least 100 times.

28. The integrated system of claim 9, wherein the control element cycles the temperature at least 200 times.

29. The integrated system of claim 9, wherein the control element cycles the temperature at least 400 times.

30. The integrated system of claim 9, wherein the control element cycles the temperature at least 1,000 times.