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Microfluidic devices and systems for performing inefficient fast PCR    
United States Patent6524830   
Link to this pagehttp://www.wikipatents.com/6524830.html
Inventor(s)Kopf-Sill; Anne R. (Portola Valley, CA)
AbstractMethods 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.



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Patent Text Patent PDF Print Page Summary File History
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Inventor     Kopf-Sill; Anne R. (Portola Valley, CA)
Owner/Assignee     Caliper Technologies Corp. (Mountain View, CA)
Patent assignment
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Publication Date     February 25, 2003
Application Number     09/943,070
PAIR File History     Application Data   Transaction History
Image File Wrapper   Patent Term   Fees
Litigation
Filing Date     August 29, 2001
US Classification     435/91.2 435/287.2 435/6 435/7.1 435/91.1 536/22.1 536/23.1 536/24.3 536/24.31 536/24.32 536/24.33
Int'l Classification    
Examiner     Siew; Jeffrey
Assistant Examiner    
Attorney/Law Firm     Filler; Andrew L.
Address
Parent Case     CROSS-REFERENCES TO RELATED APPLICATIONS This application is a divisional of application U.S. Ser. No. 09/287,069 filed Apr. 6, 1999, now U.S. Pat. No. 6,303,343. The present application claims priority to and benefit of this prior application.
Priority Data    
USPTO Field of Search     435/6 435/7.1 435/91.1 435/91.2 435/287.2 536/22.1 536/23.1 536/24.3 536/4 536/5 536/6 536/7 536/8 536/9 536/10 536/11 536/12 536/13 536/14 536/15 536/16 536/17 536/18 536/19 536/20 536/21 536/22 536/23 536/24 536/25 536/26 536/27 536/28 536/29 536/30 536/31 536/32 536/33
Patent Tags     microfluidic devices performing inefficient fast pcr
   
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6180372
Franzen

Jan,2001
[0 after 0 votes]		6140086
Fox et al.

Oct,2000
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Oct,1999
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Mar,1999
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Parce et al.

Feb,1999
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Jan,1999
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5800690
Chow et al.

Sep,1998
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What is claimed is:

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 .mu.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.

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 .mu.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.

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.

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.
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FIELD OF THE INVENTION

This invention is in the field of cyclic polymerase-mediated reactions such as PCR. More specifically, this invention relates to altering the time within which the steps of such reactions are carried out. The methods of this invention are particularly relevant to reactions such as PCR as performed in devices that allow very short cycle times, such as microfluidic devices.

BACKGROUND OF THE INVENTION

PCR and other cyclic polymerase-mediated reactions are standard tools of modern biological research, and are also commonly used for numerous applications including medical diagnostic procedures and forensic applications. PCR is based on three discrete, multiply repeated steps: denaturation of a DNA template, annealing of a primer to the denatured DNA template, and extension of the primer with a polymerase to create a nucleic acid complementary to the template. The conditions under which these steps are performed are well established in the art.

Generally, standard PCR protocols teach the use of a small number of cycles (e.g. 20-35 cycles) which are optimized for maximum efficiency in each cycle, i.e. to ensure that a highest possible percentage of template molecules is copied in each cycle. Typically, this entails cycle times of 1.2, or more minutes. For example, the standard reference Innis et al., PCR Potocols, A Guide to Methods and Applications (Academic Press, Inc.; 1990)("Innis") suggests the following conditions under the heading "Standard PCR Amplification Protocol" (at page 4):

Perform 25 to 35 cycles using the following temperature profile:

Denaturation 96.degree. C., 15 seconds Primer Annealing 55.degree. C., 30 seconds Primer Extension 72.degree. C., 1.5 minutes

Such times, or longer, are typical in the field. Similar protocols can be found in, e.g. Sambrook et al. (1989) Molecular Cloning, A Laboratory Manual (2d Edition), Cold Spring Harbor Press, Cold Spring Harbor, N.Y. ("Sambrook"), which teaches a 6 minute cycle, and Ausubel et al., eds. (1996) Cur