MESOSCALE POLYNUCLEOTIDE AMPLIFICATION DEVICE AND METHOD

• US 20070190641A1
• Filed: 04/05/2007
• Published: 08/16/2007
• Est. Priority Date: 05/01/1992
• Status: Abandoned Application

• Current Assignee: Larry Kricka, Peter Wilding
• Original Assignee: Larry Kricka, Peter Wilding
• Inventors: Wilding, Peter, Kricka, Larry
• Application Number:
  US11/696,926
• Publication Number:
  US 20070190641A1
• Time in Patent Office: Days
• US Class Current: 435/287.1
• CPC Class Codes: B01D 2325/028 Microfluidic pore structures
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 67/003 by selective elimination of components, e.g. by leaching
• CPC Class Codes: B01D 67/0031 by elimination of at least one of the blocks of a block copolymer
• CPC Class Codes: B01D 71/02 Inorganic material
• CPC Class Codes: B01D 71/0213 Silicon
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/146 Employing pressure sensors
• 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 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• CPC Class Codes: B01L 2300/185 using a liquid as fluid
• CPC Class Codes: B01L 2300/1861 : using radiation
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502707 : characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction. The devices comprise a substrate microfabricated to define a sample inlet port and a mesoscale flow system, which extends from the inlet port. The mesoscale flow system includes a polynucleotide polymerization reaction chamber in fluid communication with the inlet port which is provided with reagents required for polymerization and amplification of a preselected polynucleotide. In one embodiment the devices may be utilized to implement a polymerase chain reaction (PCR) in the reaction chamber (PCR chamber). The PCR chamber is provided with the sample polynucleotide, polymerase, nucleoside triphosphates, primers and other reagents required for the polymerase chain reaction, and the device is provided with means for thermally controlling the temperature of the contents of the reaction chamber at a temperature controlled to dehybridize double stranded polynucleotide, to anneal the primers, and to polymerize and amplify the polynucleotide.

Claims

1. an analytical system for PCR analysis comprising:

a device comprising a mesoscale flow system and an inlet port communicating with the flow system;

an appliance comprising a nesting site that releasably holds the device, a flow line mating with the inlet port of the device, a pump connected to the flow line that when actuated forces fluid through the flow system of the device, and a sensor that detects properties of fluid within the flow system; and

means for thermally cycling fluid within the flow system.

2. The system of claim 1, wherein the device comprises a solid substrate.

3. The system of claim 2, wherein the solid substrate is approximately 0.2 to 2.0 centimeters square.

4. The system of claim 2, wherein the device further comprises a cover.

5. The system of claim 4, wherein the cover is a transparent cover.

6. The system of claim 2, wherein the solid substrate comprises silicon.

7. The system of claim 2, wherein the solid substrate comprises a polymer.

8. The system of claim 1, wherein the mesoscale flow system comprises a flow channel extending from the inlet port and in fluid communication with a reaction chamber.

9. The system of claim 8, wherein the reaction chamber has a larger cross-sectional dimension than the flow channel.

10. The system of claim 8, wherein the reaction chamber has a cross-sectional width of 1-5 mm.

11. The system of claim 9, wherein the reaction chamber has a cross-sectional depth of 2-50 μ

m.

12. The system of claim 1, wherein the sensor is capable of detecting changes in optical properties of fluid within the flow system.

13. The system of claim 12, wherein the sensor is a spectrophotometer.

14. The system of claim 12, wherein the sensor comprises a video camera.

15. The system of claim 1, wherein the sensor is a pressure detector.

16. The system of claim 1, wherein the sensor is an electrical conductivity sensor.

17. The system of claim 1, wherein the appliance comprises a magnetic source.

18. The system of claim 1, wherein the appliance comprises electrical contacts in the nesting site, the device comprises electrical contacts, and the electrical contacts in the appliance mate with electrical contacts on the device when the device is placed in the nesting site.

19. The system of claim 18, wherein the appliance is configured to transmit electrical energy to the device.

20. The system of claim 18, wherein the appliance is configured to receive electrical signals from the device.

21. The system of claim 1, wherein the nesting site comprises a heating element.

22. The system of claim 1, wherein the nesting site comprises a cooling element.

23. The system of claim 1, wherein the device comprises multiple inlet ports.

24. The system of claim 8, wherein the means for thermally cycling fluid comprises a heating/cooling element within the reaction chamber.

25. The system of claim 19, wherein the device comprises a solid substrate, and wherein the means for thermally cycling fluid comprises an electrical heating element integrated into the substrate.

26. The system of claim 1, wherein the sensor is a thermocouple.