Mesoscale polynucleotide amplification analysis
First Claim
1. A device for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction, the device comprising:
- a solid substrate microfabricated to define;
a sample inlet port; and
a mesoscale flow system comprising;
a sample flow channel extending from said inlet port; and
a polynucleotide polymerization reaction chamber, in fluid communication with said flow channel, containing reagents for the polymerization reaction; and
means for thermally regulating the contents of said chamber whereby the temperature is controlled to amplify said preselected polynucleotide.
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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.
178 Citations
43 Claims
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1. A device for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction, the device comprising:
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a solid substrate microfabricated to define;
a sample inlet port; and
a mesoscale flow system comprising;
a sample flow channel extending from said inlet port; and
a polynucleotide polymerization reaction chamber, in fluid communication with said flow channel, containing reagents for the polymerization reaction; and
means for thermally regulating the contents of said chamber whereby the temperature is controlled to amplify said preselected polynucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. A device for amplifying a preselected polynucleotide in a sample by conducting a polymerase chain reaction (PCR), the device comprising:
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a solid substrate microfabricated to define;
a sample inlet port; and
a mesoscale flow system comprising;
a sample flow channel extending from said inlet port; and
a PCR chamber, in fluid communication with said flow channel, for receiving said preselected polynucleotide and PCR reagents; and
means for thermally cycling the contents of said chamber whereby, in each cycle, temperature is controlled to separate double stranded polynucleotide, and to synthesize polynucleotide, thereby to amplify said preselected polynucleotide. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32)
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33. A method for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction, the method comprising:
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(i) providing a device comprising;
a solid substrate microfabricated to define;
a sample inlet port; and
a mesoscale flow system comprising;
a sample flow channel extending from said inlet port; and
a polynucleotide polymerization reaction chamber in fluid communication with said flow channel; and
means for thermally regulating the contents of said chamber at a temperature controlled to amplify said preselected polynucleotide;
(ii) delivering, through said inlet port and said mesoscale flow system to said reaction chamber, said sample polynucleotide and reagents required for the polymerization reaction; and
(iii) thermally controlling the contents of said chamber to polymerize said polynucleotide. - View Dependent Claims (34, 35, 36, 37, 38, 39, 40, 41, 42, 43)
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Specification