Mesoscale polynucleotide amplification device and method
DCFirst 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 having a cross-section and extending from said inlet port; and
a polynucleotide polymerization reaction chamber having a cross-section and being in fluid communication with said flow channel, said chamber containing reagents for the polymerization reaction, said flow channel and said reaction chamber having at least one cross-sectional dimension of width or depth which is between about 0.1 to 500 μ
m said reaction chamber cross-section and said flow channel cross-section being dissimilar;
means for thermally regulating the contents of said chamber whereby the temperature is controlled to amplify said preselected polynucleotide; and
means for detecting said amplified 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.
75 Citations
13 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 having a cross-section and extending from said inlet port; and
a polynucleotide polymerization reaction chamber having a cross-section and being in fluid communication with said flow channel, said chamber containing reagents for the polymerization reaction, said flow channel and said reaction chamber having at least one cross-sectional dimension of width or depth which is between about 0.1 to 500 μ
m said reaction chamber cross-section and said flow channel cross-section being dissimilar;
means for thermally regulating the contents of said chamber whereby the temperature is controlled to amplify said preselected polynucleotide; and
means for detecting said amplified polynucleotide. - View Dependent Claims (5, 6, 7, 8, 9, 10)
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2. 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 having a cross-section and extending from said inlet port; and
a polynucleotide polymerization reaction chamber having a cross-section and being in fluid communication with said flow channel, said flow channel and said reaction chamber having at least one cross-sectional dimension of width or depth which is between about 0.1 to 500 μ
m said reaction chamber cross-section and said flow channel cross-section being dissimilar; 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;
(iii) thermally controlling the contents of said chamber to polymerize said polynucleotide; and
(iv) detecting said amplified polynucleotide. - View Dependent Claims (11, 12, 13)
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3. A device for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction, said device comprising at least one micromachinable solid substrate and including:
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a sample inlet port; and
a mesoscale flow system comprising;
a sample flow channel having a cross-section and extending from said inlet port; and
a polynucleotide polymerization reaction chamber having a cross-section and being in fluid communication with said flow channel, said flow channel and said reaction chamber having at least one cross-sectional dimension of width or depth which is between about 0.1 to 500 μ
m said reaction chamber cross-section and said flow channel cross-section being dissimilar;
means for thermally regulating the contents of said chamber whereby the temperature is controlled to amplify said preselected polynucleotide; and
means for detecting said amplified polynucleotide.
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4. 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 at least one micromachinable solid substrate and including;
a sample inlet port; and
a mesoscale flow system comprising;
a sample flow system comprising;
a sample flow channel having a cross-section and extending from said inlet port; and
a polynucleotide polymerization reaction chamber having a cross-section and being in fluid communication with said flow channel, said flow channel and said reaction chamber having at least one cross-sectional dimension of width or depth which is between about 0.1 to 500 μ
m said reaction chamber cross-section and said flow channel cross-section being dissimilar; 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 for the polymerization reaction;
(iii) thermally controlling the contents of said chamber to polymerize said polynucleotide; and
(iv) detecting said amplified polynucleotide.
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Specification