Microfabricated reactor, process for manufacturing the reactor, and method of amplification
First Claim
1. A device for amplifying a preselected polynucleotide in a sample of cells by conducting a polynucleotide amplification reaction, the device comprising:
- a solid substrate which is microfabricated to define;
a sample inlet port;
a flow system for micro- to picoliter volumes, comprising;
a sample flow channel extending from said inlet port; and
a polynucleotide amplification reaction chamber in fluid communication with said flow channel;
said chamber and said channel being of dissimilar dimension; and
a fluid exit port in fluid communication with said flow system;
an integrated component for inducing cell lysis, andmeans for thermally cycling the contents of said chamber whereby in each cycle the temperature is controlled to dehybridize double stranded polynucleotide and to permit the in- vitro amplification of a preselected polynucleotide.
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Accused Products
Abstract
An integrated microfabricated instrument for manipulation, reaction and detection of microliter to picoliter samples. The instrument is suited for biochemical reactions, particularly DNA-based reactions such as the polymerase chain reaction, that require thermal cycling since the inherently small size of the instrument facilitates rapid cycle times. The integrated nature of the instrument provides accurate, contamination-free processing. The instrument may include reagent reservoirs, agitators and mixers, heaters, pumps, and optical or electromechanical sensors. Ultrasonic Lamb-wave devices may be used as sensors, pumps and agitators.
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Citations
18 Claims
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1. A device for amplifying a preselected polynucleotide in a sample of cells by conducting a polynucleotide amplification reaction, the device comprising:
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a solid substrate which is microfabricated to define; a sample inlet port; a flow system for micro- to picoliter volumes, comprising; a sample flow channel extending from said inlet port; and a polynucleotide amplification reaction chamber in fluid communication with said flow channel;
said chamber and said channel being of dissimilar dimension; anda fluid exit port in fluid communication with said flow system; an integrated component for inducing cell lysis, and means for thermally cycling the contents of said chamber whereby in each cycle the temperature is controlled to dehybridize double stranded polynucleotide and to permit the in- vitro amplification of a preselected polynucleotide. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for amplifying a preselected polynucleotide in a sample of cells by conducting a polynucleotide amplification reaction, the method comprising:
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(i) providing a device comprising;
a solid substrate microfabricated to define;a sample inlet port; a flow system for micro- to picoliter volumes comprising; a sample flow channel extending from said inlet port; and a polynucleotide amplification reaction chamber in fluid communication with said flow channel;
said chamber and said channel being of dissimilar dimension; anda fluid exit port in fluid communication with said flow system; and means for thermally regulating the contents of said chamber at a temperature controlled to permit amplification of said preselected polynucleotide; (ii) delivering, through said inlet port and said flow system to said reaction chamber, a sample of cells and reagents required for an in vitro polynucleotide amplification reaction; and (iii) thermally controlling the contents of said chamber to permit amplification of said polynucleotide. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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Specification