Microfluidic systems and methods of genotyping
DCFirst Claim
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1. A method of genotyping a sample material in a microscale device, the method comprising:
- flowing a sample comprising a nucleic acid into a microscale chamber or channel in the microscale device, the microscale chamber or channel having at least one reaction region having a cross sectional dimension of between about 0.1 μ
m and about 500 μ
m;
amplifying a plurality of sequences from the sample in the reaction region of the device to form an amplified sample; and
detecting a plurality of polymorphisms in the amplified sample.
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Abstract
Integrated systems, apparatus, software, and methods for performing biochemical analysis, including DNA sequencing, genomic screening, purification of nucleic acids and other biological components and drug screening are provided. Microfluidic devices, systems and methods for using these devices and systems for performing a wide variety of fluid operations are provided. The devices and systems of are used in performing fluid operations which require a large number of iterative, successive or parallel fluid manipulations, in a microscale, or sealed and readily automated format.
401 Citations
73 Claims
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1. A method of genotyping a sample material in a microscale device, the method comprising:
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flowing a sample comprising a nucleic acid into a microscale chamber or channel in the microscale device, the microscale chamber or channel having at least one reaction region having a cross sectional dimension of between about 0.1 μ
m and about 500 μ
m;
amplifying a plurality of sequences from the sample in the reaction region of the device to form an amplified sample; and
detecting a plurality of polymorphisms in the amplified sample. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 72)
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2. A method of genotyping a sample material in a microfluidic device, the method comprising:
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flowing a sample comprising a nucleic acid into a microscale chamber or channel in the microscale device, the microscale chamber or channel having at least one reaction region with a cross sectional dimension of between about 0.1 μ
m and about 500 μ
m;
modifying the complexity of the sample in the reaction region of the device to form an modified sample; and
,detecting a plurality of polymorphisms in the modified sample. - View Dependent Claims (13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 73)
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27. A system for determining a sequence of nucleotides in a target nucleic acid sequence, comprising:
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a microfluidic device comprising at least a first analysis channel and at least a first probe introduction channel, wherein the analysis channel intersects and is in fluid communication with the probe introduction channel;
a source of the target nucleic acid sequence in fluid communication with the analysis channel;
a plurality of separate sources of oligonucleotide probes in fluid communication with the probe introduction channel, each of the plurality of separate sources containing an oligonucleotide probe having a different nucleotide sequence of length n;
a sampling system for separately transporting a volume of each of the oligonucleotide probes from the sources of oligonucleotide probes to the probe introduction channel and injecting each of the oligonucleotide probes into the analysis channel to contact the target nucleic acid sequence; and
a detection system for identifying whether each oligonucleotide probe hybridizes with the target nucleic acid sequence. - View Dependent Claims (28, 29, 30, 31, 32, 33, 34)
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35. A microfluidic system, comprising:
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a microfluidic device comprising at least two intersecting microscale channels;
a storage element separable from the microfluidic device and comprising a reagent storage substrate and one or more reagents; and
a sampling element operationally coupled to the storage element and at least one of the intersecting microscale channels;
wherein the one or more reagents comprise at least about 70% of all possible oligonucleotides having a length of n nucleotides. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71)
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Specification