Microfluidic sequencing systems
First Claim
Patent Images
1. A sequencing apparatus comprising:
- a body having a top portion, a bottom portion and an interior portion;
the interior portion comprising at least two intersecting channels, wherein at least one of the two intersecting channels has at least one cross sectional dimension between about 0.1 μ
m and 500 μ
m;
an electrokinetic fluid direction system for moving a first sequencing reagent through at least one of the two intersecting channels;
a source of sequencing primers;
an electropipettor for introducing sequencing primers from the source of sequencing primers to the at least two intersecting channels;
a mixing zone fluidly connected to the at least two intersecting channels for mixing the first sequencing reagent with a second sequencing reagent;
a size separation zone fluidly connected to the mixing zone for separating sequencing products by size, thereby providing the sequence of a target nucleic acid; and
a computer having computer-readable code for selecting a sequencing primer.
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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.
288 Citations
78 Claims
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1. A sequencing apparatus comprising:
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a body having a top portion, a bottom portion and an interior portion; the interior portion comprising at least two intersecting channels, wherein at least one of the two intersecting channels has at least one cross sectional dimension between about 0.1 μ
m and 500 μ
m;an electrokinetic fluid direction system for moving a first sequencing reagent through at least one of the two intersecting channels; a source of sequencing primers; an electropipettor for introducing sequencing primers from the source of sequencing primers to the at least two intersecting channels; a mixing zone fluidly connected to the at least two intersecting channels for mixing the first sequencing reagent with a second sequencing reagent; a size separation zone fluidly connected to the mixing zone for separating sequencing products by size, thereby providing the sequence of a target nucleic acid; and a computer having computer-readable code for selecting a sequencing primer. - View Dependent Claims (2, 3, 4)
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5. A nucleic acid sequencing system, comprising:
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a body structure comprising a plurality of microfluidic channels, wherein the body structure is fluidly coupled to a pipettor capillary that extends outward from the body structure; a source of one or more template nucleic acid, which template nucleic acid source is capable of being fluidly coupled to at least a first one of the microfluidic channels; a source of one or more sequencing reagent, which sequencing reagent source is capable of being fluidly coupled to the at least first microfluidic channel; a detector; and
,a fluid flow controller that flows the template nucleic acid and the one or more sequencing reagent into contact in the at least first microfluidic channel, whereby one or more sequencing product is formed, wherein the detector detects the one or more sequencing product, thereby determining at least a subsequence of the template nucleic acid. - View Dependent Claims (8, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 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, 63, 64, 65, 66, 67, 68, 69, 70, 74, 75, 76, 77, 78)
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6. A nucleic acid sequencing system, comprising:
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a body structure comprising a plurality of microfluidic channels; a source of one or more template nucleic acid, which template nucleic acid source is capable of being fluidly coupled to at least a first one of the microfluidic channels; a source of one or more sequencing reagent, which sequencing reagent source is capable of being fluidly coupled to the at least first microfluidic channel;
wherein the source of one or more sequencing reagent comprises a set of sequencing primers, the set of primers comprising at least about 70% of all possible primers of a given length;a detector; and
,a fluid flow controller that flows the template nucleic acid and the one or more sequencing reagent into contact in the at least first microfluidic channel, whereby one or more sequencing product is formed, wherein the detector detects the one or more sequencing product, thereby determining at least a subsequence of the template nucleic acid. - View Dependent Claims (9, 16, 17, 62)
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7. A nucleic acid sequencing system, comprising:
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a body structure comprising a plurality of microfluidic channels; a source of one or more template nucleic acid, which template nucleic acid source is capable of being fluidly coupled to at least a first one of the microfluidic channels; a source of one or more sequencing reagent, which sequencing reagent source is capable of being fluidly coupled to the at least first microfluidic channel; a detector; and a fluid flow controller that flows the template nucleic acid and the one or more sequencing reagent into contact in the at least flint microfluidic channel, whereby one or more sequencing product is formed, wherein the detector detects the one or more sequencing product, thereby determining at least a subsequence of the template nucleic acid, wherein the system comprises a sequencing product separation channel into which the one or more sequencing product is flowed and separated by size, or charge, or size and charge, into a plurality of separated sequencing products. - View Dependent Claims (71, 72, 73)
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Specification