High density electrophoresis device and method
First Claim
1. Apparatus for electrophoretic separation of analytes, the device comprising:
- a substrate including (1) a central reservoir region, (2) a plurality or electrophoretic channels in fluid communication with, and emanating substantially radially from, the central reservoir region, the channels being coplanar with each other, and each channel having (i) a proximal end which is linked to the central reservoir region, and (ii) a distal end, and (3) for each channel one or more chambers that are each linked by a passageway in fluid communication with the distal end of that channel, wherein each passageway leads from each chamber in a direction that is initially away from the central reservoir region, whereby centrifugation of the substrate about a central axis that is perpendicular to the plane of the channels is effective to disperse liquid from the central reservoir region into said channels and chambers such that any air bubbles in the chambers, channels, and passageways are forced towards the axis of rotation, when such liquid is present in the central reservoir region.
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Abstract
The present invention provides, in one aspect, an apparatus for electrophoretic separation of analytes. In one aspect, the apparatus comprises a disc-shaped substrate defining (1) a central reservoir region, (2) a plurality of electrophoretic channels in fluid communication with, and emanating substantially radially from, the central reservoir region, the channels being coplanar with each other, and each channel having (i) a proximal end which is linked to the reservoir region, and (ii) a distal end, and preferably (3) for each channel, at least one chamber, and preferably three chambers, linked by a passageway in fluid communication with the distal end of that channel. Preferably, each passageway leads from each chamber in a direction that is initially away from the central reservoir region, whereby centrifugation of the substrate about a central axis that is perpendicular to the channels is effective to disperse liquid from the central reservoir region into the channels and chambers, such that any air bubbles in the chambers, channels, and passageways are forced towards the axis of rotation, when such liquid is present in the central reservoir region.
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Citations
23 Claims
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1. Apparatus for electrophoretic separation of analytes, the device comprising:
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a substrate including (1) a central reservoir region, (2) a plurality or electrophoretic channels in fluid communication with, and emanating substantially radially from, the central reservoir region, the channels being coplanar with each other, and each channel having (i) a proximal end which is linked to the central reservoir region, and (ii) a distal end, and (3) for each channel one or more chambers that are each linked by a passageway in fluid communication with the distal end of that channel, wherein each passageway leads from each chamber in a direction that is initially away from the central reservoir region, whereby centrifugation of the substrate about a central axis that is perpendicular to the plane of the channels is effective to disperse liquid from the central reservoir region into said channels and chambers such that any air bubbles in the chambers, channels, and passageways are forced towards the axis of rotation, when such liquid is present in the central reservoir region. - 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)
providing an apparatus as defined in claim 1 such that the reservoir region either contains a liquid or is in fluid communication with a liquid source, centrifuging the substrate about a central axis that is perpendicular to the channels so that the liquid is dispersed from the central reservoir region into said channels and chambers, such that any air bubbles in the chambers, channels, and/or passageways are forced towards the axis of rotation, yielding a plurality of bubble-free electrophoretic paths between said reservoir and said chambers. -
17. A method for preparing a plurality of electrophoretic paths which are substantially bubble-free, comprising
providing an apparatus as defined in claim 1 such that the reservoir region, and optionally the channels, passageways, and/or chambers contain a liquid, centrifuging the substrate about a central axis that is perpendicular to the channels so that the liquid is dispersed from the central reservoir region into said channels and chambers, such that any air bubbles in the chambers, channels, and/or passageways are forced towards the axis of rotation, yielding a plurality of bubble-free electrophoretic paths between said reservoir and said chambers. -
18. A method for analyzing a plurality of samples comprising:
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providing an apparatus in accordance with claim 1, such that the central reservoir region, channels, and chambers contain a liquid medium suitable for electrophoresis of such samples, applying an electric field under conditions effective to cause migration of sample through at least one of said channels towards the central reservoir region, and interrogating at least one of the channel(s) to detect one or more sample components in the channel(s).
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19. The method of claim 18, wherein the components to be detected are nucleic acids.
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20. The method of claim 19, wherein the nucleic acids to be detected are fluorescently labeled.
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21. The method of claim 19, wherein the samples are prepared by polymerase chain reaction amplification of one or more selected target sequences.
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22. The method of claim 19, wherein the samples are prepared by ligating at least two oligonucleotides when the oligonucleotides are bound to adjacent regions or a target polynucleotide that are complementary to the at least two oligonucleotides.
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23. The method of claim 18, wherein the components to be detected are polypeptides.
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Specification