Method for isolation of independent, parallel chemical micro-reactions using a porous filter
First Claim
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1. A membrane reactor array comprising:
- a planar array layer comprising a plurality of pores with reagent-carrying beads disposed in a plurality of said pores, with a maximum of one bead per pore, wherein the array is substantially permeable to aqueous solutions but is substantially impermeable to said beads; and
a porous high flow resistance membrane layer, in contact with the planar array layer, that is permeable to nucleic acids and proteins to provide flow resistance of greater than 10 fold that of the flow resistance of the aqueous solutions through a bead-containing pore in said planar array.
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Abstract
The present invention relates to methods and apparati for conducting densely packed, independent chemical reactions in parallel in a substantially two-dimensional array. Accordingly, this invention also focuses on the use of this array for applications such as DNA sequencing, most preferably pyrosequencing, and DNA amplification.
63 Citations
71 Claims
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1. A membrane reactor array comprising:
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a planar array layer comprising a plurality of pores with reagent-carrying beads disposed in a plurality of said pores, with a maximum of one bead per pore, wherein the array is substantially permeable to aqueous solutions but is substantially impermeable to said beads; and
a porous high flow resistance membrane layer, in contact with the planar array layer, that is permeable to nucleic acids and proteins to provide flow resistance of greater than 10 fold that of the flow resistance of the aqueous solutions through a bead-containing pore in said planar array. - 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, 24, 25, 26, 27, 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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28. A membrane reactor array, comprising
(a) a planar array layer comprising a plurality of pores with reagent-carrying beads disposed in a plurality of said pores such that a maximum of one bead is disposed within any one pore, wherein said beads restrict but do not completely block the flow of fluid through said pores, and wherein the array is substantially permeable to aqueous solutions but is substantially impermeable to said beads, and wherein said planar array is made from a fiber material of substantially uniform cross-section weaved to form a mesh layer comprising an array of pores of uniform size, and wherein each pore is an opening of less than 40 μ - m; and
(b) a porous high flow resistance membrane layer, in contact with the planar array layer, that is permeable to nucleic acids and proteins to provide flow resistance of greater than 10 fold that of the flow resistance of aqueous solutes through a bead containing pore in said planar array. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38)
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Specification