Method of sequencing a nucleic acid
First Claim
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1. A method for preparing an array for analysis of a mixture of nucleic acids, the method comprising:
- providing a substrate comprising a cavitated fiber optic wafer formed from a fused bundle of a plurality of individual optical fibers, each individual optical fiber having a diameter between 3 and 100 μ
m, the wafer comprising a top surface and a bottom surface, the top surface comprising at least 1,000 wells, wherein said wells are etched into the top surface of the cavitated fiber optic wafer and wherein the thickness of the wafer between the top surface and the bottom surface is between 0.5 mm and 5.0 mm;
wherein the depth of each well ranges from between one half the diameter of an individual optical fiber and three times the diameter of an individual optical fiber;
depositing the nucleic acids on the cavitated fiber optic wafer by attachment to said wells or to beads delivered to said wells; and
delivering a solution comprising a pyrophosphate sequencing reagent to said wells, thereby preparing the array for analysis of said mixture of nucleic acids.
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Abstract
Disclosed herein are methods and apparatuses for sequencing a nucleic acid. These methods permit a very large number of independent sequencing reactions to be arrayed in parallel, permitting simultaneous sequencing of a very large number (>10,000) of different oligonucleotides.
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Citations
19 Claims
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1. A method for preparing an array for analysis of a mixture of nucleic acids, the method comprising:
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providing a substrate comprising a cavitated fiber optic wafer formed from a fused bundle of a plurality of individual optical fibers, each individual optical fiber having a diameter between 3 and 100 μ
m, the wafer comprising a top surface and a bottom surface, the top surface comprising at least 1,000 wells, wherein said wells are etched into the top surface of the cavitated fiber optic wafer and wherein the thickness of the wafer between the top surface and the bottom surface is between 0.5 mm and 5.0 mm;
wherein the depth of each well ranges from between one half the diameter of an individual optical fiber and three times the diameter of an individual optical fiber;depositing the nucleic acids on the cavitated fiber optic wafer by attachment to said wells or to beads delivered to said wells; and delivering a solution comprising a pyrophosphate sequencing reagent to said wells, thereby preparing the array for analysis of said mixture of nucleic acids. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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Specification