Nucleic acid sequencing using microsphere arrays
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Abstract
The invention relates to DNA sequencing by synthesis techniques, including those utilizing the detection of pyrophosphate (PPi) generated during the DNA synthesis reaction (pyrosequencing). The methods and compositions utilize biosensor arrays comprising microspheres distributed on a surface.
315 Citations
45 Claims
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1-21. -21. (canceled)
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22. A method of sequencing a plurality of different target nucleic acids, comprising:
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(a) providing a population of microspheres distributed at discrete sites on a surface of a substrate, wherein each discrete site comprises a microsphere attached to a hybridization complex comprising a different target nucleic acid and a primer, and wherein one or more enzymes used to generate a signal from pyrophosphate is attached at said discrete sites (b) extending said primer at each of said discrete sites, thereby adding nucleotides complementary to said target nucleic acid and releasing pyrophosphate; and
(c) detecting said pyrophosphate using said one or more enzymes, thereby sequencing said plurality of target nucleic acids. - View Dependent Claims (23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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Specification