Methods of amplifying and sequencing nucleic acids
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Abstract
An apparatus and method for performing rapid DNA sequencing, such as genomic sequencing, is provided herein. The method includes the steps of preparing a sample DNA for genomic sequencing, amplifying the prepared DNA in a representative manner, and performing multiple sequencing reaction on the amplified DNA with only one primer hybridization step.
106 Citations
96 Claims
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1-84. -84. (canceled)
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85. A method for sequencing nucleic acids comprising:
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(a) fragmenting large template nucleic acid molecules to generate a plurality of fragmented nucleic acids; (b) delivering the fragmented nucleic acids into aqueous microreactors in a water-in-oil emulsion such that a plurality of aqueous microreactors comprise a single copy of a fragmented nucleic acid, a single bead capable of binding to the fragmented nucleic acid, and amplification reaction solution containing reagents necessary to perform nucleic acid amplification; (c) amplifying the fragmented nucleic acids in the microreactors to form amplified copies of said nucleic acids and binding the amplified copies to beads in the microreactors; (d) delivering the beads to areas on a planar surface to produce an array of at least 10,000 reaction sites; and (e) performing a sequencing reaction simultaneously on the nucleic acids at the sites. - View Dependent Claims (86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96)
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Specification