METHODS AND SYSTEMS FOR SOLUTION BASED SEQUENCE ENRICHMENT
First Claim
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1. A method for isolating and reducing the complexity of a plurality of nucleic acid sequences, the method comprising, in order, the steps of:
- a) providing;
i) a solid support wherein said solid support comprises hybridization probes immobilized on the support and hybridizable to target nucleic acid sequences, ii) a fragmented nucleic acid sample comprising target nucleic acid sequences,b) amplifying said immobilized hybridization probes to produce amplification products that comprise a binding moiety, wherein said amplification products are maintained in solution,c) hybridizing said nucleic acid sample to said amplification products in solution such that hybridization between said amplification products and target nucleic acid sequences is allowed to occur to produce hybridization complexes,d) separating the target nucleic acid/amplification product hybridization complexes from non-specifically hybridized nucleic acids by said binding moiety, ande) eluting the hybridized target nucleic acid sequences from the complexes thereby isolating and reducing the complexity of a plurality of nucleic acid sequences.
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Abstract
The present invention provides methods and systems for the capture and enrichment of target nucleic acids and analysis of the enriched target nucleic acids. In particular, the present invention provides for the enrichment of targeted sequences in a solution based format.
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Citations
30 Claims
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1. A method for isolating and reducing the complexity of a plurality of nucleic acid sequences, the method comprising, in order, the steps of:
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a) providing;
i) a solid support wherein said solid support comprises hybridization probes immobilized on the support and hybridizable to target nucleic acid sequences, ii) a fragmented nucleic acid sample comprising target nucleic acid sequences,b) amplifying said immobilized hybridization probes to produce amplification products that comprise a binding moiety, wherein said amplification products are maintained in solution, c) hybridizing said nucleic acid sample to said amplification products in solution such that hybridization between said amplification products and target nucleic acid sequences is allowed to occur to produce hybridization complexes, d) separating the target nucleic acid/amplification product hybridization complexes from non-specifically hybridized nucleic acids by said binding moiety, and e) eluting the hybridized target nucleic acid sequences from the complexes thereby isolating and reducing the complexity of a plurality of nucleic acid sequences. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 28, 29, 30)
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15. A method for isolating and reducing the complexity of a plurality of nucleic acid sequences the method comprising, in order, the steps of:
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a) providing;
i) a solid support wherein said solid support comprises hybridization probes immobilized on the support and hybridizable to target nucleic acid sequences, ii) a nucleic acid sample comprising target nucleic acid sequences,b) amplifying said immobilized hybridization probes to produce the amplification products that comprise a binding moiety, wherein said amplification products are maintained in solution, c) hybridizing said nucleic acid sample to said amplification products in solution such that hybridization between said amplification products and target nucleic acids is allowed to occur to produce hybridization complexes, d) separating the target nucleic acid/amplification product hybridization complexes from non-specifically hybridized nucleic acids by said binding moiety, e) eluting the hybridized target nucleic acid sequences from the complexes thereby isolating and reducing the complexity of a plurality of nucleic acid sequences, and f) sequencing the eluted target nucleic acid sequences. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27)
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Specification