METHODS OF TARGETED SEQUENCING
First Claim
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1. A method of sequencing comprising:
- (a) forming a hybridization complex comprising;
(i) a first set of capture probes;
(ii) a target polynucleotide; and
(iii) a first solution probe,wherein at least one of the first set of capture probes binds a first region of the target polynucleotide, the first solution probe binds a second region of the target polynucleotide, the first solution probe has a base at the 3′
end complementary to a first loci site of the target polynucleotide, therefore when hybridization occurs said at least one of the first set of capture probes and the first solution probe can be ligated, wherein said first solution probe comprises a cleavable first cap;
(b) ligating said at least one of the first set of capture probes and said first solution probe;
(c) adding a second cap to capture probes of said first set of capture probes not ligated to said first solution probe;
(d) removing said first cap; and
(e) sequencing said target polynucleotide.
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Abstract
The present invention provides methods for targeted sequencing of polynucleotide. In one aspect, the present invention provides a method of sequencing a target polynucleotide with fewer probes. In another aspect, the present invention provides a method of sequencing a target polynucleotide with longer reads. Locus-specific, ligation-assisted sequencing/genotyping method and ligation-captured sequencing method are also provided in the present invention. The methods of the present invention allow low-cost, high-throughput and accurate sequencing of nucleic acids.
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33 Claims
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1. A method of sequencing comprising:
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(a) forming a hybridization complex comprising; (i) a first set of capture probes; (ii) a target polynucleotide; and (iii) a first solution probe, wherein at least one of the first set of capture probes binds a first region of the target polynucleotide, the first solution probe binds a second region of the target polynucleotide, the first solution probe has a base at the 3′
end complementary to a first loci site of the target polynucleotide, therefore when hybridization occurs said at least one of the first set of capture probes and the first solution probe can be ligated, wherein said first solution probe comprises a cleavable first cap;(b) ligating said at least one of the first set of capture probes and said first solution probe; (c) adding a second cap to capture probes of said first set of capture probes not ligated to said first solution probe; (d) removing said first cap; and (e) sequencing said target polynucleotide. - 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, 28, 29, 30, 31, 32, 33)
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Specification