Methods and systems for sequencing long nucleic acids
First Claim
Patent Images
1. A method for sequencing a target nucleic acid, comprising:
- removing the target nucleic acid from a partial copy of the target nucleic acid hybridized to the target nucleic acid, wherein the partial copy of the target nucleic acid comprises;
(i) a capture probe from a plurality of capture probes;
(ii) a first solution probe from a plurality of first solution probes;
(iii) a second solution probe from a plurality of second solution probes; and
(iv) a third solution probe from a plurality of third solution probes; and
sequencing one or more bases of the partial copy of the target nucleic acid by extending a primer hybridized to a 3′
end of the partial copy of the target nucleic acid to generate a first extension product, thereby obtaining a first sequence read to determine the sequence of the partial copy of the target nucleic acid by performing a sequencing reaction, wherein the target nucleic acid is from a plurality of target nucleic acid fragments; and
wherein the capture probe is ligated to the first solution probe, the first solution probe is ligated to both the capture probe and the second solution probe, the second solution probe is ligated to both the first and third solution probes, and the third solution probe comprises a universal primer sequence at the 3′
end complementary to the primer.
2 Assignments
0 Petitions
Accused Products
Abstract
The present invention provides methods and systems for sequencing long nucleic acid fragment. The present invention also provides a method of sequencing a target polynucleotide with fewer probes. Further, 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.
133 Citations
14 Claims
-
1. A method for sequencing a target nucleic acid, comprising:
-
removing the target nucleic acid from a partial copy of the target nucleic acid hybridized to the target nucleic acid, wherein the partial copy of the target nucleic acid comprises; (i) a capture probe from a plurality of capture probes; (ii) a first solution probe from a plurality of first solution probes; (iii) a second solution probe from a plurality of second solution probes; and (iv) a third solution probe from a plurality of third solution probes; and sequencing one or more bases of the partial copy of the target nucleic acid by extending a primer hybridized to a 3′
end of the partial copy of the target nucleic acid to generate a first extension product, thereby obtaining a first sequence read to determine the sequence of the partial copy of the target nucleic acid by performing a sequencing reaction, wherein the target nucleic acid is from a plurality of target nucleic acid fragments; and
wherein the capture probe is ligated to the first solution probe, the first solution probe is ligated to both the capture probe and the second solution probe, the second solution probe is ligated to both the first and third solution probes, and the third solution probe comprises a universal primer sequence at the 3′
end complementary to the primer. - View Dependent Claims (2, 3, 4, 9, 10, 11, 12, 13, 14)
-
-
5. The method of 4, prior to said ligating the capture probe and the first solution probe, further comprising:
hybridizing the target nucleic acid to the capture probe and the first solution probe, wherein the capture probe comprises sequence complementary to a first region of a known sequence of the target nucleic acid, wherein the first solution probe is in solution,- and wherein said hybridizing is configured to permit ligation of the capture probe and the first solution probe. - View Dependent Claims (6, 7, 8)
Specification