Genetic analysis by sequence-specific sorting
First Claim
1. A method of sorting polynucleotides having predetermined sequence characteristics, the method comprising the steps of:
- extending a primer annealed polynucleotides having predetermined sequence characteristics to incorporate a predetermined terminator having a capture moiety;
capturing polynucleotides having extended primers by a capture agent that specifically binds to the capture moiety; and
melting the captured polynucleotides from the extended primers.
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Accused Products
Abstract
The invention provides methods for sorting polynucleotides from a population based on predetermined sequence characteristics. In one aspect, the method of the invention is carried out by extending a primer annealed polynucleotides having predetermined sequence characteristics to incorporate a predetermined terminator having a capture moiety, capturing polynucleotides having extended primers by a capture agent that specifically binds to the capture moiety, and melting the captured polynucleotides from the extended primers to form a subpopulation of polynucleotides having the predetermined sequence characteristics. In another aspect, the method of the invention is carried out on a population of tagged polynucleotides so that after a subpopulation is selected, the members of the subpopulation may be simultaneously analyzed using the unique tags on the polynucleotides to convey analytical information to a hybridization array for a readout.
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Citations
9 Claims
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1. A method of sorting polynucleotides having predetermined sequence characteristics, the method comprising the steps of:
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extending a primer annealed polynucleotides having predetermined sequence characteristics to incorporate a predetermined terminator having a capture moiety;
capturing polynucleotides having extended primers by a capture agent that specifically binds to the capture moiety; and
melting the captured polynucleotides from the extended primers.
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2. A method of producing a subpopulation of polynucleotides having a complexity less than that of a parent population, the method comprising the steps of:
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annealing a primer to polynucleotides of a parent population to form primer-polynucleotide duplexes;
extending the primer to incorporate a predetermined terminator having a capture moiety;
separating the primer-polynucleotide duplexes having an extended primer from the parent population by specifically binding the capture moiety of the predetermined terminator to a capture agent attached to a solid phase support;
melting the primer-polynucleotide duplexes to form a subpopulation of polynucleotides having a complexity less than that of the parent population.
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3. A method of producing a population of polynucleotides having a desired complexity less than that of a parent population, the method comprising the steps of:
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(a) annealing a primer to polynucleotides of a parent population to form primer-polynucleotide duplexes;
(b) extending the primer to incorporate a predetermined terminator having a capture moiety;
(c) separating the primer-polynucleotide duplexes having an extended primer from the parent population by specifically binding the capture moiety of the predetermined terminator to a capture agent attached to a solid phase support;
(d) melting the primer-polynucleotide duplexes to form a selected population of polynucleotides having a complexity less than that of the parent population, the selected population forming a parent population for subsequent steps;
(e) repeating steps (a) through (d) until a selected population of the desired complexity is obtained. - View Dependent Claims (4, 5, 6, 7, 8)
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9-10. -10. (canceled)
Specification