Methods for genotyping by hybridization analysis
First Claim
1. A method of genotyping, comprising the steps of:
- (a) generating a first diversity panel from nucleic acid molecules of two or more organisms, wherein the first diversity panel comprises a reproducible pattern of nucleic acid molecules;
(b) separating the nucleic acid molecules of the first diversity panel on the basis of sequence or molecular weight;
(c) placing the separated nucleic acid molecules into an addressable array;
(d) generating a second diversity panel from nucleic acid molecules of an organism, wherein the second diversity panel comprises a reproducible pattern of nucleic acid molecules;
(e) hybridizing the second diversity panel to the addressable array; and
(f) detecting hybridization, therefrom determining a pattern of hybridization;
wherein the genotype of the second organism is determined from the hybridization pattern.
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Abstract
This invention provides methods for determining the genotype of organisms by hybridization analysis and, more specifically, to establishing the relatedness of individual organisms within a species. The present invention provides addressable arrays, comprising diversity panels of nucleic acid molecules, in which the molecules on the array are addressable or uniquely identifiable in some fashion. A diversity panel is the result of a method that can distinguish sequence differences between nucleic acid samples. As taught herein, a variety of methods may be used to generate diversity panels. Subsequent to the generation of the diversity panel, the nucleic acid products of the diversity panel are separated for application onto an array. The separated diversity panel is then delivered onto a substrate to create an addressable array and hybridized with labeled nucleic acids. The genotype of an organism is determined by the pattern of hybridization.
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Citations
22 Claims
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1. A method of genotyping, comprising the steps of:
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(a) generating a first diversity panel from nucleic acid molecules of two or more organisms, wherein the first diversity panel comprises a reproducible pattern of nucleic acid molecules;
(b) separating the nucleic acid molecules of the first diversity panel on the basis of sequence or molecular weight;
(c) placing the separated nucleic acid molecules into an addressable array;
(d) generating a second diversity panel from nucleic acid molecules of an organism, wherein the second diversity panel comprises a reproducible pattern of nucleic acid molecules;
(e) hybridizing the second diversity panel to the addressable array; and
(f) detecting hybridization, therefrom determining a pattern of hybridization;
wherein the genotype of the second organism is determined from the hybridization pattern. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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Specification