Methods of identifying homologous genes using FISH
First Claim
1. A fluorescence in situ hybridization method of distinguishing a first gene in a maternal chromosome from a second gene in a paternal chromosome by single nucleotide polymorphisms which distinguish the first gene from the second gene and wherein the first gene and the second gene are homologs comprisingidentifying a first nucleotide type that is a single nucleotide polymorphism within the first gene,hybridizing a first primer type directly upstream of the first nucleotide type, extending the first primer type across the first nucleotide type in the presence of a first polymerase, first extension nucleotides and a first labeled extension nucleotide complementary to the first nucleotide type, wherein the first labeled extension nucleotide hybridizes to the first nucleotide type,identifying a second nucleotide type that is a single nucleotide polymorphism within the second gene and which is different from the first nucleotide type,hybridizing a second primer type directly upstream of the second nucleotide type, extending the second primer type across the second nucleotide type in the presence of a second polymerase, second extension nucleotides and a second labeled extension nucleotide complementary to the second nucleotide type wherein the second labeled extension nucleotide hybridizes to the second nucleotide type, wherein the first gene is differentially labeled from the second gene.
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Abstract
The present invention relates to methods of hybridizing nucleic acid probes to genomic DNA.
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Citations
20 Claims
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1. A fluorescence in situ hybridization method of distinguishing a first gene in a maternal chromosome from a second gene in a paternal chromosome by single nucleotide polymorphisms which distinguish the first gene from the second gene and wherein the first gene and the second gene are homologs comprising
identifying a first nucleotide type that is a single nucleotide polymorphism within the first gene, hybridizing a first primer type directly upstream of the first nucleotide type, extending the first primer type across the first nucleotide type in the presence of a first polymerase, first extension nucleotides and a first labeled extension nucleotide complementary to the first nucleotide type, wherein the first labeled extension nucleotide hybridizes to the first nucleotide type, identifying a second nucleotide type that is a single nucleotide polymorphism within the second gene and which is different from the first nucleotide type, hybridizing a second primer type directly upstream of the second nucleotide type, extending the second primer type across the second nucleotide type in the presence of a second polymerase, second extension nucleotides and a second labeled extension nucleotide complementary to the second nucleotide type wherein the second labeled extension nucleotide hybridizes to the second nucleotide type, wherein the first gene is differentially labeled from the second gene.
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18. A fluorescence in situ hybridization method of distinguishing a first gene in a maternal chromosome from a second gene in a paternal chromosome by single nucleotide polymorphisms which distinguish the first gene from the second gene and wherein the first gene and the second gene are homologs comprising
identifying a first nucleotide type that is a single nucleotide polymorphism within a first sequence within the first gene, hybridizing a first labeled complementary sequence to the first sequence and with a first labeled nucleotide of the first labeled complementary sequence hybridizing to the first nucleotide type, hybridizing a first complementary sequence to the first sequence and adjacent to the first labeled complementary sequence, ligating the first labeled complementary sequence to the first sequence, identifying a second nucleotide type that is a single nucleotide polymorphism within a second sequence within the second gene, hybridizing a second labeled complementary sequence to the second sequence and with a second labeled nucleotide of the second labeled complementary sequence hybridizing to the second nucleotide type, hybridizing a second complementary sequence to the second sequence and adjacent to the second labeled complementary sequence, ligating the second labeled complementary sequence to the second sequence, wherein the first gene is differentially labeled from the second gene.
Specification