Buffer composition and method for hybridization of microarrays on adsorbed polymer siliceous surfaces
First Claim
1. A method of hybridizing a microarray of oligonucleotides bound to a polymer adsorbed on a surface of a siliceous substrate with a nucleic acid material comprising the step ofincubating the nucleic acid material with the microarray of oligonucleotides on the adsorbed polymer surface in a hybridization solution at a hybridization temperature ranging from about 55°
- C. to about 70°
C. so as to hybridize the nucleic acid material, wherein the hybridization solution comprises a buffer composition that comprises a pH within a range of pH 6.4 to 7.5, a non-chelating buffering agent that maintains the pH within the pH range, and a monovalent cation in a monovalent cation concentration ranging from about 0.01 M to about 2.0 M.
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Accused Products
Abstract
A buffer composition, method and kit for hybridizing microarrays of nucleic acids bound to an adsorbed polymer surface of a siliceous substrate provide an envelope of conditions to hybridize nucleic acid targets, while preserving theintactness of the adsorbed polymer surface of the array. The buffer composition comprises a non-chelating buffering agent, a pH within a range of pH 6.4 and 7.5, a monovalent cation having a monovalent cation concentration that ranges from about 0.01 M to about 2.0 M, and optionally relatively lower concentrations of a chelating agent and an ionic surfactant. The total cation concentration of the buffer composition ranges from about 0.02 M to about 2.0 M. The method comprises incubating the targets with the microarray in the buffer composition at a temperature between about 55° C. and 70° C.
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Citations
20 Claims
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1. A method of hybridizing a microarray of oligonucleotides bound to a polymer adsorbed on a surface of a siliceous substrate with a nucleic acid material comprising the step of
incubating the nucleic acid material with the microarray of oligonucleotides on the adsorbed polymer surface in a hybridization solution at a hybridization temperature ranging from about 55° - C. to about 70°
C. so as to hybridize the nucleic acid material,wherein the hybridization solution comprises a buffer composition that comprises a pH within a range of pH 6.4 to 7.5, a non-chelating buffering agent that maintains the pH within the pH range, and a monovalent cation in a monovalent cation concentration ranging from about 0.01 M to about 2.0 M. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
- C. to about 70°
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17. A method of performing a high temperature hybridization assay comprising the step of:
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incubating a nucleic acid material with a microarray of oligonucleotides in a hybridization solution at a hybridization temperature ranging from about 55°
C. to about 70°
C. so as to hybridize the nucleic acid material,wherein the microarray comprises a siliceous substrate with an adsorbed polymer surface and oligonucleotides bound to the adsorbed polymer surface, and wherein the hybridization solution comprises a pH within a range of pH 6.4 and 7.5 and a buffer composition, the buffer composition comprising a non-chelating buffering agent that maintains the pH within the range and a monovalent cation having a monovalent cation concentration ranging from 0.01 M and 2.0 M.
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18. A method of hybridizing a microarray of oligonucleotides with a nucleic acid material comprising the step of:
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incubating the nucleic acid material with the microarray of oligonucleotides in a hybridization solution at a hybridization temperature ranging from about 55°
C. to about 70°
C. so as to hybridize the nucleic acid material, the oligonucleotides being bound to a polymer coating adsorbed on a surface of a siliceous substrate, the adsorbed polymer coating being non-covalently bound to the siliceous substrate surface,wherein the hybridization solution comprises a buffer composition that comprises a pH within a range of pH 6.4 to 7.5, a non-chelating buffering agent that maintains the pH within the pH range, and a monovalent cation in a monovalent cation concentration ranging from about 0.01 M to about 2.0 M. - View Dependent Claims (19)
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20. A method of reducing surface degradation to a microarray of oligonucleotides during a high temperature hybridization assay comprising:
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incubating a nucleic acid material with the microarray of oligonucleotides in a hybridization solution at a hybridization temperature ranging from about 55°
C. to about 70°
C. so as to hybridize the nucleic acid material, the oligonucleotides being bound to a polycationic polymer that is adsorbed to a surface of a siliceous substrate, the adsorbed polycationic polymer being non-covalently bound to the siliceous substrate surface,wherein the hybridization solution comprises a buffer composition that comprises a pH within a range of pH 6.4 to 7.5, a non-chelating buffering agent that maintains the pH within the pH range, and a monovalent cation in a monovalent cation concentration ranging from about 0.01 M to about 2.0 M.
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Specification