Solid supports for nucleic acid hybridization assays
First Claim
1. A composition comprising an oligonucleotide covalently attached to amine-containing polymer-coated solid support wherein the covalent attachment occurs between the amine of the polymer-coated solid support and an amine which is tethered to the oligonucleotide.
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Abstract
Compositions and methods for covalently immobilizing an oligonucleotide onto a polymer-coated solid support or similar structure are provided. Specifically, the polymer-coated support, such as a bead, possesses a large number of activatable moieties, preferably primary and secondary amines. An oligonucleotide is activated with a monofunctional or multifunctional reagent, preferably the homotrifunctional reagent cyanuric chloride. The resultant covalently immobilized oligonucleotides on the support serve as nucleic acid probes, and hybridization assays can be conducted wherein specific target nucleic acids are detected in complex biological samples. The beads or similar structures can be employed free in solution, such as in a microtiter well format; in a flow-through format, such as in a column; or in a dipstick. Additionally, dichlorotriazine oligonucleotides and processes for activating oligonucleotides by treatment with cyanuric chloride and derivatives are included in the present invention.
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Citations
19 Claims
- 1. A composition comprising an oligonucleotide covalently attached to amine-containing polymer-coated solid support wherein the covalent attachment occurs between the amine of the polymer-coated solid support and an amine which is tethered to the oligonucleotide.
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11. A method for producing an oligonucleotide covalently linked to a amine-containing polymer, through an amine which is tethered to the 5'"'"' end or the 3'"'"' end or at a location between the 5'"'"' end and 3'"'"' end of the oligonucleotide, which method comprises:
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activating an alkylamine-containing oligonucleotide with cyanuric chloride or a derivative thereof, and conjugating the activated oligonucleotide to an amine-containing polymer. - View Dependent Claims (12, 13, 14, 15, 16)
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17. A method for preparing a support for use in oligonucleotide synthesis, comprising
activating a control pore glass, silica gel or nylon bead support with 3-aminopropyltriethoxysilane; -
reacting the activated support with cyanuric chloride or derivatives thereof; and conjugating the cyanuric chloride reacted support to an amine-containing polymer of poly(ethyleneimine), polyvinylamine, polyallylamine, or poly(p-aminostyrene).
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Specification