Silane mixtures
Silanation compositions containing a mixture of two or more silanation reagents, where at least one silanation reagent includes a functional group capable of supporting polymer synthesis and at least one silanation reagent includes no functional group capable of supporting polymer synthesis are useful in modulating the active site density and hydrolytic stability of a surface. These compositions are particularly useful in silanating a surface prior to preparation of a polymer array and provide for increased hybridization results.
1. A method of preparing an array of monomers or polymers comprising:
providing a substrate including a silica surface or other silicon oxide-containing surface thereon;
contacting the silica or the other silicon oxide-containing surface of the substrate with a silanation composition comprising a mixture of a first compound and a second compound such that the first and the second compounds are covalently attached to the surface to form a silanation coating;
the first compound comprising a trialkoxy silyl group and a functional group, wherein said functional group is capable of supporting polymer synthesis and is selected from the group consisting of hydroxyl, amino and carboxyl; the first compound being selected from the group consisting of N-(3-(triethoxysilyl)-propyl)-4-hydroxybutyrarnide; N,N-bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane; 3-((2 hydroxyethyl)methylamino)propyl-trimethoxysilane; N-(2-hydroxy-ethyl)-N,N-bis(trimethoxy-silylpropyl)-amine; 1,2-bis((2-hydroxyethyl)-(3 trimethoxysilylpropyl)aminoethane; 3-acetoxy-propyltrimethoxysilane; (trimethoxysilyloxy)-(C3-C12-alkyl)-(triethoxysilane); and 3-glycidoxypropyltriethoxysilane; and
the second compound comprising a trialkoxy silyl group and wherein the second compound in the mixture does not include the functional group of the first compound; the second compound being selected from the group consisting of 1,2-bis(trimethoxysilyl)-ethane; 1,3-bis(trimethoxysilyl)propane; 1,6-bis(trimethoxysilyl)hexane; 3-(N,N-dimethyl-amino)-propyl-trimethoxysilane; and N,N-bis(3-trimethoxysilylpropyl)-methylamine;
wherein the molar ratio of the first compound to the second compound in the mixture ranges from about 1:50 to 1:500;
and wherein the mixture of the first and second compounds in the silanation coating imparts increased hydrolytic stability to the coating;
protecting the functional group capable of supporting polymer synthesis of the first compound with a protecting group or reacting the functional group capable of supporting polymer synthesis of the first compound with a compound having a reactive group protected by a protecting group to provide the first compound of the silanation coating protected with the protecting group;
removing the protecting group of the first compound in selected regions of the silanation coating to provide an exposed functional or reactive group;
reacting said exposed functional or reactive group with the monomer or the polymer whereby the monomer or the polymer is coupled to the exposed group; and
repeating the steps of removing and reacting to produce the array of monomers or polymers.
2. A method of preparing an array of polymers according to claim 1 , wherein the protecting group is a photoremovable protecting group.
3. A method of preparing an array of polymers according to claim 1 , wherein the step of removing comprises activating a photoacid generator in the selected regions by selective application of light having a predetermined wavelength to provide an acid and exposing the protecting group to the acid.
4. A method of preparing an array of polymers according to claim 1 wherein the first compound is N-(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine and the second compound is 1,2-bis(trimethoxysilyl)ethane.
5. The method of claim 1 , wherein the first compound is N-(3-(triethoxysilyl)propyl)-4-hydroxybutyramide.
6. The method of claim 1 , wherein the first compound is N,N-bis(2-hydroxyethyl)-3aminopropyltriethoxysilane.
7. The method of claim 1 , wherein the first compound is 3-((2-hydroxyethyl)methylamino)propyltrimethoxysilane.
8. The method of claim 1 , wherein the first compound is N-(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine.
9. The method of claim 1 , wherein the first compound is 1,2-bis((2-hydroxyethyl)(3-trimethoxysilylpropyl)aminoethane.
10. The method of claim 1 , wherein the first compound is 3-acetoxypropyltrimethoxysilane.
11. The method of claim 1 , wherein the first compound is (trimethoxysilyloxy)-(C3-C12-alkyl)-(triethoxysilane).
12. The method of claim 1 , wherein the first compound is 3-glycidoxypropyltriethoxysilane.
13. The method of claim 1 , wherein the second compound is 1,2-bis(trimethoxysilyl)ethane.
14. The method of claim 1 , wherein the second compound is bis(trimethoxysilyl)propane.
15. The method of claim 1 , wherein the second compound is 1,6-bis(trimethoxysilyl) hexane.
16. The method of claim 1 , wherein the second compound is 3-(N,N-dimethylamino)propyltrimethoxysilane.
17. The method of claim 1 , wherein the second compound is N,N-bis(3-trimethoxysilylpropyl)-methylamine.
18. The method of claim 1 , wherein the polymers are nucleic acids.
19. The method of claim 1 , wherein the functional group of the first compound capable of supporting polymer synthesis is protected with a protecting group before the polymers of the array are attached to the silanated substrate.
20. The method of claim 1 , wherein the protecting group is an acid labile protecting group.
21. The method of claim 1 , wherein the protecting group is removed by activation of a photoacid generator in selected regions of the silica surface by selective application of light having a predetermined wavelength to provide an acid and exposure of the protecting group to the acid.
22. The method of claim 1 , wherein the first compound is N-(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine and the second compound is 1,2-bis(trimethoxysilyl)ethane.
23. The method of claim 1 , wherein the molar ratio of the second compound to the first compound in the mixture ranges from about 50:1 to 100:1.
24. The method of claim 1 , wherein the molar ratio of the second compound to the first compound in the mixture is about 99:1.
25. The method of claim 1 , wherein the second compound is 1,2-bis(trimethoxysilyl)ethane and the first compound is bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane.
26. The method of claim 1 , wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane in the mixture ranges from about 50:1 to 100:1.
27. The method of claim 1 , wherein the molar ratio of 1,2-bis(trimethoxysilyl)ethane to bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane in the mixture is about 99:1.
28. The method of claim 1 , wherein the silanation coating does not comprise a mixture of bis(2-hydroxyethyl)-3-aminopropyltriethoxysilane and 1,2-bis(trimethoxy silyl)ethane.
29. The method of claim 1 , wherein the silanation coating does not contain 1,2-bis(trimethoxysilyl)ethane.
30. The method of claim 1 , further comprising a hexaethylene glycol linker extending from the functional group capable of supporting polymer synthesis to the polymers of the array.