Antireflective coatings for high-resolution photolithographic synthesis of DNA arrays
View Patent ↗The present invention provides an array of polymers and methods of forming arrays of polymers by providing a substrate having a first layer including one or more dielectric coatings on a solid support and a second layer including a plurality of polymers disposed on the first layer. The invention also provides methods for forming an array of polymers on a substrate using light-directed synthesis by providing a substrate having a first layer including one or more dielectric coatings on a solid support, derivatizing the first layer by contacting the first layer with a silanation reagent, and a second layer disposed on said first layer wherein the second layer includes functional groups protected with a photolabile protecting group.
1. A method of synthesizing an array of polymers, the method comprising:
(a) providing a substrate, wherein a surface of the substrate comprises functional groups protected by photolabile protecting groups;
(b) deprotecting the functional groups within selected regions of the surface, wherein deprotecting comprises illumination of the selected regions with light that is photolytic to the photolabile protecting groups, and wherein the selected regions correspond to locations on the surface where features of the array of polymers are synthesized;
(c) coupling monomers to the functional groups within the selected regions, wherein the monomers are protected by photolabile protecting groups; and
(d) repeating steps (b)-(c) for one or more repetitions to synthesize the array of polymers, wherein steps (b)-(c) occur within a synthesis system, wherein the synthesis system comprises a light source and an anti-reflective coating, and wherein the anti-reflective coating reduces reflected illumination within the synthesis system such that the features of the array of polymers are smaller than 1 μm.
2. The method of claim 1 , wherein the size of the features is smaller than 0.5 μm.
3. The method of claim 1 , wherein the anti-reflective coating reflects less than about 10% of the light that is photolytic to the photolabile protecting groups.
4. The method of claim 3 , wherein the anti-reflective coating reflects less than about 5% of the light that is photolytic to the photolabile protecting groups.
5. The method of claim 1 , wherein the anti-reflective coating additionally possesses light absorbing properties.
6. The method of claim 1 , wherein the anti-reflective coating has a transmittance of less than 5% for the light with a wavelength of 365 nm.
7. The method of claim 1 , wherein the light includes one or more wavelengths longer than 340 nm.
8. The method of claim 1 , wherein the light includes a wavelength of 365 nm.
9. The method of claim 1 , wherein the synthesis system additionally comprises a flow cell.
10. The method of claim 9 , wherein the synthesis system additionally comprises a fluid delivery system in fluid communication with the flow cell, wherein the fluid delivery system comprises a plurality of reagent reservoirs, and wherein the plurality of reagent reservoirs contain solutions with monomers for the array of polymers.
11. The method of claim 1 , wherein at least one of the one or more repetitions utilizes a set of selected regions which is different from a previous set of selected regions.
12. The method of claim 1 , wherein light including a wavelength of 365 nm is photolytic to the photolabile protecting groups.
13. The method of claim 12 , wherein the photolabile protecting groups are MeNPOC.
14. The method of claim 1 , wherein the array of polymers comprises polynucleotides.
15. The method of claim 1 , wherein the array of polymers comprises polypeptides.
16. The method of claim 1 , wherein the array of polymers comprises 1,000,000 different features.
17. The method of claim 1 , wherein the substrate possesses two or more arrays of polymers.
18. The method of claim 1 , wherein the substrate is about 1 inch by about 3 inches.
19. The method of claim 1 , wherein the array of polymers occupies a portion of the substrate smaller than about 5 cm by about 5 cm.
20. A method of synthesizing an array of polymers, the method comprising:
(a) providing a substrate, wherein a surface of the substrate comprises functional groups protected by photolabile protecting groups;
(b) deprotecting the functional groups within selected regions of the surface, wherein deprotecting comprises illumination of the selected regions with light that is photolytic to the photolabile protecting groups, and wherein the selected regions correspond to locations on the surface where features of the array of polymers are synthesized;
(c) coupling monomers to the functional groups within the selected regions, wherein the monomers are protected by photolabile protecting groups; and
(d) repeating steps (b)-(c) for one or more repetitions to synthesize the array of polymers, wherein steps (b)-(c) occur within a synthesis system, wherein the synthesis system comprises a light source and an anti-reflective coating, and wherein the anti-reflective coating reduces reflected illumination within the synthesis system such that the features of the array of polymers are smaller than 1 μm, wherein the anti-reflective coating absorbs greater than about 75% of the light that is photolytic to the photolabile protecting groups.
21. The method of claim 20 , wherein the size of the features is smaller than 0.5 μm.
22. The method of claim 20 , wherein the anti-reflective coating reflects less than about 10% of the light that is photolytic to the photolabile protecting groups.
23. The method of claim 20 , wherein the anti-reflective coating reflects less than about 5% of the light that is photolytic to the photolabile protecting groups.
24. The method of claim 20 , wherein the synthesis system additionally comprises a flow cell.
25. The method of claim 24 , wherein the synthesis system additionally comprises a fluid delivery system in fluid communication with the flow cell, wherein the fluid delivery system comprises a plurality of reagent reservoirs, and wherein the plurality of reagent reservoirs contain solutions with monomers for the array of polymers.
26. The method of claim 20 , wherein at least one of the one or more repetitions utilizes a set of selected regions which is different from a previous set of selected regions.
27. The method of claim 20 , wherein light including a wavelength of 365 nm is photolytic to the photolabile protecting groups.
28. The method of claim 27 , wherein the photolabile protecting groups are MeNPOC.
29. The method of claim 20 , wherein the array of polymers comprises polynucleotides.
30. The method of claim 20 , wherein the array of polymers comprises polypeptides.
31. The method of claim 20 , wherein the array of polymers comprises 1,000,000 different features.
32. A method of synthesizing an array of polymers, the method comprising:
(a) providing a substrate, wherein a surface of the substrate comprises functional groups protected by photolabile protecting groups;
(b) deprotecting the functional groups within selected regions of the surface, wherein deprotecting comprises illumination of the selected regions with light that is photolytic to the photolabile protecting groups, and wherein the selected regions correspond to locations on the surface where features of the array of polymers are synthesized;
(c) coupling monomers to the functional groups within the selected regions, wherein the monomers are protected by photolabile protecting groups; and
(d) repeating steps (b)-(c) for one or more repetitions to synthesize the array of polymers, wherein steps (b)-(c) occur within a synthesis system, wherein the synthesis system comprises a light source and an anti-reflective coating, and wherein the anti-reflective coating reduces reflected illumination within the synthesis system such that the features of the array of polymers are smaller than 0.5 μm.
33. A method of synthesizing an array of polymers, the method comprising:
(a) providing a synthesis system and a substrate, wherein the synthesis system comprises a light source, a flow cell, a fluid delivery system in fluid communication with the flow cell, and an anti-reflective coating, wherein the substrate and the flow cell form a flow cell cavity, and wherein a surface of the substrate comprises functional groups protected by photolabile protecting groups;
(b) deprotecting the functional groups within selected regions of the surface, wherein deprotecting comprises illumination of the selected regions with light that is photolytic to the photolabile protecting groups, wherein the light includes at least one wavelength above 340 nm, and wherein the selected regions correspond to locations on the surface where features of the array of polymers are synthesized;
(c) coupling monomers to the functional groups within the selected regions, wherein the monomers are delivered to the flow cell cavity by the fluid delivery system, and wherein the monomers are protected by photolabile protecting groups; and
(d) repeating steps (b)-(c) for one or more repetitions to synthesize the array of polymers, wherein the anti-reflective coating reduces reflected illumination from the light source within the synthesis system such that the features of the array of polymers are smaller than 1 μm.