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 forming an array of polymers on a surface of a substrate, comprising:
providing a substrate comprising a solid support having a top surface and a bottom surface;
disposing on said top surface a dichroic antireflective coating having an outermost surface;
derivatizing said outermost surface to provide reactive functional groups protected by a photolabile protecting groups;
photolyzing, with a wavelength of light that is photolytic to said photolabile protecting groups, a first selected region to remove said photolabile protecting groups in said first selected region and to provide exposed functional groups in said first selected region;
coupling monomers protected by photolabile protecting groups, to said exposed functional groups in said first selected region; and
repeating said steps of photolyzing and coupling in one or more subsequent selected regions overlapping all or part of said first selected region to form a plurality of different and positionally distinct polymer sequences contained in different features,
wherein said antireflective coating absorbs greater than about 75% of said wavelength of light that is photolytic to said photolabile protecting groups.
2. The method according to claim 1 , wherein said array of polymers is an array of oligonucleotides.
3. The method according to claim 1 , wherein said outermost surface further comprises SiO 2 to make said antireflective coating suitable for chemical functionalization.
4. The method according to claim 1 , wherein said reactive functional groups are selected from the group consisting of hydroxyl, carboxyl, amino, thiol, haloalkyl, 1,2-diol, aldehyde, acryloyl, maleimidyl and N-succinimidylcarboxylate groups.
5. The method according to claim 1 , wherein said features are smaller than about 20 μm.
6. The method according to claim 5 , wherein said features are smaller than about 10 μm.
7. The method according to claim 1 , further comprising derivatizing said outermost surface with a silanation reagent.
8. The method according to claim 7 , wherein said silanation reagent is selected from the group consisting of Bis-hydroxyethyl-3-aminopropyltriethoxysilane, N(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine and Bis(trimethoxysilyl)-ethane.
9. The method according to claim 7 , wherein said silanation reagent is N(2-hydroxyethyl)-N,N-bis(trimethoxysilylpropyl)amine.
10. The method according to claim 7 , wherein said silanation reagent is a mixture of Bis-hydroxyethyl-3-aminopropyltriethoxysilane and Bis(trimethoxysilyl)-ethane.
11. The method according to claim 1 , wherein said photolabile protecting group is MeNPOC.
12. The method according to claim 2 , wherein said functionalization is with a silanation reagent.
13. The method of claim 1 , wherein said antireflective coating absorbs less than about 40% of visible light and reflects less than about 10% of said wavelength of light that is photolytic to said photolabile protecting groups.
14. The method of claim 1 , wherein said wavelength of light that is photolytic to said photolabile protecting groups is in the UV range.
15. The method of claim 14 , wherein said wavelength of light that is photolytic to said photolabile protecting groups has a wavelength above 340 nanometers.
16. The method of claim 15 , wherein said wavelength of light that is photolytic to said photolabile protecting groups has a wavelength above 365 nanometers.