Modified surfaces
Provided herein are methods and compositions for coating surfaces with polymers. The methods and compositions are suited for conducting biological reactions.
1. A method for performing an enzymatic reaction, comprising:
(a) providing a substrate comprising surface-bonded initiator species comprising a silane;
(b) conducting surface initiated polymerization of a mixture of (i) first monomers comprising acrylamide and/or ethoxylated acrylamide, each of which has no biomolecule attached to, and (ii) second monomers comprising acrydite, each of which has a biomolecule attached to, wherein said surface initiated polymerization starting from said initiator species, and coupling a plurality of said biomolecules to a polymer brush coating during said surface initiated polymerization, thereby producing said polymer brush coating with said plurality of said biomolecules coupled to said polymer brush coating; and
(c) performing one or more enzymatic reactions with said biomolecules on said substrate; wherein the biomolecules are selected from the group consisting of: oligonucleotides and polynucleotides, wherein the one enzymatic reaction is selected from the group consisting of: polymerase chain reaction, sequencing reaction, sequencing by synthesis reaction, ligation reaction, extension reaction, and transcription reaction.
2. The method of claim 1 , further comprising applying heat to said substrate.
3. The method of claim 1 , wherein said polymer brush coating coupled with said plurality of biomolecules exhibits robustness wherein at least 90% of said biomolecules are retained on said surface after 40 cycles of sequencing by synthesis reactions.
4. The method of claim 1 , wherein the substrate comprises at least 1,000,000 different types of biomolecules, and wherein each biomolecule is an oligonucleotide.
5. The method of claim 4 , wherein said enzymatic reaction is an extension reaction.
6. A method for making a modified surface, comprising:
(a) providing a surface;
(b) covalently bonding initiator species to said surface, wherein said initiator species comprises a silane; and
(c) conducting surface initiated polymerization of a mixture of (i) first monomers comprising acrylamide and/or ethoxylated acrylamide, each of which has no biomolecule attached to, and (ii) second monomers comprising acrydite, each of which has a biomolecule attached, wherein said surface initiated polymerization starting polymer from said initiator species, thereby producing a polymer brush coating comprising a plurality of polymer chains, each of which has said biomolecule attached.
7. The method of claim 6 , wherein said initiator species comprises the molecule shown below:
8. The method of claim 6 , wherein said second monomers are 5′ acrydite modified oligonucleotides.
9. The method of claim 1 , wherein said polymer brush coating coupled with said plurality of biomolecules exhibits robustness in said polymerase chain reactions and wherein said polymerase chain reactions comprise the following reaction conditions:
(a) a denaturation step at a temperature of at least 85° C. for at least 15 seconds;
(b) an annealing step at a temperature of at least 50° C. for at least 15 seconds; and
(c) an extension step at a temperature of at least 70° C. for at least 30 seconds.
10. The method of claim 1 , wherein said first monomers comprise acrylamide.
11. The method of claim 10 , wherein said first monomers further comprise N-(2-hydroxyethyl)acrylamide.
12. The method of claim 1 , wherein said surface-bonded initiator species comprises 2-bromo-2-methyl-N,N-bis-(3-trimethoxysilanylpropyl)propionamide.
13. The method of claim 1 , wherein said surface initiated polymerization comprises atom-transfer radical polymerization (ATRP).
14. The method of claim 1 , wherein said second monomers are 5′ acrydite modified oligonucleotides or polynucleotides.
15. The method of claim 1 , wherein said first monomers comprise ethoxylated acrylamide.
16. The method of claim 6 , wherein said first monomers comprise ethoxylated acrylamide.
17. The method of claim 1 , wherein robustness of said polymer brush coating coupled with said plurality of biomolecules is exhibited in said polymerase chain reaction: at least 90% of said biomolecules are retained on said surface after 25 cycles of said polymerase chain reactions.