MICROFABRICATION METHODS FOR THE OPTIMAL PATTERNING OF SUBSTRATES
First Claim
1. A method of fabricating a microarray, comprising:
- (a) providing a substrate having at least two layers of different chemical reactivity, wherein a well in an outer layer exposes an inner layer;
(b) contacting said substrate with a first reagent specifically reactive with said outer layer to produce a first modified layer;
(c) contacting said substrate with a second reagent specifically reactive with said inner layer of said substrate to produce a modified inner layer, wherein said modified inner layer has a higher affinity for a biopolymer than said modified outer layer, and(d) depositing said biopolymer onto said modified inner layer within said well, wherein said higher affinity of said modified inner layer facilitates localization of said biopolymer onto said well.
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Abstract
The invention is directed to a method of fabricating a microarray. The method includes: (a) providing a substrate having at least two layers of different chemical reactivity, wherein a well in an outer layer exposes an inner layer; (b) contacting the substrate with a first reagent specifically reactive with the outer layer to produce a first modified layer; (c) contacting the substrate with a second reagent specifically reactive with the inner layer of the substrate to produce a modified inner layer, wherein the modified inner layer has a higher affinity for a biopolymer than the modified outer layer, and (d) depositing the biopolymer onto the modified inner layer within the well, wherein the higher affinity of the modified inner layer facilitates localization of the biopolymer onto the well. Methods of fabricating a microarray which include polishing a substrate or functionalizing a plurality of features with a reactive reagent also are provided. A method of fabricating a microarray which includes loading a plurality of discrete nanochannels is additionally provided.
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Citations
23 Claims
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1. A method of fabricating a microarray, comprising:
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(a) providing a substrate having at least two layers of different chemical reactivity, wherein a well in an outer layer exposes an inner layer; (b) contacting said substrate with a first reagent specifically reactive with said outer layer to produce a first modified layer; (c) contacting said substrate with a second reagent specifically reactive with said inner layer of said substrate to produce a modified inner layer, wherein said modified inner layer has a higher affinity for a biopolymer than said modified outer layer, and (d) depositing said biopolymer onto said modified inner layer within said well, wherein said higher affinity of said modified inner layer facilitates localization of said biopolymer onto said well. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method of fabricating a microarray, comprising:
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(a) contacting a substrate having wells with a reagent reactive with said substrate to produce a surface modification within said wells and a surface modification surrounding said wells; (b) polishing said substrate to produce a polished surface surrounding said wells , whereby said surface modification surrounding said wells is removed and said surface modification within said wells is retained, and (c) depositing a biopolymer onto said substrate, wherein higher affinity of said surface modification within said wells compared to said polished surface facilitates localization of said biopolymer within said wells. - View Dependent Claims (9, 10, 11, 12, 13, 14, 15)
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16. A method of fabricating a microarray, comprising:
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(a) functionalizing a plurality of features on a substrate to create discrete single biopolymer anchor sites, said functionalization comprising; (1) contacting said substrate with a reagent reactive with said substrate to produce a modified substrate; (2) applying a protecting reagent to discrete sites on said modified substrate, said discrete sites having an area of between about 5-40 nm2; (3) modifying said reagent located in unprotected regions of said modified substrate surrounding said discrete sites, thereby rendering said regions unreactive to a target biopolymer, and (4) removing said protecting reagent to produce a substrate having a plurality of discrete functionalized features, and (b) attaching a single target biopolymer to one or more of said discrete functionalized features. - View Dependent Claims (17, 18, 19, 20)
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21. A method of fabricating a microarray, comprising:
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(a) contacting a substrate having a plurality of discrete nanochannels with a plurality of biopolymers, said nanochannels having a length and diameter sufficient for entry of only a single biopolymer molecule; (b) applying an electric potential to said substrate sufficient to translocate said single biopolymer molecules into said nanochannels to produce a substrate containing a plurality of single biopolymer molecules each in said plurality of discrete nanochannels, and (c) transferring said plurality of single biopolymer molecules contained in said plurality of discrete nanochannels to a solid support. - View Dependent Claims (22, 23)
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Specification