Selective attachment of nucleic acid molecules to patterned self-assembled surfaces

A method of attaching pre-formed, hybridizable, synthetic nucleic acid oligomers to selected portions along a surface of a substrate to form a pattern of hybridizable nucleic acid oligomers thereon, comprising the steps of:

• covalently binding a coating of non-DNA-binding organosilane molecules to said surface of said substrate, thus forming an organosilane-coated assembly;
• defining a pattern including a first region, said first region including said non-DNA-binding organosilane molecules, and a second region in which said non-DNA-binding organosilane molecules have been removed from the surface by exposing said organosilane-coated assembly to a pattern of ultraviolet radiation, thereby forming a patterned surface;
• covalently binding nucleic acid-binding organosilane molecules to said patterned surface at said second region, said nucleic acid-binding organosilane molecules having, at a site distal from a site of attachment of said nucleic acid-binding organosilane molecule to said surface of said second region, at least one active site incapable of binding to said surface, incapable of binding to the non-DNA-binding organosilane molecules of said coating, and incapable of binding to said nucleic acid-binding organosilanes, but available as an attachment site for binding a heterobifunctional crosslinker, said heterobifunctional crosslinker including a first reactive site that covalently binds to said at least one active site of said organosilane molecule of said coating, and a second reactive site available for directly and covalently binding to said thiol group of said modified synthetic nucleic acid molecules;
• reacting said organosilane coating with said heterobifunctional crosslinker;
• exposing, after said reacting step, said patterned surface having said nucleic acid-binding organosilanes therein to said modified pre-formed synthetic nucleic acid oligomers to selectively and covalently attach said thiol group of said pre-formed, synthetic nucleic acid oligomers to said second region and to non-specifically adsorb said modified pre-formed nucleic acid oligomers to said patterned surface;
• and
• removing said non-specifically adsorbed pre-formed synthetic nucleic acid oligomers from said patterned surface, thereby forming a pattern of covalently attached hybridizable synthetic nucleic acid oligomers upon said substrate.

A method of attaching pre-formed synthetic hybridizable nucleic acid oligomers, modified to include a thiol group, to selected portions along a surface of a substrate to form a pattern of said modified hybridizable synthetic nucleic acid oligomers thereon, comprising the steps

• covalently binding an organosilane coating to said surface of said substrate, the molecules of said covalently-bound organosilane coating having, at a site distal from a site of attachment of said coating molecule to said surface, at least one active site incapable of binding to said surface and incapable of binding to the other molecules of said coating, the ability of which to covalently bind a heterobifunctional crosslinker is reversed by exposure to irradiation, thus forming an organosilane-coated assembly;
• defining a pattern in said organosilane coating, said pattern including a first region, said first region including said coating molecules, and said at least one active site of said coating molecules in said first region having the ability to bind said heterobifunctional crosslinker, and a second region that lacks coating molecules with the ability to bind said heterobifunctional crosslinker, by exposing said coated assembly to a pattern of radiation thereby forming a patterned surface;
• reacting said organosilane coating with said heterobifunctional crosslinker, said heterobifunctional crosslinker including a first reactive site that covalently binds to said at least one active site of said coating molecules in said first region, and a second reactive site available for directly and covalently binding to said thiol group of said modified synthetic nucleic acid oligomers;
• exposing said patterned surface having said nucleic acid-binding organosilanes therein to said modified pre-formed synthetic nucleic acid oligomers to selectively and covalently bind said thiol group of said modified synthetic nucleic acid oligomers to said first region and to non-specifically adsorb said modified preformed synthetic nucleic acid oligomers to said patterned surface;
• and
• removing said non-specifically adsorbed pre-formed synthetic nucleic acid oligomers from said patterned surface, thereby forming a pattern of covalently attached hybridizable synthetic nucleic acid oligomers upon said substrate.