Chip, preparation method therefor, and application thereof

1 . A chip, comprising:

a substrate having a surface grafted with an amino group, the amino group being a primary amino group or a secondary amino group; and

a first compound grafted to the surface via the amino group, the first compound comprising the following structure:

wherein R 1 and R 2 are each independently selected from —OH and C1-C5 alkoxy, and n is 1 or 2, a polymer linked to the first compound, wherein the polymer comprises a plurality of structural units, at least one of the structural units comprising an amino group; and

a nucleic acid sequence linked to the polymer, the polymer and the nucleic acid sequence are linked via a linker group, wherein the linker group comprises a first terminus capable of bonding to the nucleic acid sequence and a second terminus capable of linking to the amino group of the polymer, the second terminus comprises at least one of —NHS, an epoxy group and an isocyanate group, and the nucleic acid sequence has a -DBCO or —N 3 modification, and the first terminus comprises -DBCO or —N 3 .

2 . The chip according to claim 1 , wherein R 1 and R 2 are each independently selected from —OH, —OCH 3 and —OCH 2 CH 3 .

3 . The chip according to claim 1 , wherein the first compound comprises one of the following structures.

4 . The chip according to claim 1 , wherein the amino group is derived from at least one of aminosilane, polylysine, polyornithine, chitosan, polyamidoamine dendrimer, polyacrylamide and polyethyleneimine compounds.

5 . The chip according to claim 4 , wherein the amino group is derived from the aminosilane,

optionally wherein the aminosilane is selected from at least one of (3-aminopropyl)triethoxysilane, 3-aminopropyl trimethoxysilane, 3-aminopropyl methyldiethoxysilane, and aminoethylaminopropyl triethoxysilane.

6 . The chip according to claim 1 , wherein the substrate having the surface grafted with the amino group comprises silane linked to the surface and a polymer linked to the surface via the silane, wherein

the silane is selected from at least one of epoxy silane, alkenylsilane and alkynylsilane, and

the polymer comprises a plurality of structural units, at least one of the structural units comprising the amino group,

optionally wherein the polymer is selected from at least one of polylysine, polyornithine, chitosan, polyamidoamine dendrimer, polyacrylamide, and polyethyleneimine compounds.

7 . The chip according to claim 6 , wherein the silane comprises:

(a) the epoxy silane selected from at least one of 3-glycidoxypropyltrimethoxysilane, 3-glycidoxypropyltriethoxysilane, 3-glycidoxypropylmethyldiethoxysilane, and 3-glycidoxypropylmethyldimethoxysilane;

(b) the alkenylsilane selected from at least one of vinyltrimethoxysilane, vinyltriethoxysilane, vinyltris(2-methoxyethoxy) silane, and vinyltriisopropoxysilane; or

(c) the alkynylsilane selected from at least one of propynyloxytrimethoxysilane and 2-propynyl [3-(triethoxysilyl) propyl]carbamate.

8 . The chip according to claim 1 ,

wherein the polymer is selected from at least one of polylysine, polyornithine, chitosan, polyamidoamine dendrimer, polyacrylamide and polyethyleneimine compounds.

9 . A method for preparing a chip, comprising:

obtaining a substrate having a surface grafted with an amino group, the amino group being a primary amino group or a secondary amino group;

linking a first compound to the surface via the amino group, the first compound comprising the following structure:

wherein R 1 and R 2 are each independently selected from —OH and C1-C5 alkoxy, and n is 1 or 2;

optionally wherein R 1 and R 2 are each independently selected from —OH, —OCH 3 and —OCH 2 CH 3 ;

linking a polymer to the first compound, wherein the polymer comprises a plurality of structural units, at least one of the structural units comprising an amino group; and

linking a nucleic acid sequence to the polymer via a linker group, wherein the linker group comprises a first terminus capable of linking to the nucleic acid sequence and a second terminus, preferably the second terminus comprises at least one of an —NHS group, an epoxy group and an isocyanate group, preferably the nucleic acid sequence has a -DBCO or —N 3 modification, and the first terminus comprises -DBCO or —N 3 .

10 . The method according to claim 9 , wherein a reaction is performed at room temperature for 3-5 h to allow the amino group to be linked to the first compound.

11 . The method according to claim 9 , wherein the amino group is derived from an aminosilane,

optionally wherein a reaction is performed at room temperature for 1-8 h to allow the aminosilane to be linked to the surface, in order to obtain the substrate with the surface grafting amino group;

optionally wherein the aminosilane is selected from at least one of (3-aminopropyl)triethoxysilane, 3-aminopropyl trimethoxysilane, 3-aminopropyl methyldiethoxysilane and aminoethylaminopropyl triethoxysilane.

12 . The method according to claim 9 , wherein the substrate having the surface grafted with the amino group comprises silane linked to the surface and a polymer linked to the surface via the silane, wherein

the silane is selected from at least one of epoxy silane, alkenylsilane and alkynylsilane, and

the polymer comprises a plurality of structural units, at least one of the structural units comprising the amino group;

(a) wherein a reaction is performed at 30-65° C. for 1-48 h to allow the polymer to be linked to the silane; optionally

(b) wherein a reaction is performed at room temperature for 1-8 h to allow the silane to be linked to the surface.

13 . The method according to claim 12 , wherein the silane comprises:

(a) the epoxy silane selected from at least one of 3-glycidoxypropyltrimethoxysilane, 3-glycidoxypropyltriethoxysilane, 3-glycidoxypropylmethyldiethoxysilane and 3-glycidoxypropylmethyldimethoxysilane;

(b) the alkenylsilane selected from at least one of vinyltrimethoxysilane, vinyltriethoxysilane, vinyltris(2-methoxyethoxy) silane, and vinyltriisopropoxysilane; or

(c) the alkynylsilane selected from at least one of propynyloxytrimethoxysilane and 2-propynyl [3-(triethoxysilyl) propyl]carbamate.

14 . The method according to claim 9 ,

wherein a reaction is performed at 30-80° C. for 1-48 h to allow the nucleic acid sequence to be linked to the first compound, optionally the method further comprising:

treating the surface with a blocking reagent selected from at least one of NHS-PEG4, NHS-PEG4-NHS, NHS-PEG-N3 and acetic anhydride.

15 . The method according to claim 9 , further comprising:

treating the surface with a blocking reagent selected from at least one of NHS-PEG4, NHS-PEG4-NHS, NHS-PEG-N3 and acetic anhydride.

16 . The method according to claim 15 ,

(a) wherein a reaction is performed at 30-65° C. for 1-48 h to allow the first terminus of the linker group to be linked to the nucleic acid sequence;

(b) wherein a reaction is performed at room temperature for 0.5-5 h to allow the second terminus of the linker group to be linked to the amino group of the polymer;

(c) wherein the polymer is selected from at least one of polylysine, polyornithine, chitosan, polyamidoamine dendrimer, polyacrylamide and polyethyleneimine compounds, wherein a reaction is performed at 30-40° C. for 1-5 h to allow the polymer to be linked to the first compound; or

any combination of (a), (b), and (c).

17 . A method for detecting nucleic acid, comprising a step of contacting the chip of claim 1 and detecting the nucleic acid.

18 . The method according to claim 17 , comprising sequencing the nucleic acid.

19 . The method according to claim 9 , wherein the first compound is selected from one of the following compounds: