Functional micro-well chip for trace detection on nucleic acid molecules and preparation method of functional micro-well chip

Abstract

The invention relates to a functional micro-well chip for trace detection on nucleic acid molecules and a preparation method of the functional micro-well chip. The preparation method of the functional micro-well chip comprises the following steps: on the basis of taking nano silicon dioxide as a substrate, modifying the substrate with octadecyltrichlorosilane, covering the substrate with a mask, and degrading the octadecyltrichlorosilane in an uncovered region by use of ultraviolet light, thereby obtaining patterned nano silicon dioxide super-hydrophilic micro-wells; and modifying the region of the super-hydrophilic micro-wells with a functionalizing silane reagent, wherein the functionalizing silane reagent is capable of adding functional groups to the region of the super-hydrophilic micro-wells by use of covalent bonds, and the functional groups can be combined with ligands to form the functional micro-well chip having biological specific affinity. The functional micro-well chip has the advantage of fixed-point confinement enrichment, so that sample application is simple and convenient, sample application points are regular and uniform, and the accuracy of detection is improved; meanwhile, the functional micro-well chip is excellent in optical properties and stable in chemical properties; a nucleic acid capture chain is modified by securely bonding the corresponding ligands with the covalent bonds so that the nucleic acid immobilization amount can be increased, and trace detection on biological nucleic acid molecules can be realized.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, functional materials and detection and analysis, and in particular relates to a functionalized microwell chip for nucleic acid biomolecule trace detection and a preparation method thereof. Background technique [0002] Biochip technology is a new technology developed in the late 1980s, which transforms the chemical process of life into a controllable static form. It arranges a large number of biomacromolecular probes, gene fragments, oligonucleotides, etc. in a specific way on a fixed position on a special carrier, and interacts with the sample to be tested under specific conditions, and records, Detection, analysis. The fastest-growing and most widely used detection-type biochip is nucleic acid chip technology, which utilizes the characteristics of nucleic acid molecules that can be denatured and hybridized, and is inferred by the hybridization of fixed probes or sample nucleic aci...

AI Technical Summary

Problems solved by technology

The method of directly modifying the surface of a smooth substrate is simple, low in cost, and easy to prepare, but the disadvantages are irregular spots, small amount of nucleic acid immobilization, and low signal intensity.

The other is to construct a micro-nano structure on a smooth substrate and then modify it. The micro-nano structure increases the specific surface area and increases the amount of solidified nucleic acid. Inconvenient for signal scanning, detection and analysis

Therefore, this single micro-nano structure cannot achieve fixed-point and controlled detection of nucleic acid molecules. In addition, how to reduce the detection limit and improve sensitivity is still a difficult problem for nucleic acid chip technology.

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