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

A nucleic acid molecule and functionalization technology, applied in biochemical equipment and methods, bioreactors/fermenters for specific purposes, biochemical instruments, etc. Irregular points and other problems, to achieve the effect of pollution-free preparation process, regular sampling points, and lower detection limit

Active Publication Date: 2014-12-10
UNIV OF SCI & TECH BEIJING
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Soak 2×1 square centimeter glass pieces in hot Piranha solution (98% concentrated sulfuric acid / 30% hydrogen peroxide, V / V=7:3) for 1 hour. After cooling, ultrasonically clean with acetone, ethanol, and deionized water for 10 minutes, respectively. Finally, blow it off with nitrogen and dry it in a drying oven; light the candle (avoid using the wax head for new candles, and do not use the wax tail for too short candles). After the flame is stable, use tweezers to pick up the clean glass piece at a constant speed ( 2cm / s) over the stable burning flame and repeatedly translate 15 times, the glass surface will physically deposit a layer of carbon nanoparticles uniformly distributed with a thickness of about 19 microns. Take two small beakers of 5 ml, add 2 ml of tetraethoxysilane to one of them and add 2 ml of ammonia to the other, then put the glass sheet with deposited carbon nanoparticles and the two small beakers into the desiccator respectively, after sealing Put...

Embodiment 2

[0042] (1) Soak 2×1 square centimeter glass pieces in hot Piranha solution (98% concentrated sulfuric acid / 30% hydrogen peroxide, V / V=7:3) for 1 hour. After cooling, ultrasonically clean with acetone, ethanol, and deionized water for 10 minutes, respectively. Finally, blow it off with nitrogen and dry it in a drying oven; light the candle (avoid using the wax head for new candles, and do not use the wax tail for too short candles). After the flame is stable, use tweezers to pick up the clean glass piece at a constant speed ( 2cm / s) over the stable burning flame and repeatedly translate 7 times, the glass surface will physically deposit a layer of uniformly distributed carbon nanoparticles with a thickness of about 10 microns. Take two small beakers of 5 ml, add 2 ml of tetraethoxysilane to one of them and add 2 ml of ammonia to the other, then put the glass sheet with deposited carbon nanoparticles and the two small beakers into the desiccator respectively, after sealing Put ...

Embodiment 3

[0055] (1) Soak a 2×1 square centimeter quartz plate in hot Piranha solution (98% concentrated sulfuric acid / 30% hydrogen peroxide, V / V=7:3) for 1 hour. After cooling, ultrasonically clean with acetone, ethanol, and deionized water for 10 minutes, respectively. Finally, blow it off with nitrogen and dry it in a drying oven; light a candle (avoid using a wax head for a new candle, and do not use a wax tail for a candle that is too short), and after the flame is stable, use tweezers to pick up a clean quartz piece at a constant speed ( 2cm / s) over the stable burning flame and repeatedly translate 9 times, the quartz surface will physically deposit a layer of carbon nanoparticles uniformly distributed with a thickness of about 12 microns. Take two 5ml small beakers, add 2ml of tetraethoxysilane to one of them and add 2ml of ammonia water to the other, then put the quartz sheet with deposited carbon nanoparticles and the two small beakers into the desiccator respectively, after se...

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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...

Claims

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Application Information

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IPC IPC(8): C12M1/00
CPCC12Q1/6834
Inventor 许利苹陈艳霞王树涛
Owner UNIV OF SCI & TECH BEIJING
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