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Method for modifying silicon dioxide material by DNA, product of method and application of product

A technology of silica and nano-silica, applied in the field of materials, can solve problems such as interactions that are rarely explored

Active Publication Date: 2022-01-28
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, metal ion-mediated DNA assembly on the surface of nanomaterials is still poorly studied
Most of the developed methods focus on the adsorption of long DNA (e.g. genetic DNA, plasmids, etc.)

Method used

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  • Method for modifying silicon dioxide material by DNA, product of method and application of product
  • Method for modifying silicon dioxide material by DNA, product of method and application of product
  • Method for modifying silicon dioxide material by DNA, product of method and application of product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: SiNPs synthesis and characterization

[0022] SiNPs according to conventional prepared by method. The solution containing absolute ethanol, ammonia (28-30%), and deionized water was stirred for 5 minutes to ensure complete mixing. Then add tetraethyl orthosilicate absolute ethanol solution to the above solution, and react at room temperature for 24 hours. The synthesized SiNPs were stored in ethanol solution at room temperature.

[0023] Before use, SiNPs were collected by centrifugation at 12000 rpm for 10 min, washed three times with Milli-Q water, and then redispersed in Milli-Q water for subsequent experiments. The size distribution and zeta potential of SiNPs were measured using a Malvern Zetasizer Nano ZS system, and their morphology was characterized using transmission electron microscopy (TEM).

[0024] The results of transmission electron microscopy (TEM) and dynamic light scattering (DLS) indicated that spherical SiNPs with uniform size (diamet...

Embodiment 2

[0025] Example 2: Screening of metal ions mediating DNA modification on the surface of SiNPs

[0026] (1) DNA modified SiNPs method

[0027] 10nM FAM / Cy3-labeled DNA (pH7.4, 10mM HEPES), 100μg mL -1 SiNPs (pH7.4, 10mM HEPES) were mixed with different concentrations of metal ions (pH7.4, 10mM HEPES), incubated at room temperature for 5 minutes, and washed with buffer (10mM HEPES, 1mM metal ions, pH7.4) after centrifugation. Granules 3 times. The fluorescence spectrum of the adsorbed DNA (Ex=490nm, Em=518nm) was recorded using a fluorescence spectrometer.

[0028] (2) Metal ion screening

[0029] The fluorophore 6-carboxyfluorescein (FAM) was labeled on the 5′ end of the 25-mer DNA polyadenosine chain (FAM-A25), and the metal ions that could effectively mediate the surface assembly of DNA and SiNPs were screened. Just mix FAM-A25, SiNPs and metal ions, and get a uniform green solution under UV light (365nm) irradiation. After centrifugation, the SiNPs precipitated and show...

Embodiment 3

[0038] Embodiment 3: DNA microarray construction

[0039] Use the SpotBot 2 microarray spotter to dissolve the solution in the spotting buffer (10mM Tris-HCl, 100μM ZrCl 4 , pH 7.4) were printed onto clean glass slides. The average diameter of the sample spots is 100 μm, and the distance between two spots is 50 μm. Prepared slides were incubated for 5 minutes at room temperature in a humidified chamber. Subsequently, wash twice with Tris-HCl buffer (10 mM Tris-HCl, pH 7.4), and immerse the slide in blocking solution (Tris-HCl buffer containing 10 μM poly15A, pH 7.4) at room temperature for 5 minutes to blunt surface. After washing twice with Tris-HCl buffer, the slides were stored at 4°C for further use.

[0040] DNA microarrays have been widely used in in vitro DNA analysis and clinical diagnosis. In traditional methods, the assembly of DNA probes on the surface of glass slides mainly relies on silane chemistry, which is complicated and time-consuming. Given Zr 4+ Does...

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Abstract

The invention discloses a method for modifying a silicon dioxide material through DNA, a product of the method and application of the product, and belongs to the technical field of materials. According to the method disclosed by the invention, the short-chain DNA is simply, quickly and stably fixed on the surface of the silicon dioxide material by using metal ion-assisted DNA, and through comprehensive screening of the metal ions, some metal ions are found to be capable of effectively mediating modification of short-chain DNA molecules on the surface of the silicon dioxide material; and the DNA modification density can be adjusted by changing the concentration of DNA or the metal ions. The method for preparing the DNA modified silicon dioxide material and the prepared product can be applied to DNA microarrays and biological sensing and have wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of materials, and more specifically relates to a method for modifying a silicon dioxide material with DNA, a product thereof and an application of the product. Background technique [0002] Silica nanoparticles (SiNPs), with their size tunability and high biocompatibility, are one of the most attractive nanomaterials in life sciences. Given that DNA molecules can realize various biological applications such as molecular assembly, biocatalysis, biosensing, gene regulation, drug delivery, etc., the functionalization of SiNPs with oligonucleotide DNA molecules can effectively improve their utilization. [0003] Generally, the as-synthesized SiNPs are inert to DNA molecules because both of them are highly hydrophilic and negatively charged, making it difficult to directly immobilize DNA strands on the surface of SiNPs. Currently, there are mainly two approaches to modify DNA strands on the surface of SiNPs. On...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6837
CPCC12Q1/6806C12Q1/6837C12Q2563/137C12Q2563/155C12Q2565/507
Inventor 黄悦颜玉婷
Owner NANJING FORESTRY UNIV
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