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Solid-phase substrate as well as preparation method and application thereof

一种基底、基团的技术，应用在固相基底及其制备领域，能够解决影响测序质量、稳定性差、核酸分子分布密度不易控制等问题，达到提高芯片质量、改善性能、改善测序质量的效果

Pending Publication Date: 2022-05-24
GENEMIND BIOSCIENCES CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the actual production and R&D process, there will be problems such as difficult control of the distribution density of nucleic acid molecules and poor stability, which directly affect the quality of sequencing

Method used

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  • Solid-phase substrate as well as preparation method and application thereof
  • Solid-phase substrate as well as preparation method and application thereof
  • Solid-phase substrate as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0089] Example 1 Place at 57°C after silanization

[0090] (1) Cleaning the glass substrate: clean the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0091] (2) Activated glass substrate: soaked in piranha solution for 30 minutes, washed with ultrapure water for 5 times, and washed with ethanol once (surface hydroxylation);

[0092] (3) Surface silane derivatization reaction: prepare 2% (mass concentration) GOPTS (3-(2,3-glycidoxy)propyltrimethoxysilane) ethanol solution, react at 37°C for 5 hours, and then purify 5 times Wash with water, 5 times of ethanol, 1 time of acetone, dry with nitrogen, and set aside;

[0093] (4) Low temperature placement treatment: place the prepared chips in a nitrogen drying cabinet, placement conditions: temperature 57°C, relative humidity 55%, placement time 4 days;

[0094] (5) Amino-DNA-CY3 molecule immobilization: prepare 0.25M phosphate buffer containing amino-DNA-CY3 molecules, coat the surface, react at 37°C for 10...

Embodiment 2

[0096] Example 2 Place at 47°C after silanization

[0097] (1) Cleaning the glass substrate: clean the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0098] (2) Activated glass substrate: soaked in piranha solution for 30 minutes, washed with ultrapure water for 5 times, and washed with ethanol once;

[0099] (3) Surface silane derivatization reaction: prepare 2% (mass concentration) GOPTS ethanol solution, react at 37° C. for 5 hours, then clean with 5 times of pure water, 5 times of ethanol and 1 time of acetone, dry with nitrogen, and set aside for later use;

[0100] (4) Low temperature placement treatment: place the prepared chips in a nitrogen drying cabinet, placement conditions: temperature 47°C, relative humidity 55%, placement time 4 days;

[0101] (5) Amino-DNA-CY3 molecule immobilization: prepare 0.25M phosphate buffer containing amino-DNA-CY3 molecules, coat the surface, react at 37°C for 10 hours, and then cross-wash with 1XPBS (pH=7.4) an...

Embodiment 3

[0103] Example 3 Place at 37°C after silanization

[0104] (1) Cleaning the glass substrate: clean the surface with 5% (mass concentration) hydrochloric acid for 3 hours;

[0105] (2) Activated glass substrate: soaked in piranha solution for 30 minutes, washed with ultrapure water for 5 times, and washed with ethanol once;

[0106] (3) Surface silane derivatization reaction: prepare 2% (mass concentration) GOPTS ethanol solution, react at 37° C. for 5 hours, then clean with 5 times of pure water, 5 times of ethanol and 1 time of acetone, dry with nitrogen, and set aside for later use;

[0107] (4) Low temperature placement treatment: place the prepared chips in a nitrogen drying cabinet, placing conditions: temperature 37°C, relative humidity 55%, placement time 4 days;

[0108] (5) Amino-DNA-CY3 molecule immobilization: prepare 0.25M phosphate buffer containing amino-DNA-CY3 molecules, coat the surface, react at 37°C for 10 hours, and then cross-wash with 1XPBS (pH=7.4) and ...

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Abstract

The invention relates to the technical field of nucleic acid chips, in particular to a solid-phase substrate and a treatment method and application thereof. The solid-phase substrate is provided with at least one silanized surface, the silanized surface is a second surface formed by exposing a silanized first surface to a specific environment for a certain time, the specific environment is an inert gas environment, the temperature of the specific environment is selected from 37 DEG C to 60 DEG C, and the certain time is not less than 2 hours. The performance of the solid-phase substrate is improved, so that the distribution density and the stability of nucleic acid molecules fixed on the surface of the solid-phase substrate are improved, and the quality of the nucleic acid chip is finally improved.

Description

technical field [0001] The present invention relates to the technical field of nucleic acid chips, in particular to a solid-phase substrate and a preparation method and application thereof. Background technique [0002] Chips are an important tool in nucleic acid sequencing and are widely used in various types of nucleic acid sequencing, especially the third-generation single-molecule sequencing chips are randomly immobilized nucleic acid (such as DNA) fragments on glass and other substrates to form millions of single molecules. obtained by molecular points. The density and stability of nucleic acid molecules are mainly determined by the density and distribution of the active group coating on the substrate surface. [0003] In the prior art, silane molecules are usually used to perform a silylation reaction on the surface of a chip substrate to form a silylated modified surface, that is, an active group coating. The chip substrate after the silanization reaction is general...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C03C17/30C12Q1/6874
CPCC12Q1/6874C03C17/30C03C2217/70C03C2218/11C03C2218/31
Inventor 赵智陆永艺王琦颜钦赵陆洋
Owner GENEMIND BIOSCIENCES CO LTD
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